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USER MANUAL - Loligo Systems

1. 31 Experiment settings 37 Fiber DDC uiia ns ratu a si bii 23 File gt Continue experiment 36 File gt EXIE Lese i ostendam qn 36 File gt New experiment 21 File gt Stop experiment 40 File NaMe ess coeant sme 33 Fish respiration 16 Flow through respirometry 17 altri PECES 38 Flush period ss 34 Flush pump osoei 18 GS E pP RU DD 38 40 Help gt ADOMHE isse etnies 36 40 HAL qo 0 P 26 27 28 TAY ONC ecco 5 35 In 4915 anb 5 35 Hysteresis esee 35 36 Hi qe UL E m 25 26 27 28 TABU DV xitisccanievannicuaimnearaniucs 27 Input Indicatob asus oiov pa 26 Intermittent flow respirometry 18 LEDS 4 List of DEl DS srei m iQ pO Rn Rua lineae 1 LO TMU ssccocasccsibeuesitcbarcetetnenecss feel 25 26 28 Loop number orte rt mne 38 L oii o LOLIGOSYSTEMS COM Loop remaining ert iet cete 38 Measure period 34 Measurement amp Automation 4 13 Metabolites seessssssss 18 DO enha DR 39 MOV AV sas otnvrdo ibi eii 25 26 27 31 NEXE HAS EET 24 29 Note THEN Gs ios oii PE REHRH HER PREHRE 34 Ds solublliby oce cttaeit t rt re Pei 34 Diss rasa cheats abdestut ids cu nud cee eed fees 35 Open respirometry o on 1
2. 5 3 3 3 Measure period s Use this field to specify the duration of the measure periods during the intermittent flow respirometry experiment Please note The field is limited to values between 1 s to 35 weeks 5 3 3 4 O solubility mg O2 l kPa Use this field to specify the oxygen solubility of sample water Please note This field is limited to values between 0 to 0 75 mg L kPa The default value is 0 4855 corresponding to a 15 C fresh water For further information see the appendix page 45 and 46 5 3 3 5 Note field In this field the user can write a note about the experiment The note is saved to the data file Page 34 of 50 L oii o LOLIGOSYSTEMS COM 5 3 4 Chamber field 1 4 5 3 4 1 Resp vol 1 Use this field to specify the volume of water used for the calculation of the MO value Resp vol chamber volume tube volume volume of organism s To specify the Resp vol fill the chamber and the tubes with water and measure the weight on a scale Now empty the chamber and the tubes for water and measure the weight again The difference between the two measurements will be the weight of the chamber volume and the tube volume Multiply the calculated weight with the mass specific volume of water for finding the volume for the chamber and the tubes To find the volume of the organism measure the weight of the organism and multiply with the mass specific volume of the organism Please note This field is l
3. Page 18 of 50 Screen 17 DAQ 4 instrument FLUSH RECIRC O2 N2 TEMP 100 240VAC LOLIGOSYSTEMS COM Flush pump Recirc pump Respirometer Screen 17 shows how to connect the flush and the recirc pump to the respirometer by tubes The connection to the DAQ 4 instrument dotted lines is described above Page 19 of 50 5 Using AutoResp 4 5 1 Startup LOLIGOSYSTEMS COM e Click on the AutoResp 4 icon in the Start menu by following the folder hierarchy shown on Screen 18 Screen 18 Internet Internet Explorer e E mail EJ Microsoft Outlook amp Quattro Track amp LoliTRACK x Microsoft Excel AutoResp 4 9 Measurement amp Autq New Office Document E Newshortcut2 a Open Office Document SetProgram Access and Defaults W Windows Catalog b Windows Update WinZip Accessories Ei Acer ePm e Acer GridVista e AcerSystem Ei AntiVir PersonalEdition Classic e C950 Multi Channel Gas Exchange Software CyberLink PowerDVD Ei Eltima Software e Games Ef LABTECH NOTEBOOKpro Version 12 1 Ef Launch Manager e Measurement Computing Ei Microsoft Office Tools Ef National Instruments i NTI Backup NOW 4 E NTI CD amp DVD Maker 7 E Philips SPC 900NC PC Camera e Philips ToUcam Camera f Pinnade PCTv UsB2 Ei REALTEK Gigabit and Fast Ethernet NIC D
4. amp My System i Bill Data Neighborhood Name Value es El Al amp il Devices and Interfaces Diserial Number Oxo ill NI DAQmx Devices NI NI Devisimu DAQmx Device Wf NI USB 6215 Dev2 Basics amp PXI PXI System Unidentified aA scales a Software E Remote Systems What do you want to do Bunthe N DAQmx TestPanels Remove the device View or change device properties m 8 Y screen 19 bmp Paint XX NLUSB 6215 Devisi Page 42 of 50 L oii o LOLIGOSYSTEMS COM Now change the device name of the NI USB 6215 DAQ card to Dev1 The DAQ 4 instrument should now work properly with AutoResp 4 Screen 29 B NI USB 6215 Dev1 Measurement amp Automation Explorer Jes File Edit View Tools Help Configuration 4 Wf Properties X Dele Eb Self Test ME TestPanels fue ResetDevice Mf Create Task d Configure TEDS Hide Help amp Q My System p Ns m amp Bill Data Neighborhood Name Value Gec gg Al S loce canes seca unie Desear c gii NI DAQmx Devices NI I9 NIUSB 6215 Dev Isimu DAQmx Device V NIUSB 6215 Devi E Basics PXI PXI System Unidentified a Scales a Software E 9 Remote Systems What do you want to do Bun the N DAQmx Test Panels Remove the device View or change device properties Bm Attributes ff Device Routes amp l Calibration
5. A dialogue appears informing you that you will change the last calibration values Click the Continue button to store the new calibration data WARNING This will change last calibration values Cancel 3 Repeat this procedure for all other channels by typing in the respective channel into the Channel Window to record the 100 air saturation calibration value cal 100 4 To record the second calibration value oxygen free water dip the plastic tubing into the vessel containing the cali bration solution 0 cal 0 and fill the syringe slowly with it Please ensure that there are no air bubbles located in the glass tube of the flow through cell Calibration 37 Wait about 3 minutes until the phase angle is constant the variation of the phase angle should be smaller than 0 05 and press the CAL 0 button to store the 0 air sat and temp at 0 values Afterwards press the cali bration solution into the waste A message window opens and informs you that you will change the last calibration values Click the OK button to store the new calibration data Repeat this procedure for all other channels by typing in the respective channel into the Channel Window to record the oxygen free calibration value cal 0 5 Rinse the sensor membranes gently with water after calibration to remove sulfite remaining on the surface 7 2 2 User defined calibration If temperature and phase angle of a former calibration of the same se
6. O to oxygen concentration at time t mg O liter O2 ti oxygen concentration at time t mg O liter V respirometer volume minus volume of experimental animal liter t t tp hour BW body weight of experimental animal kg An advantage of this method is its simplicity A disadvantage is that the measurements are never made at a constant oxygen level due to the continuous use of oxygen by the animal inside the respirometer This might cause problems when interpreting data since animal respiration often changes with ambient oxygen partial pressure Furthermore metabolites from the experimental animal i e CO accumulate in the water thus limiting the duration of measurements This limited time for measurements prevents the experimental animal to recover from initial handling stress that often increase fish respiration significantly and for several hours thus overestimating oxygen consumption rates Page 16 of 50 L oii 0 LOLIGOSYSTEMS COM 4 2 Flow through respirometry or open respirometry This is a more sophisticated method for oxygen consumption measurements Experimental animals are placed in a flow through chamber with known flow rate Oxygen is measured in the inflow and outflow and oxygen consumption rate can be calculated as VO F O2lin O2 ou BW VO oxygen consumption rate mg O kg hour F water flow rate I hour O2 in oxygen content in water inflow mg O liter O2 out
7. 131 35 46 1999 Appendix 55 12 2 Determination of the Oxygen Concentration Using a Modified Stern Volmer Equation The Stern Volmer equation 4 displays a linear correlation between tan o tan or to t and the oxygen concen tration Ob tan t ane et Rsv Oe 4 tan t Do phase angle of oxygen free water o measured phase angle Ksv Stern Volmer constant O2 oxygen content in 96 air saturation The Stern Volmer plots of all our sensors show a distinct non linearity in their response behavior which is also observed for many other oxygen sensors described in literature This non linear response behavior can be described with a modified Stern Volmer equation 5 tan f 1 f 0 1 1 5 tan 1 Ky O 1 Ks 0 This model is based on the assumption that the indicator is distributed in the polymer matrix at two different sites and each fraction fi 1 f1 shows a different quenching constant Ksv Ksvz For practical use this model is not very convenient since it has too many parameters which have to be calibrated Therefore two simplified models based on equation 5 can be used In the first model one fraction of the indicator is assumed to be non quenchable Ksy2 0 tan _ f tan 1 K 0O f 6 The oxygen content in 96 air saturation can be calculated according to equation 7 tani tan tanc Ka 1 f 2 m D 1 O A second model which a
8. 20 132 20 218 20 187 20 097 20 152 20 113 20 068 20 058 20 013 19 964 19 909 19 742 20 019 19 963 19 526 19 796 19 579 19 248 19 117 18 895 19 301 19 169 18 947 97 85 20 367 20 348 20 326 20 302 20 274 20 242 20 207 20 167 20 123 20 073 20 018 19 850 19 633 19 353 19 221 18 999 20 423 20 382 20 357 20 329 20 298 20 262 20 222 20 178 20 128 19 904 19 406 20 478 20 437 20 412 20 384 20 353 20 317 20 277 20 233 20 183 19 958 19 459 98 92 20 589 20 570 20 548 99 19 20 645 20 625 20 603 99 45 20 534 20 700 20 681 20 492 20 467 20 439 20 408 20 372 20 332 20 288 20 237 20 012 19 512 20 523 20 578 20 658 20 633 20 495 20 463 20 550 20 518 20 605 20 573 20 427 20 482 20 442 20 537 20 497 20 452 20 387 20 342 20 292 20 236 20 066 20 397 20 347 20 291 20 121 20 402 20 345 19 846 19 900 20 175 19 953 19 564 19 431 19 206 19 617 19 483 19 257 19 670 19 536 19 309 99 72 20 756 20 736 20 714 20 689 20 660 20 628 20 593 20 552 20 507 20 456 20 400 20 229 20 007 19 722 19 588 19 361 99 99 20 811 20 769 20 716 20 648 20 607 20 562 20 511 20 454 20 283 19 775 100 25 20 866 20 825 20 771 20 703 20 662 20 617 20 566 20 509 20 337 19 828 100 52 100 79 20 977 20 958 20 935 101 05 21 033 2
9. Fiber optic oxygen input for chamber 4 Connect an oxygen sensor e g oxygen dipping probe to an input LINE Indicates instruments ON OFF 3 4 2 Back side Power connector Power input for the OXY 4 instruments Connect the instrument to a grounded 230 VDC power supply using a standard pc type cable with a grounded wall plug Power button Turns instrument on and off DATA RS 232 interface male Connect the OXY 4 instrument to the PC via a RS 232 cable For further information about the OXY 4 fiber optic oxygen instrument see 8 4 OXY 4 Instruction manual page 50 Page 15 of 50 L oii o LOLIGOSYSTEMS COM 4 Introduction to intermittent flow respirometry Three different methods are commonly used for measuring oxygen consumption in water breathers e g fish e Closed respirometry e Flow through respirometry e Intermittent flow respirometry 4 1 Closed respirometry or constant volume respirometry Measurements are in a sealed chamber of known volume a closed respirometer The oxygen content of the water is measured initially t then the respirometer is closed and at the end of the experiment t the oxygen content is measured again Knowing the body weight of the animal the respirometer volume and the oxygen content of the water at time t and t the mass specific oxygen consumption rate can be calculated as follows VO O2 to O2 t V t BW VO oxygen consumption rate mg O kg hour
10. NaOH 90 C as well as a 3 H2O solution The oxygen sensitive material may be subject to photodecomposition resulting in a signal drift Photodecomposition takes place only during illumination of the sensor tip and depends on the intensity of the excitation light Table 5 4 Drift in air saturation at 100 air saturation when illuminating the oxygen sensor PS 3 for 1 12 and 24 hours in the continuous mode 1 sec mode Drift per hour Drift per 12 hours Drift per 24 hours PSt3 lt 0 4 air saturation Planar Oxygen Minisensor 10 5 2 Housings of Oxygen Sensitive Minisensors PreSens fiber optic oxygen sensors are based on 2 mm polymer optical fibers POF Depending on the respective applica tion PreSens offers a set of different standard designs cee Planar oxygen sensitive sensor foils Flow through cell design connected to 2 mm Dipping Probe DP with coated SP a 2 mm POF FTC sensor foil Of course it is possible to build customer specific designs Please feel free to contact our service team to find the best solution for your application Planar Oxygen Minisensor 11 5 2 1 Planar Oxygen Sensitive Foils SP PSt3 Planar oxygen sensors SP PSt3 immobilized onto different supports polyes ter glass are available for customer specific applications Sensors based on a polyester support can be easily cut into small pieces using a razor blade Round spots sensor spots of 3 mm in diameter can be punched
11. The sensor spots can be glued for example inside glass vials such as cell culture flasks bags and disposables The oxygen concentration can be measured non invasively and non destructively from outside through the wall Only prerequisite The wall has to be transparent and non fluorescent Please note Be sure to glue the sensor spots onto your vessel with the proper side The sensor support polyester foil or glass on which the sensor is spotted identifiable by its faint reflection is glued to the vessel while the sensor itself must look toward the sample The figure below shows how the highly enlarged sensor spot must be glued to the vessel Sample Optical isolation Sensor Sensor Support Vessel bottom Sensor spot SP glued into a vessel no optical isolation Optical isolated sensor spot SP glued into a vessel Ganapr Features foll inside e non invasive and non destructive measurement from outside through the wall of the flask e excellent mechanical stability and long term stability more than 100000 data points without drift e online monitoring e response time teo in the order of 30 s e measuring range 0 250 a s e limit of detection 0 15 a s Container wall Optical transmit signals ta analyzer Oxygen sensor immobilized onto a glass support e stands CIP Cleaning In Place conditions Optical receive e Ssterilizable autoclave 130 C 1 5 atm ethanol ethylene
12. e no longer operates as specified e has been stored under adverse conditions for a lengthy period of time e has been damaged in transport f you are in doubt the instrument should be sent back to the manufacturer PreSens for repair and maintenance The operator of this measuring instrument must ensure that the following laws and guidelines are observed when using dangerous substances e EEC directives for protective labor legislation e National protective labor legislation e Safety regulations for accident prevention e Safety data sheets of the chemical manufacturer The OXY 4 is not protected against water spray The OXY 4 is not water proof The OXY 4 must not be used under environmental conditions which cause water condensation in the housing The OXY 4 must not be opened We explicitly draw your attention to the fact that any damage of the manufactural seal will render all guarantee warranties invalid Any internal operations on the unit must be carried out by personnel explicitly authorized by PreSens and under antistatic conditions The OXY 4 may only be operated by qualified personnel This measuring instrument was developed for use in the laboratory Thus we must assume that as a result of their profes sional training and experience the operators will know the necessary safety precautions to take when handling chemicals Keep the OXY 4 power supply and optical sensors out of the reach of children As the manufacturer of th
13. lpoint x Lock LO EEE Input v 2 96 LO input ty Joo HI input V Jj 5 00 pO kPa 12 41 LO pO kPa 0 00 y HI pO kPa J 120 96 Temperature Mov Avg lpoint Lock LO j Lock HI Input V 2 72 LO input V Fo y HI input V Jj 5 00 Temp C 54 47 LO Temp eG 2 9 00 HI Temp C 0 1 Jl 100 00 13 42 20 Page 22 of 50 1 13 43 20 L oii o LOLIGOSYSTEMS COM 5 2 2 Sample frequency button Use this button to set a common sample frequency for all inputs The frequency can be set from one sample to ten samples per second Please note All inputs are always sampled at the same rate 5 2 3 Barometic pressure hPa Use this button to set the days actual barometic pressure in hPa This field is used only in fiber optic measurements and is disabled enabled together with the fiber optic button status OFF ON The default value is 1013 1 atm 760 mmHg 5 2 4 pO2 sat kPa Use this button to set the saturated partial pressure of oxygen in kPa see 8 1 pO2 kPa table on page 45 This field is needed to convert the oxygen values in kPa to air sat and oxygen sat via the Unit button see 5 2 11 2 Unit button on page 29 The default value is 20 859 which corresponds to 1013 kPa at 15 9c 5 2 5 Fiber optic button Use this button to select the type of oxygen input When the button displays Fiber optic off chamber oxygen input should be 0 5 VDC e g fro
14. 0 2 100 Zas gt 0 100 as gt 0 a s gt 100 as 2 i a s gt 100 as a s gt 100 as 8 2 Some Advice for Correct Measurement 8 2 4 Signal drifts due to oxygen gradients Please keep in mind that the sensor only measures the oxygen content near its surface In unstirred solutions an oxygen concentration gradient often occurs Please check if air bubbles are on the sensor tip whenever unexpected drifts gradients or unstable measurement values occur Critical conditions for bubble formations are for example purging with air or other gases and increasing temperature during measurement The formation of a bio film during long term measurements or the accumulation of other sample components like oil or solid substances may lead to an oxygen gradient 8 2 2 Signal drifts due to temperature gradients A further source of imprecise measurement is insufficient temperature compensation Please bear in mind that the OXY 4 only measures correctly if the sample temperature is constant during measurement and the temperature is the same as you typed in at the beginning of the measurement Please also refer to Chapter 12 5 Temperature Compensation of the Re sponse of Oxygen Sensors Measurement with Oxygen Sensor PSt3 45 8 2 3 Signal drift due to photodecomposition The oxygen sensitive material may be subject to photodecomposition resulting in a signal drift Photodecomposition takes place only during illumination of the
15. 01 05 11 33 10 057 3374 27 19 28 01 05 11 33 15 0 65 99 98 2747 28 01 05 11 33 20 074 100 14 27 15 gt HN test ch 44241 44245 44247 44239 44233 44247 44250 44254 44242 44234 TemprC Logged data of channel 1 displayed in Microsoft amp Excel amp 20 20 20 20 20 20 20 20 20 20 27 Calibration 28 7 Calibration The second window in the menu bar of the OXY 4 software is named Calibration It consists of two sub windows The sub window SINGLE CHANNEL Figure 7 1 is used to calibrate each channel individually The sub window ALL CHANNELS Figure 7 2 calibrates all channels with one command Use this option if you want to calibrate all channels in one calibra tion procedure Please note For calibrating the sensor it is necessary to stop the measurement Calibration during measurement is not possible To stop the measurement go to the window Measurement and click the All channel button if you perform a measurement with all channels or the respective active channel button During a measurement the calibration buttons are not active SINGLE CHANNEL ALL CHANNELS 2nd cal point pressure temperature of cal0 standard temperature of cal100 standard 09625 100 009625 ERE 56 00 E 27 00 Figure 7 1 Screenshot of the sub window SINGLE CHANNEL of the window Calibration SINGLE CHANNEL ALL CHANNELS Vine Ko 9 a 1100 00 ME CHANNEL pressure temperature o
16. 21 321 21 275 21 222 21 163 20 986 20 756 20 461 20 321 20 086 Page 45 of 50 S 9 QD E D e g g Salinity 07 00 Oxygen solubility in mg O2 liter kPa at different temperatures and salinities From Green amp Carrit 1967 J Mar Biol 25 140 147 1 kPa Constants 8 2 Oxygen solubility 7 424 4417 2 927 0 04238 0 1288 53 44 0 0444 0 00071 273 16 John Fleng Steffensen 2002 Temperature deg C 0 0 6976 genta qare 0 6685 esan Det D AME ML Pcr 2 3122 i acsi 8 9287 0 5883 0 5800 0 5638 p sies usen 0 5303 i o 6788 0 6694 o 6600 0 6509 0 6329 0 6241 0 6154 0 6068 0 5984 0 5900 0 5818 0 5737 0 5657 0 5501 0 5424 0 5349 0 5274 2 0 6608 pisi es 0 6252 geier 0 082 0 69281 0 5916 0 50241 0 57541 0 5678 0 5597 0 5520 0 5444 0 5369 0 5295 2 8223 0 8181 3 0 6436 0 6349 0 6263 0 6094 0 6011 0 5929 0 5849 0 5769 0 5691 0 5614 0 5537 0 5462 0 5388 0 5315 0 5243 0 5171 0 5101 0 5032 4 Hes 0 5783 0 5706 D 2623 p sss 0 5479 0 5405 0 5121 sona 0 4937 5 0 5953 0 5643 0 5558 0 5494 0 5421 0 5349 0 5278 0 5004 0 4872 0 4807 6 0 5963 0 5885 n 5808 0 5731 0 5656 0 85821 asenn 0 8436 p 365 0 3294 0 5229 0 5156 0 5089 0 5022 0 4956 0 4891 D acra Aden 0 4701 7 0 5818 0 5743 0 5668 0 5595 0 5522 0 54s0 0 5379 0 5310 0 5241 0
17. 3811 0 3757 0 3724 0 3681 0 3639 0 3597 0 3435 0 3356 0 3318 25 0 4014 0 3968 0 3923 0 3878 0 3833 EE EE REREEER 0 3497 0 3457 0 3417 0 3378 eaan 0 2263 26 0 3947 0 3902 0 3857 0 3813 0 3770 0 3684 0 3642 0 3601 0 3560 0 3519 0 3479 0 3439 0 3400 0 3361 0 3323 0 3248 0 3211 27 0 3882 0 3838 0 3794 0 3708 m REPRE 0 3423 0 3384 0 3346 0 3308 0 3270 ER 28 0 3819 0 3776 0 3733 0 3649 0 3567 0 3486 0 3447 0 3408 0 3369 0 3331 0 3293 0 3256 0 3219 0 3146 0 3110 29 0 3759 0 3717 0 3674 0 3633 0 3592 E 0 3471 piusgsaguxs 0 3317 0 3279 0 3242 0 3205 0 3169 B TECH 30 0 3701 0 3659 0 3618 0 3577 0 3537 0 3457 0 3418 0 3380 0 3341 0 3304 0 3266 0 3229 0 3193 0 3157 0 3121 0 3051 0 3017 Page 46 of 50 L oii o LOLIGOSYSTEMS COM 8 3 DAQ 4 Instruction manual The DAQ 4 instrument is used for data acquisition and relay controlling It is designed to run with the software AutoResp 4 but it can be used with other applications LIST OF PARTS DAQ 4 instrument AutoResp 4 Power Cord USB cable 4 fold extension cord for pumps qty 2 User manual SETUP To power up the instrument connect the power cord to the outlet 100 240 VAC 50 60Hz on the back side of the instrument Connect the USB cable to an USB port on your PC and to the outlet on the front side of the instrument named PC USING THE DAQ 4 FOR DATA ACQUISITION The DAQ 4 instrument acquires data on 6 channel
18. Both equations give values of 105 a which agree to better than 1 96 Appendix 63 Table 12 6 Values of the coefficients in equations 27 and 28 pense elie ENDE CMM cci siu Eqn 27 a 4 900 10 p 5 516 10 1 335 q 1 759 10 C 2 759 10 r 2 253 107 d 8 235 104 s 2 654 107 e 1 614 10 t 5 362 10 Eqn 28 A 7 424 ee I s on B 4 417 10 Q 5 344 10 C 2 927 R 4 442 10 D 4 238 10 S 7 145 107 Seawater has a typical salinity of 35 76 35 g 1000 g or a chloride content of about 19 o and therefore falls within the scope of both equations 12 5 Temperature Compensation of the Response of Oxygen Sen sors A typical oxygen response characteristic at different temperatures is shown in Figure 12 10 The phase angle is a function of the oxygen content tan f O2 and decreases with increasing the oxygen content 65 o dm m 0 596 0 5 0 0 0 0 5 0 5 id 1 5 15 0 5 55 T 1 596 T d 396 1 5 pem 6 r 3 45 6 2 6 6 12 o E 35 12 9 12 12 E 20 12 amp 25 ana 20 20 m 50 50 i52 3 50 sb 2 C 10 C 20 C 30 C 40 C 5 T T T T i T i 0 1000 2000 3000 4000 5000 time s Figure 12 10 Oxygen response characteristics at different temperatures Figure 12 11 displays the oxygen dependence of the phase angle at different temperatures and Figure 12 12 the respective Stern Volmer plots These two figures show that both the phase an
19. Channels Place the calibration standard 100 cal 100 containing wet cotton wool underneath the oxygen minisensor The vessel with the label cal 100 has to be closed with the screw top containing the hole Insert the plastic fiber carefully through the hole without touching the oxygen sensitive spot until it is about 3 cm deep inside the vessel Make sure that the plastic fiber with its sensor spot cannot touch the vessel and the cotton wool Calibration 31 C e LETS EARN SINGLE CHANNEL gt ALL CHANNELS Pe er A l A 100 00 por temperature of call standard temperature of cal100 standard Oxygen co enc air sat fF m j 99 90 d za 1013 Phase 27 191 Temp cal Amplitude 44265 Temp cali 00 A dialogue appears informing you that you will change the last calibration values Click the Continue button to store the new calibration data Repeat this procedure for all other channels by typing in the respective channel into the Channel Window to record the cal 100 value 5 To record the second calibration value oxygen free water cal 0 place the vessel with the label cal 0 underneath the oxygen minisensor Insert the plastic fiber about 2 cm deep into the cal 0 solution Make sure that the plastic fiber with its sensor spot cannot touch the vessel To increase the response time stir the cal 0 solution Wait about 3 minutes until the phase angle is constant the variation of the phase angle should be
20. DAGms to support the PCI 7041 5040E device is 8 1 if you need support for this device do not install this version of NI DAQmx Refer to the NI DAQ Readme for supported device models Traditional NI DAQ Legacy is an older driver with an outdated API that supports some additional older DAQ devices but not all of the latest DAQ devices Applications developed with earlier versions of NI DAQ will run under Traditional NI DAQ Legacy You can use both NI DAQms and Traditional NI DAQ Legacy in the same computer but must reset devices before changing from Traditional NI DAQ Legacy to NI DAQms Refer to the NI DAQ Readme for guidelines Support Removed from NI DAQ 7 0 and later NI DAQ 7 0 and later supports different devices operating systems application software and programming language versions than previous releases of NI DAQ Starting with version 7 0 NI DAQ device support excludes Vx DAQ Remote SCXI 1200 Series DAQ the DAQCard 500 700 family parallel port DAGPads and several other older devices Do not install NI DAQ 7 0 or later if your existing applications include unsupported components Refer to the NI DAQ Readme file on the distribution LOLIGOSYSTEMS COM Screen 3 3 Click Next Destination Directory Select the primary installation directory All software will be installed in the following location s To install software into a different location s click the Browse button and select another directory
21. Directory for amp utoRespl 4 C Program Files AutoResp 4 Directory for National Instruments products C Program FilesNational Instruments 5 Click Next Page 7 of 50 4 Select destination directory for AutoResp 4 and for the National Instruments driver LOLIGOSYSTEMS COM 6 If you accept the License Agreement please select I accept the License Agreement s Screen 4 License Agreement You must accept the license s displayed below to proceed RETURNS SHALL BE SUBJECT TO NI S THEN CURRENT RETURN POLICY 1 meanings NATIONAL INSTRUMENTS SOFTWARE LICENSE AGREEMENT INSTALLATION NOTICE THIS IS A CONTRACT BEFORE YOU DOWNLOAD THE SOFTWARE AND OR COMPLETE THE INSTALLATION PROCESS CAREFULLY READ THIS AGREEMENT BY DOWNLOADING THE SOFTWARE AND OR CLICKING THE APPLICABLE BUTTON TO COMPLETE THE INSTALLATION PROCESS YOU CONSENT TO THE TERMS OF THIS AGREEMENT AND YOU AGREE TO BE BOUND BY THIS AGREEMENT IF YOU DO NOT WISH TO BECOME A PARTY TO THIS AGREEMENT AND BE BOUND BY ALL OF ITS TERMS AND CONDITIONS CLICK THE APPROPRIATE BUTTON TO CANCEL THE INSTALLATION PROCESS DO NOT INSTALL OR USE THE SOFTWARE AND RETURN THE SOFTWARE WITHIN THIRTY 30 DAYS OF RECEIPT OF THE SOFTWARE INCLUDING ALL ACCOMPANYING WRITTEN MATERIALS ALONG WITH THEIR CONTAINERS TO THE PLACE YOU OBTAINED THEM ALL Definitions As used in this Agreement the following terms have the following 9 accept the License A
22. In the presence of oxygen the signal in our case the phase angle decreases The phase angle can be related to the oxygen content as shown in Figure 5 2 The theoretical aspects are explained more detailed in the appen dix 60 60 09790 0 O m 50 5 504 g 9 EJ 40 B 404 S o a o S oO i S 30 20 t 204 10 T 10 4 9998 0 2 4 6 8 10 12 14 16 18 20 0 20 40 60 80 100 120 140 160 180 200 220 240 time min air saturation Figure 5 1 Response of minisensor PSt3 toward Figure 5 2 Effect of the phase angle of minisensor changes in the oxygen concentration PSt3 on different oxygen contents Measuring range and limit of detection PreSens offers an oxygen sensor membrane called PSt3 which is tailored for oxygen measurements up to 250 air saturation The measuring ranges and the limit of detection of this sensor are given in Table 5 1 Table 5 1 Measuring range and limit of detection of the PSt3 oxygen sensor membrane Dissolved Oxygen Gaseous amp Dissolved Oxygen Measurement range 0 22 mg L ppm 0 250 96 air sat 0 50 oxygen sat 0 700 umol 0 380 Torr 0 500 hPa Limit of Detection LOD 0 15 air saturation 0 31 hPa 15 ppb dissolved oxygen 0 23 Torr Planar Oxygen Minisensor 7 Resolution and accuracy Since the oxygen calibration plot displays a non linear behavior the oxygen resolution is gi
23. and sixth row respectively The seventh row contains the temperature in C as given by the user Raw data can be used for user defined recalculations according to the formulas and tables listed in the appendix The data can be processed with any text editor The use of Microsoft Excel is possible by importing the file Figure 6 3 4 Software 23 01 05 11 32 Sw ver OQXY4v2 08 Header Ch 1 Description channel 1 100 a s gt 0 as IDENTIFICATION PHiboard number v1210406 PMnumber X 00000000 Serial number OWY 4 trace 1 03 003 MUX channel ON 01 PARAMETERS Signal LED current 030 Ref LED current 055 Ref LED amplitude 90242 Frequency 006 0 Sending interval 0001 Averaging 2 Internaltemp 20 0C SYSTEM SETTINGS APL function ON Temp compensation OFF Analogout OFF RS232echo OFF Oxygen unit a s CALIBRATION Sensortype Oxasphase 1 58 18 at 020 0 C amp 100500 100 00 a s phase 2 27 17 at 020 0 C amp 044800 Date ddmmuy 290105 Pressure mBar 1013 FIRMWARE Code 3 021 IAP 01 17 05 16 11 16 Xilins built 01 05 04 MMOD Y Pressure hPa 1013 Datefdd mm y Timefhh mm Logtimefmin Oxy air sat Phasel Amp 28 01 05 11 32 38 0 39 9 27 18 28 01 05 11 32 43 0 12 100 02 27 16 29 01 05 11 32 46 0 16 99 84 27 18 29 01 05 11 32 51 0 24 99 62 27 21 29 01 05 11 32 55 0 32 3374 27 19 29 01 05 11 33 00 04 33 65 272 28 01 05 11 33 05 048 99 93 27 17 28
24. at combining the best of both 1 c osed and 2 flow through respirometry Reference Steffensen J F 1989 Some errors in respirometry of aquatic breathers how to avoid and correct for them J Fish Physiol Biochem 6 49 59 The experimental animal is placed in a closed chamber respirometer immersed in an ambient tank A recirculation pump ensures proper mixing of the water inside the respirometer and adequate flow past the oxygen probe A second pump can change the water inside the respirometer with ambient water During measurements of oxygen consumption this flush pump is turned off and the systems operates like 1 closed respirometry Then the pc controlled flush pump turns on pumping ambient water into the respirometer and bringing the oxygen content back to pre measurement values In this way problems with accumulating metabolites and severe changes in oxygen level due to animal respiration are avoided As with open respirometry the duration of the experiment is in principle unlimited However the most important advantage is the great time resolution of this method Oxygen consumption rates of animals can be determined for every 10 minutes over periods of hours or days making the system extremely suited for uncovering short term variations minutes in metabolism In summary our systems for respirometry are developed for prolonged and automatic measurements of oxygen consumption rate in a controlled laboratory environment
25. button Oxygen unit Enter the sampling rates and the temperature for each channel Please note If you want to adjust different scanning rates for the activated channels ensure that the locked field is not activated 5 Start the measurement by clicking the required channel button 10 qw oz 8 1 3 Logging Measured data are logged during the measurement Add contents to the description fields in the File description section before starting logging This is not necessary but helpful as the description is later stored in the data file 1 Depending on the scan mode click on All Channels for logging all the selected channels in the measuring window or Single channel for single channel logging 2 Inthe appearing save as window specify filename without extension and its location The PC software automatically creates one to four different files The filename consists of the specified filename and the extensions ch1 txt ch2 txt to ch4 txt respectively All filenames are displayed in the section Created files Measwemert Caran Logos Aches I CREATED FILES SINGEL CHANNEL MAIN PATH 1 DXoxydv2 D2T Mest chl tet ALL ACTIVE 2 D oxy4v2_O2T Stest ch2 txt 12 3 4 CHANNELS _D hoxy4v2_O2T test 3 D oxp4y2_02T test ch3 txt f p P f 5 4 D Noxp4v2 2TStest ch4 t t JA JA JA A FILE Desenption ch 1 Description ch 2 Description ch 3 Desenption ch 4 DESCRIPTION channel 1 chi i channel 3 channel 4 100 as gt
26. chan nel or switch to the sub window All channels Enter the respective calibration values a phase at 0 96 air saturation temperature at 0 air saturation phase at 100 air saturation temperature at 100 air saturation pressure at which values a d were measured The atmospheric pressure of the calibration is needed to convert the oxygen unit 96 air saturation into par tial pressure units hPa Torr or concentrations units mg L umol L paap 0 2 100 00 5 Phase J 56 00 27 00 E To store the calibration values click the Calibrate button A dialog appears and informs you that you will program all channels in case the sub window All channels is chosen If sub window Single channel is chosen a dialogue appears informing you that you will change the last calibration values Click the Continue button to store the new calibration data LESEN PEE WARNING This will change last calibration values Cancel Repeat this procedure for all other channels by typing in the respective channel into the Channel Window to record user defined calibration values Measurement with Oxygen Sensor PSt3 43 8 Measurement with Oxygen Sensor PSt3 Calibration of the sensor is recommended before each measurement see chapter 7 If you don t want to recalibrate a sensor you can use the calibration values of your last measurement These values are stored in the hardware of OXY 4 Each calibration is
27. channels are enabled there will be no chamber oxygen values the first ten samples equivalent to the first twenty seconds of samples 5 2 13 2 Enable channel By clicking on this field the channel gets enabled and oxygen data will be received Please note It is only possible to enable a channel when a sensor is connected to the OXY 4 instrument Please note If a sensor is not used then disable the channel This will increase the lifetime of the sensor 5 2 13 3 pO kPa indicator This field displays the chamber oxygen pressure from the OXY 4 fiber optic instrument from PreSens relative the calibration 5 2 13 4 Lock LO button Press this button to calibrate the OXY 4 fiber optic oxygen instrument in a 0 air saturation O sample Please note It is important not to push the Lock LO button before the Phase 9 and Amplitude indicators has stabilized Please note A full calibration must consist of a 0 calibration and a 100 calibration before the oxygen readouts are reliable Please note The calibration will be written down on an EEPROM in the OXY 4 instrument If you want to use the OXY 4 instrument with the software from PreSens a new calibration may be done using this software 5 2 13 5 LO Temp C Use this field to specify the temperature during calibration of the OXY 4 fiber optic instrument with the 0 air saturation O solution Page 31 of 50 L oii o LOLIGOSYSTEMS COM 5 2 13 6 Lock HI button P
28. detection 0 15 96 a s Ordering information OXYFinger Oxygen Sensitive Coating PSt3 0 250 air saturation Order code for the OxyFinger OFG PSt3 17 Planar Oxygen Minisensor 18 5 2 7 Coaster for Shaking Flasks and Spinner Flasks CSF Application Online control of oxygen in shaking flasks and spinner flasks Control ES amp Data Coaster System set up for online DO measurement in shaking flasks Specifications The coaster for shaking flasks and Spinner flasks is a tool for online monitoring of dissolved oxygen concentration in shaking flasks The coaster for shaking flasks does not contain a sensor It redirects the light so that oxygen sensors in shaking flasks can be read out The coaster has a colored circle which makes it easy to position it right under the sensor The position of the optical read out is flexible and can be adjusted to different sizes of shaking flasks The standard cable length is 2 5 m The cable has an outer diameter of 2 6 mm Please note The coaster for shaking flasks and spinner flasks does not contain a sensing layer It is designed to read out sensor foils which are attached to the inner side of a shaking flask spinner flask or a similar vessel e g beaker Ordering Information Order code for the Coaster for Shaking Flasks CS Software 19 6 Software 6 1 Software Installation This software is compatible with Windows 98 2000 Millenium NT4
29. experiment This option stops the current experiment Please note To immediately turn on the flush pump use this option 5 4 8 2 Edit gt Oxygen While the experiment is running the user can use this option to alter the oxygen regulation This can be done by changing the setpoint the hysteresis or the mode hypoxic hyperoxic 5 4 8 3 Edit gt Temperature While the experiment is running the user can use this option to alter the temperature regulation This can be done by changing the setpoint the hysteresis or the mode cool heat 5 4 8 4 View By this option the user decides which graph is to be shown in the graph field see 5 4 3 Graph field on page 38 5 4 8 5 Help gt About Displays contact information about Loligo Systems Page 40 of 50 L oii o LOLIGOSYSTEMS COM 6 Troubleshooting 6 1 No connection to the DAQ 4 instrument Screen 26 Error 201003 occurred at DAQmx Create Channel DO Digital Output vi 1 Possible reason s Measurements Device cannot be accessed Possible causes Device is no longer present in the system Device is not powered Device is powered but was temporarily without power Device is damaged Ensure the device is properly connected and powered Turn the computer off and on again If you suspect that Fhe device ic damaned contact National Tnetriinments at The error shown on Screen 26 occurs when the NI USB 6215 DAQ card has no properly connection
30. measured temperature 5 4 5 3 Oxygen regulation This field shows if the oxygen regulator is working to maintain the oxygen level in the ambient tank as hypoxic or hyperoxic see 5 3 5 Oxygen control field page 35 5 4 5 4 Temperature regulation This field shows if the temperature regulator is working to maintain the temperature level in the ambient water see 5 3 6 Temperature control field page 36 5 4 6 Chamber measure field 1 4 5 4 6 1 Chamber O kPa This field shows the actual measured oxygen pressure for the chamber 5 4 6 2 MO field This field shows the calculated MO value for the chamber as described in background see 4 Introduction to intermittent flow respirometry page 16 The MO value is calculated as follows MO 8 Os _ 3600 0 solubility E02 i stope ZE ee e h kg l kPa 5 wet weight Where slope is calculated as follows E O B Od slope S time time Please note MO is only calculated in each measurement period e g 29 MO values after 30 seconds measure period at 1 Hz Page 39 of 50 L oii o 5 4 6 3 R field LOLIGOSYSTEMS COM This field shows the calculated R linear correlation coefficient value for the chamber when MO is calculated The R value can be used to validate the linear fit of the O curve 5 4 7 Graph Legend This field shows the different colors of the oxygen and temperature signal 5 4 8 Menus in Screen 25 5 4 8 1 File gt Stop
31. only valid for the corresponding sensor Especially after longer measurements more than 10000 meas ure points or 3 h continuous mode the sensor should be re calibrated Ensure that the temperature of the sample is known and is constant during measurement 8 1 Starting the measurement Note Saving of data is explained in chapter 6 3 3 page 22 and in chapter 8 1 3 page 44 Logging Measurement is started either for selected one to four channels or for all channels In both cases the scanning rate can be defined for each channel separately In both cases the channels are scanned sequentially Click the Measurement menu bar to adjust the measurement conditions 8 1 1 Scanning all channels Switch on OXY 4 and PC software following chapter 6 Go to window Measurement Choose oxygen measurement unit from button Oxygen unit Enter the sampling rates and the temperature for each channel By clicking locked the adjusted sampling rate of the first channel is also set for all other channels 5 Startthe measurement by clicking button A channels Aog qw o DATE TIME SAMPLING CHANNEL RATE TEMPERATURE 29 01 05 11 48 31 J 200 A 200 A 200 locked aj locked m ALL CHANNELS OXYGEN UNIT WARNINGS pc opum ae eee d Measurement with Oxygen Sensor PSt3 44 8 1 2 Scanning selected channels Switch on OXY 4 and PC software following chapter 6 Go to window Measurement Choose oxygen measurement unit from
32. parameter t f Oa 2 The OXY 4 uses the phase modulation technique to evaluate the luminescence decay time of the indicators If the luminophore is excited with light with sinusoidally modulated intensity its decay time causes a time delay in the emitted light signal In technical terms this delay is the phase angle between the exciting and emitted signal This phase angle is shifted as a function of the oxygen concentration The relation between decay time t and the phase angle is shown by the following equation tan 2n m frod a tan 2r fma T 3b t tan O f O2 3c t luminescence decay time o phase angle fmoa modulation frequency Appendix 54 In max I 5 0 5 10 15 20 25 30 time ps Figure 12 5 Schematic of the single expo nential decay to gt t1 5 0 5 10 15 20 25 30 time us Figure 12 6 The luminophore is excited with sinusoidally modulated light Emission is delayed in phase expressed by the phase angle F relative to the excitation signal caused by the decay time of the ex cited state The measurement of the luminescence decay time an intrinsically referenced parameter has the following ad vantages compared to the conventional intensity measurement e The decay time does not depend on fluctuations in the intensity of the light source and the sensitivity of the detector e The decay time is not influenced by signal loss caused by fiber
33. sensitive dye at its inner wall The volume for liquid inside the FTC cell is about 100 10 ul The standard T shape flow cell can be easily connected via Luer Lock adapt ers to external tubings Liquids like water blood etc can be pumped through the cell Schematic drawing of flow through cell oxygen sensors Luer Lock adapter planar oxygen sensor Features female POF polymer optical fiber L 2 5 m Luer Lock adapter female SMA connector e very robust sensor with an excellent long term stability more than 100000 data points without drift e online monitoring e sterilizable autoclave 130 C 1 5 atm ethanol ethylene oxide e response time too in the order of 1 minute e measuring range 0 250 96 a s e limit of detection 0 15 a s e stands CIP conditions cleaning in place 5 NaOH 90 C Ordering Information Flow Through Cell with inte grated planar sensor Oxygen Sensitive Coating PSt3 0 250 air saturation Order code for the Oxygen Dipping Probe DP PSt3 Planar Oxygen Minisensor 5 2 3 Oxygen Dipping Probe DP PSt3 This oxygen sensor consists of a polymer optical fiber with a polished distal tip which is coated with a planar oxygen sensitive foil The end of the polymer optical fiber is covered with a high grade steel tube to protect both the sensor mate rial and the POF Usually the fiber is coated with an optical isolated sensor material in
34. sensor tip and depends on the intensity of the excitation light Therefore the excitation light was minimized Continuous illumination of a DP PSt3 oxygen sensor over a period of 24 hours may lead to a phase drift of up to 0 4 air saturation measured at 100 air saturation at 20 C However this effect of photodecomposition can even be minimized by changing the measuring mode to the second or minute interval mode In these modes the software switches off the excitation light after recording the data point and switches it on after the interval you have chosen Please use the interval method whenever it is possible to increase the shelf life of the minisensor Drift in air saturation at 100 air saturation when illuminating the oxygen sensor PS 3 for 1 12 and 24 hours in the con tinuous mode Drift per hour Drift per 12 hours Drift per 24 hours PSt3 lt 0 4 air saturation 0 air saturation phase angle 100 96 air saturation 0 5 10 15 20 25 time h Photodecomposition test of PSt3 continuously illuminating the sensor membrane for 25 hours Measurement with Oxygen Sensor PSt3 46 8 2 4 Signal drift due to too much ambient light A source of error is the detector overload due to too much ambient or sensor light A detector overload can be recognized with appearing warning prospective the warnings are activated Please notify that your measurement is not reliable if
35. signals to analyzer oxide H202 Oxygen sensor immobilized onto a polyester support e stands CIP Cleaning In Place conditions e Sterilizable ethanol ethylene oxide H202 e not autoclavable e flexible Planar C Minisensor A polymer optical fiber is used as a light guide between the OXY 4 oxygen meter and a sensor foil SP PSt3 which was glued inside a glass vial to read out the analyte concentration non invasively and non destructively from outside through the transparent wall of the flask Ordering information NUES Oxygen Optical Isolation Spot Diameter in mm Sensor Spot Y with optical isolation E gt N without optical isolation 10 Oxygen Sensitive Coating Support PSt3 0 250 air saturation PE polyester support G glass support Example oe dl E ed HH With this code you will order a planar oxygen sensor spots type PS 3 0 250 air saturation with optical isolation YOP immobilized onto a glass support G SUP The spot diameter is 5 mm Planar Oxygen Minisensor 13 5 2 2 Flow Through Cell with Integrated Planar Oxygen Sensor FTC PSt3 The flow through oxygen minisensor FTC PSt3 is a miniaturized fiber optic chemical sensor integrated in a T shape flow through cell The flow through cell is connected to the PreSens oxygen meter OXY 4 by a polymer optical fiber with 2 mm diameter as a light guide A glass tube with 2mm inner diameter 4 mm outer diameter is coated with oxygen
36. the warning light overload is shining red A detector overload can affect both the amplitude and the phase angle Please decrease the LED intensity to reduce the amounts of photons reaching the photodetector 8 2 5 Performance proof If you want to prove the performance during the past measurement please check the calibration values by inserting the sensor tip in the cal 0 and cal 100 calibration standards when you have finished your measurement If the device shows 0 air saturation immersing the sensor tip into the cal 0 solution and the value of the second calibration point measuring the cal 100 standard the sensor worked perfectly during the whole measurement 8 2 6 Correction for air pressure variations The atmospheric pressure of the calibration is needed to convert the oxygen unit air saturation into partial pressure units hPa Torr or concentration units mg L pmol L The partial pressure and the oxygen concentration units are calculated from air saturation by the software Consequently changes in the actual atmospheric pressure have no effect on the partial pressure units hPa Torr and concentration units mg L umol L but the oxygen units 96 air saturation and 96 oxy gen saturation have to be corrected for air pressure changes General Instructions 47 9 General Instructions 9 1 Warm Up Time The warm up time of the electronic and opto electronic components of the OXY 4 is 5 min Afterwards s
37. there will be no ambient oxygen values the first 30 samples equivalent to the first three seconds of samples 5 2 9 2 Input V indicator This field displays the actual voltage input 5 2 9 3 pO kPa indicator This field displays the actual calculated oxygen partial pressure from the voltage input 5 2 9 4 Lock LO button This button can be used to copy the actual value from the Input V indicator to the LO input V field instead of manually reading and retyping the value 5 2 9 5 LO input V Use this field to enter the low input voltage from the Input V indicator during the ambient oxygen calibration Please note The field is limited to values between OV to 5V Page 26 of 50 L oii o 5 2 9 6 LO pO kPa LOLIGOSYSTEMS COM Use this field to enter the oxygen pressure during the low level oxygen calibration of the ambient oxygen input Please note The field is limited to values between 0 00 kPa to 60 00 kPa 5 2 9 7 Lock HI button This button can be used to copy the actual value from the Input V indicator to the HI input V field instead of manually reading and retyping the value 5 2 9 8 HI input V Use this field to enter the high input voltage from the Input V indicator during the ambient oxygen calibration The lower Lock button can be used to copy the actual value from the Input V indicator to the HI input V field instead of manually reading and retyping the value Please note The field i
38. to the PC This error can occur in 2 ways 6 1 1 Name is not Devi To change the device name open Measurement amp Automation by following the folder hierarchy shown on Screen 15 page 13 The following example shows on Screen 27 a simulated device named as Devi The NI USB 6215 DAQ card is registered as Dev2 Page 41 of 50 LOLIGOSYSTEMS COM X A iSi Properties amp il Data Neighborhood amp filii Devices and Interfaces amp illl NI DAQmx Devices W NIUSB 6215 Dev1 NIUSB 6215 Dev2 PXI PXI System Unidentified X Delete o Self Test HA Test Panels Ja Reset Device Name Value Diserial Number 0x0 NI DAQGmx Device Basics aA scales a amp Software E Q Remote Systems What do you want to do Bun the NI DAQmx TestPanels Remove the device View or change device properties m Y sareen 18 bmp Paint Cyr use e215 Devi s X us DEVO SS 53am Right click with mouse on the Dev1 and choose rename or press F2 Change the name to something different than Dev1 In this example the name is changed to devisimu see Screen 28 Screen 28 File Edit View Tools Help TA i Properties X Delete Zh Self Test Wy Test Panels J Reset Device ai Create Task a Configure TEDS y Hide Help
39. touch the vessel and the cotton wool Measurement Calibration Logging All channels d and eal nni Se air 2nd cal point 100 00 2 EA 1013 7 temperature of call standard s 200 9X temperature of cal100 standard E 200 Wait about 3 minutes until the phase angles are constant and press the CAL 100 button to store the 100 air sat calibration values at the adjusted temperature A message window opens and informs you that you will program all channels with the actual calibration values Click the OK button to store the new calibration data To record the second calibration values oxygen free water cal 0 place the vessel with the label cal 0 underneath the oxygen minisensors Insert the plastic fibers about 2 cm deep into the cal 0 solution Make sure that the plastic fibers with their sensor spots cannot touch the vessel To increase the response time stir the cal 0 solution Wait about 3 minutes until the phase angles are constant the variation of the phase angles should be smaller than 0 05 and click the CAL 0 button to store the 0 calibra tion values at the adjusted temperature A message window opens and informs you that you will program all channels with the actual calibration values Click the OK button to store the new calibration data Rinse the sensor membranes gently with water after calibration to remove sulfite remaining on the surface Calibrate Single
40. 0 XP 1 Insert the supplied disc CD into the respective drive Start the file setup exe in folder installer and install the pro gram to the desired folder 2 Thereafter Labview runtime engine is installed Please click Next in the following three windows without chang ing the chosen options 3 Finally click Finish in the next window 1 amp National Instruments LabVIEW Run Time Engine 6 0 Setup NATIONAL INSTI Feature Description LabVIEW Run Time Engine Modify Feature Description gr Change which application features are installed Displays the LabVIEW Run Time Engine ee Select Features dialog which lets you configure individual features o This feature will remain on your local hard This feature will remain on your local hard drive drive O Repair Reinstall missing or corrupt files registry keys and shortcuts AT Preferences stored in the registry may be reset to default values This feature requires OKB on your hard drive This feature requires 0KB on your hard drive ius Uninstall National lr LabVIEW Run Time Et 6 0 fror CAProgrammelNational Instruments CAProgrammeNational Instruments y piskcos __ Reset Beck New gt Cancel Disk cost Figure 6 1 Screenshots of the windows appearing during the installation process of Labview Run time Engine lt Back Finish Figure 6 2 Screenshot of the last window of the installation o
41. 1 12 1 Basics in Optical Sensing of Oxygen eeeeeseseeeeeeeeeeeeee nennen entente nnn nnn nnn n nnne nnn 51 12 1 1 Dynamic Quenching of Luminescence sssssssssseseeeeeeee nennen nennen nnne nnne nnn 51 12 1 2 Major Components of Fiber Optic Minisensors eesseseeeeenneeeeneenn enne 52 12 1 8 Advantages of Optical Oxygen Sensitive Minisensors esseneenneneen 53 12 1 4 Luminescence Decay Time eseeeseessesieseeeee eene enne nnnm nannten en nnnn entere enne enters 53 12 1 5 Literature mer i er aei ete decente aee te ad ad cedens 54 12 2 Determination of the Oxygen Concentration Using a Modified Stern Volmer Equation 55 12 3 Oxygen Conversion Formulas eeeeeeeeeee esee nennen nnne ninth n nnnn nter ane nn stis neta nenas nnns 56 12 4 Temperature Dependent Constants Affecting the Oxygen Content esses 58 12 4 1 Water Vapor Pressure eene eie dioe tec ette pe Dee eee ere trece EOD tne nh 58 12 4 Bunsen Absorption Coefficient enne enne nnne nnne nnne ntes ennnr en 59 12 4 8 Dependence on the Salt Concentration sss 62 12 5 Temperature Compensation of the Response of Oxygen Sensors eene 63 mE ENT 1 Preface Congratulations You have chosen a new innovative technology for measuring oxygen The OXY 4 is a compact easy to transport
42. 1 013 20 991 101 32 20 922 21 088 21 069 20 880 20 910 20 965 21 046 21 021 20 826 20 881 20 849 20 937 20 904 20 992 20 959 20 758 20 813 20 868 20 923 20 882 20 836 20 717 20 671 20 620 20 563 20 772 20 726 20 675 20 618 20 445 20 827 20 781 20 730 20 672 20 499 20 784 20 727 20 391 20 221 20 274 20 553 20 328 19 881 19 933 19 797 19 568 19 986 19 850 19 620 20 039 19 902 19 672 101 59 21 144 21 124 21 102 21 076 21 047 21 015 20 978 20 937 20 891 20 839 20 782 20 607 20 381 20 092 19 955 19 723 101 85 21 199 21 157 21 102 21 070 21 033 20 992 20 946 20 894 20 836 20 661 20 435 20 144 102 12 21 255 21 212 21 187 21 158 21 125 21 088 21 047 21 000 20 948 20 891 20 715 20 488 20 197 102 39 102 65 21 366 21 346 21 323 102 92 21 421 21 401 21 378 21 310 21 268 21 242 21 297 21 353 21 213 21 180 21 268 21 235 21 323 21 290 21 143 21 102 21 055 21 198 21 253 21 003 20 945 21 157 21 110 21 058 21 000 20 824 21 212 21 165 21 113 21 054 20 878 20 770 20 542 20 595 20 649 20 250 20 303 20 355 20 164 20 216 19 930 19 982 103 19 21 477 21 457 21 434 21 408 21 379 21 346 21 308 21 267 21 220 21 167 21 109 20 932 20 702 20 408 20 269 20 034 103 45 21 532 21 512 21 489 21 463 21 434 21 401 21 363
43. 10 5 LO input V Use this field to enter the low input voltage from the Input V indicator during the low temperature calibration Please note The field is limited to values between OV to 5V 5 2 10 6 LO Temp C Use this field to enter the temperature during the low temperature calibration Please note The field is limited to values between 0 C to 100 C 5 2 10 7 Lock HI button This button can be used to copy the actual value from the Input V indicator to the HI input V field instead of manually reading and retyping the value 5 2 10 8 HI input V Use this field to enter the high input voltage from the Input V indicator during the high temperature calibration Please note The field is limited to values between OV to 5V 5 2 10 9 HI Temp C Use this field to enter the temperature during the high temperature calibration Please note The field is limited to values between 0 C to 100 C Page 28 of 50 L oii o LOLIGOSYSTEMS COM 5 2 11 Chamber Oxygen graph field This graph shows the oxygen pressure vs time measured for the 4 chambers going back 1 min By right clicking the graph it is possible to show hide the output data from a chamber It is also possible to show hide the ambient oxygen This is only possible if the Play button is ON When OFF and right clicking it is possible to copy data and export a simplified image of the graph Right to the graph there are six fields where current data
44. 1024x768 pixels is required e AutoResp 4 communicates with the DAQ 4 instrument via the accompanying USB cable which must be connected e AutoResp 4 can handle two different types of oxygen inputs Either 0 5 VDC analog input OXY CH1 4 or serial inputs when using fiber optic oxygen instruments manufactured by PreSens e If chamber oxygen is measured using the OXY 4 fiber optic oxygen instrument one RS232 male DB9 connector must be available on the PC If not available use an USB 232 adapter e g USB Serial Adapter manufactured by VSCOM e If using the OXY 4 instrument turn this on before starting AutoResp 4 Input labels see Table 1 Table 1 DAQ 4 instrument label AutoResp 4 signal name OXY CH1 Chamber oxygen 1 OXY CH2 Chamber oxygen 2 OXY CH3 Chamber oxygen 3 OXY CH4 Chamber oxygen 4 OXY AM Ambient oxygen TEMP Temperature Output labels see Table 2 Table 2 DAQ 4 instrument label AutoResp 4 indicator name FLUSH Flush RECIRC Recirc D2 N2 Ambient O kPa TEMP Temperature 9C Page 3 of 50 L oii o LOLIGOSYSTEMS COM 3 2 DAQ 4 instrument 3 2 1 Front side OXY CH1 Analog oxygen input for chamber 1 The input range is 0 5 VDC OXY CH2 Analog oxygen input for chamber 2 The input range is 0 5 VDC OXY CH3 Analog oxygen input for chamber 3 The input range is 0 5 VDC OXY CH4 Analog oxygen input for chamber 4 The input range is 0 5 VDC Connect analog oxygen inputs e g from an OXY REG via pi
45. 140 160 180 time s Figure 5 3 Response characteristic of an optical isolated oxygen sensor PSt3 in a stirred and a non stirred sample solution and in the gas phase Table 5 3 Response times too of the oxygen sensors membrane PSt3 dissolved oxygen gaseous oxygen stirred not stirred oxygen sensor PSit3 too without optical isolation 20s 40s 6s too with optical isolation 30s 60s 10s Planar Oxygen Minisensor 9 Optical isolation Optical isolated sensor tips are required if your sample shows intrinsic fluorescence between 540 700 nm Consequently an optical isolation is recommended measuring in whole blood urine or chlorophyll containing samples Using optical iso lated sensors excludes the impact of colored samples and ambient light on measurements Furthermore the optical isola tion layer is applied to exclude strong ambient light to improve chemical resistance especially against oily samples as well as to reduce bio fouling on the sensor membrane Optical isolated sensor tips of oxygen sensors enable measurement in photosynthetically active samples since stimulation of photosynthesis due to emission of blue green light from the fiber tip is avoided PreSens offers additional optical isolation for all types of oxygen sensors Sensor Stability The oxygen sensitive membrane stands gamma sterilization sterilization by ethylene oxide steam autoclavation 140 C 1 5 atm CIP conditions cleaning in place 5
46. 2 2 LR RUE a aa a edi ER RR abe 43 8 1 2 Scanning selected channels ute edd tue e e ate ee ee dede 44 8 1 3 Logglfig epp Een Dude 44 8 2 Some Advice for Correct Measurement eeeeeeeeeeeeseeeeseeee eee en nennen nennen nnne nnne n nnne n nnn 44 8 2 1 Signal drifts due to oxygen gradients ssssssssseeeeeeeeenenenee nennen 44 8 2 2 Signal drifts due to temperature gradients sssssssseseeeneeneeeneneee nennen 44 8 2 8 Signal drift due to photodecomposition sssessseeeeeeeeneeeneneee nnns 45 8 2 4 Signal drift due to too much ambient light 46 8 2 5 Performance proof s une eaa eoe nn e Ee enn E 46 8 2 6 Correction for air pressure variations sesssssssseeseeeneeeeeennneen nene nennen nnns 46 9 General Instr ctlorns einn dcin Een nece ie eio ete erect aee 47 9 1 LEnit aur 47 9 2 l CIncnrucc araea aaRS 47 9 3 Ila 47 10 Technical Data sick HNIC cH M 48 Table of contents 10 1 cre MIicm 48 10 2 Technical Notes E 49 10 3 Operation rs 49 TA Concluding Rematrks iure Leere Ie aceetesesntusvaceesst sendtesunesnpetetaccncauesess 50 12 APP ix qu in 5
47. 24 23 21 20 19 18 33 18 17 15 14 13 12 11 09 08 07 06 34 06 05 04 02 01 00 99 98 97 96 94 35 6 94 93 92 91 90 89 88 87 85 84 83 36 83 82 81 80 79 78 77 75 74 73 72 37 72 71 70 69 68 67 66 65 64 63 61 38 61 60 59 58 57 56 55 54 53 52 51 39 51 50 49 48 47 46 45 44 43 42 41 40 41 40 39 38 37 36 35 34 33 32 31 Example cs 20 0 C 9 08 mg L Appendix 62 12 4 3 Dependence on the Salt Concentration Table 12 4 gives values of the concentration of dissolved oxygen at several temperatures in solutions with vari ous chloride concentrations Increasing the salt concentration leads to a decrease in oxygen solubility This be havior is characteristic for the solubility of many nonelectrolytes it is the phenomenon known as the salting out effect Instead of chlorinity CI the amount of chloride in parts per thousand which was used as a measure of the amount of salt in water the term salinity is often used If salinity is preferred as a measure of salt concentration then the conversion from g L can be readily made using equation 26 S 1 805 CI 0 03 26 where S is the salinity in o or g 1000g Table 12 4 Solubility of oxygen in water as a function of temperature and salt concentration Total pressure 760 torr T C Oxygen solubility mg L CI g 1000g 0 4 8 12 16 20 0 14 5 13 9 13 3 12 6 10 11 3 10 8 10 4 9 9 9 5 9 0 8 5 8 1 78 7 4 7 0 6 7 64 6 1 The effect of increasing the salt concentr
48. 4 2 Mua vemm n QXRAR LALAO Be orn Please note By changing the NI USB 6215 device to devi another applications using devices from National Instruments might not work properly anymore 6 1 2 Another USB device was improperly removed If an USB device like a USB memory stick is not properly removed while AutoResp 4 is running there can be a connection problem with the DAQ 4 instrument To solve this problem close AutoResp 4 Then disconnect the USB cable between the PC and the DAQ 4 instrument Wait 30 seconds Then connect the cable and wait When the green diode on the front side of the DAQ 4 instrument is flashing the connection is re established Choose Continue experiment to avoid loosing data 6 2 Relays are not working Make sure the DAQ 4 instrument is connected with a power cable and the Power button is ON Page 43 of 50 7 Index TG ect na ues A MI LE 31 32 Ambient O eese 39 Amplitude esses 32 Anal Csi T m 4 Chamber os usi nta t nie 39 CheckboX MEE HC ES 35336 Closed respirometry 16 COOL Tea scii Eb mb Rd 36 DAQ e NN RNC LR E 3 Rs C 14 41 Device nalTiG asi mcd pii RR biting 41 aon m 10 20 DIV EF vesciasissnatioaveesianevaneaseseseionaniens 7 10 Edit 2 Oxygen sss 40 Edit gt Temperature 40 Enable channel
49. 5172 0 5105 0 5039 0 4974 0 4909 0 4845 0 4782 0 4720 0 4659 0 4598 a 0 5680 0 5607 0 5535 0 5463 0 5393 0 5324 0 5255 0 5138 0 5121 0 5055 0 4990 0 4926 0 4558 0 4500 9 0 5547 0 5476 0 5406 0 5338 0 5203 0 5136 0 5071 0 5006 0 4943 0 4880 0 4818 0 4461 0 4405 10 0 5419 0 5351 0 5283 0 5151 0 5086 0 5022 0 4959 0 4896 0 4834 0 4774 0 4713 0 4654 0 4595 0 4537 0 4480 0 4368 0 4313 11 0 5165 oiu 0 485 0 4790 BA 0 4613 ime 0 4224 12 0 5052 0 4806 0 4747 0 4688 0 4630 0 4573 0 4516 0 4191 0 4139 13 0 5067 0 5005 0 4943 0 4823 D 0S 0 4647 D 159010 4524 0 4478 0 4423 0 4369 0 4315 0 4262 0 4210 ines 14 0 4959 0 4898 0 4839 0 4721 0 4607 0 4551 0 4496 0 4441 0 4387 0 4333 0 4281 0 4229 0 4177 0 4126 0 4026 0 3977 16 0 4641 0 4423 0 4370 0 4317 0 4266 0 4214 0 4164 0 3873 0 3827 17 0 4659 0 4603 0 4549 0 4494 0 4441 oue nar Dp nena 0 4084 0 4035 0 3987 0 3940 0 3893 Hann ams 18 0 4567 0 4513 0 4459 0 4407 0 4354 0 4252 0 4202 0 4152 0 4103 0 4054 0 4006 0 3955 0 3912 0 3866 0 3820 0 3730 0 3686 20 0 4291 0 4094 0 4046 0 3999 0 3952 0 3906 0 3860 0 3597 0 3555 21 0 4311 0 4261 0 4212 0 4163 0 4114 ERE 23926 0880 0 3535 0 3791 0 3747 0 3703 0 3660 0 3617 E 0 2403 22 0 4233 0 4184 0 4135 0 4087 0 4040 0 3947 0 3901 0 3856 0 3812 0 3767 0 3724 0 3681 0 3638 0 3596 0 3554 0 3473 0 3432 24 0 3991 0
50. 7 OXY citi ests ces tates 30 Oxygen partial pressure 16 22 Oxygen regulation 39 aote qe c 22 Phase totum MAIN ME 32 28 M E 3 pO lower control 20 27 pO upper control 25 27 Bu cub ELE E eat 40 Raw Ca ESI iocos sso bu pl b pF ins 24 33 RECIEC ceo pa Du beu Ms 38 Relays are not working 43 RS2 P M cU 3 Sample frequency 23 Setpoint eio ede EDS Ko 35 36 Start experiment ss 36 Steady SEaEG ie urs Iro piii phis 17 SESS MNT 16 Temp lower control 28 Temp upper control 28 Temperat Esas vatibus bae 39 Temperature regulation 39 Time resolution 17 18 Troubleshooting s 41 Dao 40 Voltage input 25 26 27 VOIUME coit timi SOME neo hii 35 Wait period uussensskice enin ercndisab nto 34 Wet welght eseecsitre trot rines 35 Page 44 of 50 S 9 QD E D e g z 8 Appendix pO kPa table 8 1 File P02 kPa xls Partial pressure of oxygen p02 at different barometric pressures and temperatures Pbp Pva 2094 Temperature deg C 97 32 20 256 20 237 20 215 97 59 20 312 20 292 20 191 20 271 20 246 20 163
51. A DE AENEA ENSE 20 5 1 ccm M H M 20 5 2 Calibration and selection of oxygen input cece eee cece eee eee eee teens tees teen teens 22 5 3 erubuit LEES 33 5 4 R nning experlTirit issue tusasue ntur Kr ER rexi idu RR RERMRERNRRINEMRRRANEEATNEKUNTARRRUNERENENRK EIUS 37 6 Troubleshootikig xerit tex enun ehe FR Rub X rna nERIR RERERYSERE barges AU EEERMRA REDE RE TAKE AEN RE 41 6 1 No connection to the DAQ 4 instrument csssssssseeseeeeen nne 41 6 2 Relays are not WOFKII s iisece ntur n tarn ernan Neen AER RERYEER RERFREREREETEKRERERAS EAE EREREKTEEEEKE 43 II CRT E E 44 APDENGIXK 2 cones cet EERR 45 8 1 pOs KPa table idv udo E A ERN EN UU EM EEE ER M NE VE T 45 8 2 Oxygen solubillby 1 courant n Rh RERERER TER ENR A FR AORXR stark enue CREE RENE EE 46 8 3 DAQ 4 Instruction MANUAL nnne menn ren nnn 47 8 4 QXY 4 Instr ction manual cus enn un rine na neces x IR EX Ex n ERE n RE REUFRUA AERE NEERAEK E ERTA EIE A 50 Page 2 of 50 L oii o LOLIGOSYSTEMS COM 3 Installation procedure for DAQ PAC F4S 3 1 General e AutoResp 4 is a piece of PC software designed to perform automated intermittent flow respirometry together with the DAQ 4 instrument e AutoResp 4 requires Windows XP running on a PC with an Intel Pentium IV processor of minimum 2 66 GHz or equivalent The PC must have at least 256 MB memory and at least 1GB of free hard disk space A minimum resolution of
52. CE Communication Protocol Instrument output serial interface RS 232 38400 Baud Databits 8 Stoppbits 1 Parity none Handshake none on RJ11 4 4 plug RJ11 4 4 48 Technical Data 49 Interface cable to PC RJ11 4 4 to DSub9 D Sub9 female 2 RXD 3 TXD 5 GND 1 4 6 7 8 9 n c ENVIRONMENTAL CONDITIONS Operating temperature Storage temperature Relative humidity up to 95 OPERATION CONTROL LED at the front panel DIMENSIONS length 300 mm width 250 mm height 140 mm weight 3600 g 10 2 Technical Notes The unit uses a special interface cable Another cable can cause the unit s malfunction Optical Output SMA The SMA connector is a high precision optical component Please keep it clean and dry Always use the rubber cap to close the output when not in use 10 3 Operation Notes To achieve the highest accuracy the OXY 4 should be warmed up for 5 min before starting the measurement Please see the details of the measurement process described in the OXY 4 manual Concluding Remarks 50 11 Concluding Remarks Dear customer With this manual we hope to provide you with an introduction to work with the OXY 4 fiber optic oxygen meter This manual does not claim to be complete We are endeavored to improve and supplement this version We are looking forward to your critical review and to any suggestions you may have You can find the newes
53. Calibrate button A dialog appears and informs you that you will program all channels in case the sub window all channels is chosen If sub window single channel is chosen a dialogue ap pears informing you that you will change the last calibration values Click the Continue button to store the new calibration data DENEN WARNING l This will change last calibration values Cancel Repeat this procedure for all other channels by typing in the respective channel into the Channel Window to record user defined calibration values Calibration 33 7 2 Calibration of Flow Through Cell FTC PSt3 There are two ways to calibrate the optical sensors One is the use of two calibration standards the other one is to type in known constants The first way needs two calibration standards Their preparation is described in chapter 7 2 1 The second possibility is described in chapter 7 2 2 7 2 1 Calibration with two calibration standards For the calibration with current values you need calibration standards and your mounted sensor 7 2 1 1 Preparation of the Calibration Standards Calibration of oxygen minisensors is performed using a conventional two point calibration in oxygen free water cal 0 and water vapor saturated air or air saturated water cal 100 In the sub window Calibration cal 100 is denoted as 2 cal point Preparation of calibration standard cal 0 oxygen free water 1 Add 1 g sodium sulfite Na2SO to the vessel an
54. L oii o LOLIGOSYSTEMS COM USER MANUAL DAQ PAC F4S 1 List of parts e DAQ 4 instrument O OOoo0Oo O0 AutoResp 4 Power Cord USB cable 4 fold power strips for pumps qty 2 User manual e OXY 4 fiber optic oxygen instrument OOo 0 Power Cord RS232 cable User manual PC operating software for Windows 98 00 ME NT XP e Input connectors qty 2 e Device connectors qty 2 Page 1 of 50 L oii o LOLIGOSYSTEMS COM 2 Contents List Of Parts comaoine oina Dra oig dis heben rk nive RR E OER RERO RV E nets 1 2 Sp 2 Installation procedure for DAQ PAC FAS ssssssssssssseeseseenn ensem emen nens 3 3 1 General minnein nrn eda pbRRI VR D RI PGERIDPL DR ascot REPAS DUK OPEP MPERDRDEDITXPIT E KNEE ETAT 3 3 2 DAQ 4 instr rment e cectexcesvecerv ees eerie epi bins eripe ras eddies bei O EEEO Cri Der PE Da Y Cad d 4 3 3 AutoResp 4 software for Windows sssssssssssssessen emen meses mese nene 6 3 4 OXY 4 fiber optic oxygen instruments sss mener 15 4 Introduction to intermittent flow respirometry sss enne 16 4 1 Closed respirometry or constant volume respirometry e cece eee e eee ee teens ee eees 16 4 2 Flow through respirometry or open respirometry cceeeee eee ee eect eee e eee eaten ee 17 4 3 Intermittent flow respirometry or open closed respirometry sssusse 18 5 USINGJAUTORES DP 4 arrar iaa vara end xe ce easiest EEEE ETA A
55. NELS m OXYGEN UNIT WARNINGS t Oxygen Amplitude Clear graph auto scale p Channel 1 settings 4 mm A 2 22 0 7 p 6 3 4 Measurement With this software it is possible to start the measurement scanning all channels by clicking the All Channel button or se lected channels by clicking the required single channel button Before starting the measurement you have to choose the oxygen unit air saturation hPa Torr mg L or umol L the sampling rate fastest sampling max 15 s and enter the measurement temperature The button Warnings has the possible captions WARNINGS ON This caption indicates that the software will warn if no sensor is connected or if the sensor intensity is low If sensor intensity is low the ntensity check should be done by the user If the sensor intensity process fails the sensor must be replaced WARNINGS OFF This caption indicates that the software will show no warnings This is only recommended for advanced users On the right side you can find the version number of the software If you have questions regarding the OXY 4 software please contact our service team and have the software version number ready Furthermore the actual date time are presented on the right side 6 3 2 Calibration The detailed calibration process of oxygen sensors you can find
56. O mg L Uu OQ CO oO 20 25 0 C Figure 12 9 Dependence of the oxygen solubility in air saturated fresh water on temperature Table 12 3 Oxygen solubility in air saturated fresh water mg L Appendix 61 T C cs T 1 2 3 A 5 6 7 8 9 1 0 0 14 64 60 55 51 47 43 39 35 EY 27 23 1 23 19 15 10 06 03 99 95 91 87 83 2 13 83 79 75 71 68 64 60 56 52 49 45 3 45 41 38 34 30 27 23 20 16 12 09 4 09 05 02 98 95 92 88 85 81 78 75 5 12 75 71 68 65 61 58 55 52 48 45 42 6 42 39 36 32 29 26 23 20 17 14 11 7 11 08 05 02 99 96 93 90 87 84 81 8 11 81 78 75 72 69 67 64 61 58 55 53 9 53 50 47 44 42 39 36 33 31 28 25 10 25 23 20 18 15 12 10 07 05 02 99 11 10 99 97 94 92 89 87 84 82 79 77 75 12 75 72 70 67 65 63 60 58 55 53 51 13 51 48 46 44 41 39 37 35 32 30 28 14 28 26 23 21 19 17 15 12 10 08 06 15 06 04 02 99 97 95 93 91 89 87 85 16 ER 85 83 81 70 76 74 72 70 68 66 64 17 64 62 60 58 56 54 53 51 49 47 45 18 45 43 41 39 37 35 33 31 30 28 26 19 26 24 22 20 19 17 15 13 11 09 08 R ES 06 04 02 01 99 97 95 94 92 90 21 8 90 88 87 85 83 82 80 78 76 75 73 22 73 71 70 68 66 65 63 62 60 58 57 23 57 55 53 52 50 49 47 46 44 42 41 24 41 39 38 36 35 33 32 30 28 27 25 25 25 24 22 21 19 18 16 15 14 12 11 26 11 09 08 06 05 03 02 00 99 98 96 27 7 96 95 93 92 90 89 88 86 85 83 82 28 82 81 79 78 77 75 74 73 71 70 69 29 69 67 66 65 63 62 61 59 58 57 55 30 55 54 53 51 50 49 48 46 45 44 42 31 42 41 40 39 37 36 35 34 32 31 30 32 30 29 28 26 25
57. Phase o oO Amplitude oO pO kPa 120 63 LO pO kPa Jj 0 00 HI pO kPa J 20 96 Chamber oxygen 4 Mov Avg lpoint x Lock LO Lock HI ILTR Mov Avg lpoint YV Lock LO Lock HI Enable pO kPa 0 00 LO Temp C Phase 20 00 D HI Temp C Amplitude 20 00 o Analog Comp Input V Temp C 4 87 97 35 LO Temp C J 0 00 LO input v 0 00 y HI Temp C J 100 00 HI input v 1j5 00 y Fiber Optic ON Oxygen graph Save as Set temp C 20 00 ce r was as y 10 00 10 00 joo 10 00 1 14 22 39 Page 30 of 50 L oii o LOLIGOSYSTEMS COM 5 2 13 Chamber oxygen field 1 4 5 2 13 1 Mov Avg button Use this button to select if the chamber oxygen input signal should be averaged or not The default value is 1 point which means no averaging of the input If any other value is chosen the chamber oxygen input will be averaged and the averaged values will be used for all subsequent computations screen and file outputs Use moving average if you wish to smoothen the chamber oxygen signal Please note No calibrated values will be displayed either on screen or in the data file before sufficient samples have been collected to compute the moving average E g if the moving average is set to 10 points and all four
58. age 30 A checkmark appears named Analog Comp 5 2 14 1 Analog Comp Use this checkmark to enable disable the compensation of the fiber optic oxygen signal with the analog measured temperature If the temperature changes with 0 2 C the new temperature will be send to the fiber optic instrument Page 32 of 50 5 3 Setup experiment 5 3 1 Screen 24 l AutoResp 4 Help File name Setup Flush period s 240 O solubility mgO 0 4855 Wait period s jo WMkPa These are the notes about the experiment General about Screen 24 On Screen 24 the intermittent respirometry experiment is configured Measure period s 1300 y Chamber 1 Resp vol I 1 0000 Wet weight g Jj 100 Chamber 2 Wet weight g 1j 100 y Resp vol I H 1 0000 y Chamber 3 Resp vol I t1 1 0000 Oxygen control Setpoint kPa Jj 20 50 Wet weight g J 100 i s Hysteresis kPa Jj 0 10 Chamber 4 Resp vol I 1 1 0000 Wet weight g 13100 Temperature control Setpoint C 1j15 00 y Hysteresis C L oii o LOLIGOSYSTEMS COM Jj 0 10 Cool Hypoxic O Hyperoxic Heat O Start experiment 5 3 2 File name button Use this button to change the path and filename of the data file and raw data folder The path specifies the location of the main data file and the folder containing raw data from the experiment T
59. and completely PC controlled fiber optic oxygen meter The data evaluation is PC supported as well The OXY 4 was specially developed for small fiber optic oxygen sensors flow through cells and integrated sensor systems It is based on a novel technology which creates very stable internally referenced measured values This allows a more flexible use of oxygen sensors in many different fields of interest Optical oxygen sensors also called optodes have important advantages over common Clark type electrodes e They are small e They do not consume oxygen e Their signal does not depend on the flow rate of the sample e They have an excellent long term stability e They can be physically divided from the measuring system which means a contactless measurement e They can be autoclaved and y sterilized Therefore they are ideally suited for the examination of small sample volumes long term measurements in difficult sam ples and for biotechnological applications A set of different oxygen minisensors flow through cells and integrated sensor systems is available to make sure you have the sensor which is ideally suited to your application Please feel free to contact our service team to find the best solution for your application Your PreSens Team S Safety Guidelines a 2 Safety Guidelines PLEASE READ THESE INSTRUCTIONS CAREFULLY BEFORE WORKING WITH THIS INSTRUMENT This device has left our works after careful testing of all function
60. are shown The first field shows the chosen units which can be altered by the unit button To distinguish the different oxygen signal from each chamber every oxygen signal has its own color Oxygen signal Color Chamber 1 Light Blue Chamber 2 Green Chamber 3 Dark blue Chamber 4 Orange Ambient Lila 5 2 11 1 Save button Use this button to save the data displayed in the Oxygen graph Furthermore the temperature will be saved When pressed the user is prompted to enter a file name and path see Screen 22 The path is by default the installation directory of AutoResp4 AutoResp4Data The current time and date xls is suggested as a filename in the prompt Both the path and filename can be altered by choice The system suggests two file extensions xls and txt The xls extension will create a tabulator separated text file which is readable by Microsoft Excel The txt extension will create a semicolon separated text file which is readable by any Microsoft Windows PC 5 2 11 2 Unit button Use this button to convert the displayed oxygen partial pressure values on the chamber oxygen graph into Torr oxygen saturation or air saturation 5 2 11 3 Play button Use this button to start stop data from being graphed When the play button is ON new data will be shown on the oxygen graph and the Unit button is enabled When the play button is OFF the Unit button is disabled Furthermore a scroll bar is sho
61. ase angles are constant the variation of the phase angles should be smaller than 0 05 and press the CAL 0 button to store the 0 air saturation calibration values at the adjusted temperature A message window opens and informs you that you will program all channels with the actual calibration values Click the OK button to store the new calibration data 4 Rinse the sensor membranes gently with water after calibration to remove sulfite remaining on the surface Calibration 41 Calibrate Single Channels 1 Place the calibration solution 100 cal 100 air saturated water or water vapor saturated air into the glass vessel To minimize the response time slightly stir the solution Please ensure that the cal 100 solution completely covers the sensor foil Measurement Calibration Logging All channels XX 2nd cal point E vdd CHANNEL eee 1013 e 4 temperature of cal standard E 200 temperature of cal100 standard 20 0 Oxygen Sam sat 99 80 bM TE 1013 HEG 25 19 Temp cal Amplitude 44285 Temp cali 00 2 Wait about 3 minutes until the phase angle is constant the variation of the phase angle should be smaller than 0 05 and click the cal 100 button to store the 100 air sat calibration value at the adjusted temperature A message window opens and informs you that you will change the last calibration values Click the OK button to store the new calibration data Repeat this proc
62. asurement amp Automation by following the folder hierarchy shown on Screen 15 Page 13 of 50 Screen 16 LOLIGOSYSTEMS COM C NI USB 6215 Dev1 Measurement amp Automation Explorer ERE X File Edit View Tools Help Configuration Ay E Properties X Delete Z Self Test SM Test Panels fJ ResetDevice f Create Task Ga Configure TEDS Hide Help Bg e My System amp Bil Data Neighborhood Name Value Geax El Al amp gg Devices and Interfaces Dea Number OxE669F2 NI NIUSE 6215 Devi DAGmx Device gg PXI PXI System Unidentified E 5 B3 Scales Basics amp aj Software amp e Remote Systems What do you want to do Run the NI DAGmx Test Panels Remove the device View or change device properties T amp J Mj screen 21 bmp Paint NI USB 6215 Devi 2X wa aur TERK SS seam Make sure the NI USB 6215 is installed as device name Dev1 If so the DAQ 4 instrument is now properly correct installed and ready for use with AutoResp 4 If device name not should be dev1 see 6 1 1 Name is not Devi on page 41 20 Remove CD labelled AutoResp 4 Page 14 of 50 L oii o LOLIGOSYSTEMS COM 3 4 OXY 4 fiber optic oxygen instruments 3 4 1 Front side CH1 Fiber optic oxygen input for chamber 1 CH2 Fiber optic oxygen input for chamber 2 CH3 Fiber optic oxygen input for chamber 3 CH4
63. ation on the vapor pressure is negligible small as shown in Table 12 5 Table 12 5 Variation of solution vapor pressure pw with salt concentration Vapor pressure of solution torr Frere 9 3 6 45 4 5 20 The dependence of oxygen solubility on salt concentration can also be obtained from equation 21 except that now values calculated from either equation 27 or 28 have to be used for calculation of the Bunsen absorption coefficient Equation 27 differs from equation 22 by an additional forth degree polynomial term for chlorinity 10 a a b 0 c 0 e d 0 e 6 CI p q O r 0 s 0 t 6 27 where 0 is the temperature in C a e are the coefficients used in equation 22 and p t are new constants given in Table 12 6 The values of these new constants are obtained by fitting the polynomial to experimental data in the ranges 0 lt 0 x 30 C and 0 lt CI lt 20 To obtain an oxygen solubility from the Bunsen absorption coeffi cient the same procedure as described previously is used s equation 24 page 60 An alternative equation to compensate the Bunsen absorption coefficient by the salt concentration displays equa tion 28 10 azep A BecuntsD T 10r P 24R nr s T 28 where T is the temperature in Kelvin and A D and P S are the coefficients given in Table 12 6 They are based on measurements for 273 1 x T lt 308 18 K and 0 lt CI lt 30 and is therefore more extensive than equation 27
64. bending or by intensity changes caused by changes in the geometry of the sensor e The decay time is to a great extent independent of the concentration of the indicator in the sensitive layer 2 photobleaching and leaching of the indicator dye has no influence on the measuring signal e The decay time is not influenced by variations in the optical properties of the sample including turbidity re fractive index and coloration 12 1 5 Literature If you want to find out more about this subject we recommend the following publications e Wolfbeis O S Ed Fiber Optic Chemical Sensors and Biosensors Vol 1 amp 2 CRC Boca Raton 1991 e Klimant I Wolfbeis O S Oxygen Sensitive Luminescent Materials Based on Silicone Soluble Ruthe nium Diimine Complexes Anal Chem 67 3160 3166 1995 e Klimant I K hl M Glud R N Holst G Optical measurement of oxygen and temperature in microscale strategies and biological applications Sensors and Actuators B 38 39 29 37 1997 e Holst G Glud R N K hl M Klimant A microoptode array for fine scale measurement of oxygen dis tribution Sensors and Actuators B 38 39 122 129 1997 e Klimant l Meyer V K hl M Fiber optic oxygen microsensors a new tool in aquatic biology Limnol Oceanogr 40 1159 1165 1995 e Klimant l Ruckruh F Liebsch G Stangelmayer A Wolfbeis O S Fast Response Oxygen Microsensors Based on Novel Soluble Ormosil Glasses Mikrochim Acta
65. brate the optical sensors One is the use of two calibration standards the other one is to type in known constants The first way needs two calibration standards Their preparation is described in chapter 7 3 1 The second possibility is described in chapter 7 3 2 7 3 1 Calibration with two calibration standards For the calibration with current values you need calibration standards and your mounted sensor Calibration of the minisensors is performed using conventional two point calibration in oxygen free water cal 0 and water vapor saturated air or air saturated water cal 100 In the sub window Calibration cal 100 is denoted as 2 cal point 7 3 1 1 Preparation of the Calibration Standards Preparation of calibration solution 0 oxygen free water 1 Add 1 g sodium sulfite Na2SO to the vessel and label it cal 0 2 Dissolve Na SOsin 100 mL water The water becomes oxygen free due to a chemical reaction of oxygen with Na2SOs Additional oxygen diffusing from air into the water is eliminated removed by surplus of Na2SOs 3 Close the vessel with a screw top and shake it for approximately one minute to dissolve NaSO and to ensure that wa ter the is oxygen free Close the vessel after calibration with a screw top to minimize oxygen contamination To prepare oxygen free water you also can use sodium dithionit NazS2O4 The shelf life of cal 0 is about 24 hours pro vided that the vessel has been closed with the screw top Pr
66. button The oxygen content is displayed in the chosen unit the temperature in C The raw values the phase angle in degrees and the sensor amplitude in mV are also displayed see Appendix for description of phase and amplitude Software 24 Measurement Carton Loong Al chanet Channels Graphical window legend If Autoscale is Off click here to change values After that click some where inside the graphi cal window to make changes active Unzoom button Clear graph button Scale button Data choice button 6 3 5 Graphical and numerical display of the respective channel The measuring values and all raw values are displayed for each channel separately By clicking on the respective channel the actual measured oxygen concentration is shown in the unit you have adjusted in the menu bar measurement The raw values amplitude and phase value can be displayed by clicking on Raw values on in the Channel settings on the right side or they can be hidden by clicking on off Channel 1 settings Clear graph auto scale Channel settings In the channel settings the user is able to configure each channel separately and to optimize the instrument performance 1 Raw values The raw values the phase angle in degrees and the sensor amplitude in mV can be displayed hidden by clicking the On Off button If the raw values are not displayed the signal amplitud
67. calibration screen as it looks if chamber oxygen input is a 0 5 VDC analog signal e g from an OXY REG or some other O instrument On this Screen a common sample frequency for all inputs can be chosen and the inputs can be calibrated from voltages into oxygen partial pressures and temperature This screen can also be used to do a quick oxygen or temperature measurement Please note The calibration values is not stored until an experiment has been started That means the calibration values are NOT stored when the Next gt button is pressed Barometic pressure hPa Chamber oxygen 1 Mov Avg 1 point x Lock LO Lock HI Hee Input V 3 12 LO input V 130 00 y HI input v 15 00 y 1013 pO kPa 113 10 0 00 7 HI pO kPa 1320 96 y pO sat kPa 20 859 J Chamber oxygen 2 Mov Avg ipoint vV Input V 2 33 LO input v 30 00 y Hi input v 115 00 y Fiber Optic OFF C Oxygen graph awa ead 9 77 25 LO poz kPa Jj 0 00 22 5 HI pO kPa Jj 20 96 Chamber oxygen 3 Mov Avg lpoint x Lock LO Lock HI Ati Input V 2 11 LO input V 130 00 y Hl input e 115 00 y pO kPa 8 95 LO po kPa Jooo HI pO kPa 20 96 Chamber oxygen 4 Mov Avg lpoint x Input V 1 90 LO input v 79 00 HI input V Jj 5 00 pO kPa 17 95 LO pO kPa 5 J0 00 HI pO kPa 1320 96 Ambient oxygen Mov Avg
68. d label it cal 0 2 Dissolve Na SOs in 100 mL water The water becomes oxygen free due to a chemical reaction of oxygen with Na2SOs Additional oxygen diffusing from air into the water is removed by surplus of Na2SOs 3 Close the vessel with a screw top and shake it for approximately one minute to dissolve NaSO and to ensure that the water is oxygen free Close the vessel after calibration with a screw top to minimize oxygen contamination To prepare oxygen free water you also can use sodium dithionit Na2S2O4 The shelf life of cal 0 is about 24 hours pro vided that the vessel has been closed with the screw top Preparation of calibration standard cal 100 air saturated water 1 Add 100 mL water to a suitable vessel and label it cal 100 2 To obtain air saturated water blow air into the water using an air pump with a glass frit airstone creating a multitude of small air bubbles while stirring the solution 3 After 20 minutes switch of the air pump and stir the solution for further 10 minutes to ensure that the water is not supersaturated 7 2 1 2 Mounting the Oxygen Sensitive Minisensors 1 Remove the flow through cell oxygen sensor carefully from the protective cover The oxygen sensitive material is im mobilized to a glass tube which is located in a T connector with two female Luer Lock adapters The glass tube is tightened with a viton seal and two male Luer Lock connectors fix the glass tube in the T connector On requ
69. e OXY 4 we only consider ourselves responsible for safety and performance of the device if e the device is strictly used according to the instruction manual and the safety guidelines e the electrical installation of the respective room corresponds to the DIN IEC VDE standards The OXY 4 and the sensors must not be used in vivo examinations on humans The OXY 4 and the sensors must not be used for human diagnostic or therapeutical purposes L __escriptionofthe OXY 10 Device 8 3 Description of the OXY 4 Device 3 1 Instrument The OXY 4 is a 4 channel oxygen meter for use with fiber optic oxygen minisensors based on a 2mm polymer optical fiber POF The OXY 4 system detects oxygen the oxygen partial pres sure in both solutions dissolved oxygen as well as in the gaseous phase For operation a PC Notebook with RS 232 inter face is required The OXY 4 is controlled using a comfortable software which also saves and visual izes the measured values OXY 4 does not contain temperature sensors temperature changes during the measurement are not compensated by the software 3 2 Front Panel of the OXY 4 Device ELEMENT DESCRIPTION FUNCTION CH1 CH4 SMA fiber connector Connect the fiber optic oxygen minisensor here Line Blue Control on instrument on LED off instrument off Description of the OXY 10 Device 4 3 3 Rear Panel of the OXY 4 Device The electrical specifications of all rear panel connectors are g
70. e fiber plug and connect it to one of the SMA plugs of the OXY 4 device The safety nut must be carefully attached while turning slightly clockwise 7 1 1 3 Performing the calibration 1 Switch on the OXY 4 and the PC software following chapter 6 2 Goto window Calibration and enter the actual atmospheric pressure in hPa and the temperature of the calibration standards cal 0 and cal 100 The atmospheric pressure of the calibration is needed to convert the oxygen unit air saturation into partial pressure units hPa Torr or concentration units mg L pmol L Please ensure that there are no temperature changes during the calibration of the oxygen sensor Besides it must be ensured that the temperature during later measurement is constant and already known However the temperatures during the measurement and the calibration process are allowed to be different 3 Select the channel to be calibrated in the section Channel of sub window SINGLE CHANNEL or switch to the sub window ALL CHANNNELS Calibration 30 Calibrate All Channels Place the calibration standard 100 cal 100 containing wet cotton wool underneath the oxygen minisensors The vessel with the label cal 100 has to be closed with the screw top containing the holes Insert the plastic fibers carefully through the holes without touching the oxygen sensitive spots until they are about 3 cm deep inside the vessel Make sure that the plastic fibers with their sensor spots cannot
71. e is shown via a bar the longer the bar the higher the signal 2 Signal LED intensity Note After changing the intensity any sensor calibration for the changed channel has to be repeated With the current of the LED you can adjust the amount of light illuminating the sensor spot The LED current can be set manually between 10 and 100 If you increase the LED current the signal amplitude increases since a higher light density illuminates the sensor spot The channel settings can be adjusted for the respective chosen channel by clicking set one However it is also possible to set all channels with the same value by clicking set all Please note By increasing the light intensity you increase the amplitude of the oxygen microsensor This leads to smoother phase signals However increasing the light intensity can in crease photobleaching which decreases the shelf life of your sensor Software 26 Four windows are used to display the data of each channel separately The windows named channel 1 to channel 4 show the data of the respective channel The oxygen unit is the same as specified in the window Measurement at the beginning of the measurement It is not possible to change the unit during the measurement The last measured oxygen value is dis played in the marked field on the left The scale of the y axis and the starting point of the x axis can be set to different modes by clicking on the Scale button Two options are implem
72. e minute to dissolve Na2SOs and to ensure that the water is oxygen free Close the vessel after calibration with a screw top to minimize oxygen contamination The shelf life of cal 0 is about 24 hours provided that the vessel has been closed with the screw top Preparation of calibration standard cal 100 water vapor saturated air 1 Place wet cotton wool in the vessel with the label cal 100 2 Drill a hole for inserting the minisensor in the screw top and close the vessel If you want to calibrate several minisen sors simultaneously drill as many holes as there are minisensors 3 Wait about 2 minutes to ensure that the air is water vapor saturated Alternatively Preparation of calibration standard cal 100 air saturated water 1 Add 100 mL water to a suitable vessel and label it cal 100 2 To obtain air saturated water blow air into the water using an air pump with a glass frit airstone creating a multitude of small air bubbles while stirring the solution 3 After 20 minutes switch off the air pump and stir the solution for further 10 minutes to ensure that water is not super saturated 7 1 1 2 Mounting the Oxygen Sensitive Minisensors 1 Remove the oxygen sensor carefully from the protective cover 2 Carefully remove the protective plastic cap covering the oxygen sensitive sensor spot 3 Fixthe oxygen sensor with a clip to a laboratory support or a similar stable construction 4 Remove the protective cap from the mal
73. edure for all other channels by typing in the respective channel into the Channel Window to record the 100 calibration value cal 100 3 To record the second calibration value oxygen free water cal 0 remove the cal 100 solution from the vessel and fill it with the calibration standard 0 cal 0 To minimize the response time slightly stir the solution Please note Vigorous stirring can lead to an oxygen contamination of the cal 0 solution Wait about 3 minutes until the phase angle is constant the variation of the phase angle should be smaller than 0 05 and press the CAL 0 button to store the 0 air sat calibration value at the adjusted temperature A message window opens and informs you that you will change the last calibration values Click the OK button to store the new calibration data Repeat this procedure for all other channels by typing in the respective channel into the Channel Window to record the oxygen free calibration value cal 0 4 Rinse the sensor membranes gently with water after calibration to remove sulfite remaining on the surface Calibration 42 7 3 2 User defined calibration If temperature and phase angle of a former calibration of the same sensor are known the Calibration with constant values is possible 1 2 3 Switch on the OXY 4 and the PC software following chapter 6 Go to the window Calibration Select the channel to be calibrated in the section Channel of sub window Single
74. eneral power series to the values in Table 12 2 A fourth degree polynomial fit can be chosen yielding equation 22 10a a b 0 c 0 d 0 e 0 22 where 0 is the temperature in C and a e the coefficients calculated by standard curve fitting procedures as given in Figure 12 8 50 o 0 10 48 998 1 335 0 2 755 10 7 9 3 220 10 0 1 598 10 0 a 48 998 b 1 335 40 c 2 755 10 d 3 220 107 e 1 598 10 30 20 0 5 10 15 2 0 25 30 35 40 45 50 6 C Figure 12 8 Variation of Bunsen absorption coefficient a 0 with temperature Ff is the square of the correlation coefficient The other form of equation to describe the variation of a with temperature can be derived from a thermodynami cal correlation and gives an equation of the form Appendix 60 Inl0 a 24B InT C 23 where A B and C are constants and T is the temperature in K For oxygen dissolved in water we find by fitting the equation to the values of a in Table 12 2 that A 8 553 10 B 2 378 10 and C 1 608 10 Values of a calculated from eqns 22 and 23 for the same temperature agree within 0 5 96 The Bunsen absorption coefficient however is not a very practical measure Values of a 0 have therefore to be converted to mg L and the method for doing this is best illustrated by an example Example Calculation of the oxygen content Cs patm 9 in air saturated water at a temperature 0 of 20 C Equati
75. ented 1 Autoscale On The software determines the minimum and maximum values to display all data 2 Autoscale Off The maximum and minimum value of y axis and the minimum of the x axis can be defined by the user By clicking on the current value the minimum and or maximum value can be changed The Clear graph button is used to clear the graphical window from all former data 6 4 Data Handling In the head of the ASCII file you find Line 1 time and date when the measurement has started software version Line 2 number of measured channel Line 3 5 description of the measurement if a content is added to the File description section max 3 lines Line 6 10 Identification information of the hardware only useful for service Line 12 19 Set parameters of the hardware only useful for service Line 21 26 System settings only useful for service Line 28 33 Information about the calibration which was used for the measurement useful if a mistake during calibration is supposed Line 35 37 Information about the firmware only useful for service Line 39 Pressure at which calibration was performed Line 40 xx The following rows separated by semicolons list the measuring data The first two rows con tain the date and time the third the log time in minutes the fourth the oxygen content in the chosen unit The raw data phase angle in and the amplitude in mV are stored in the fifth
76. eparation of calibration standard 100 water vapor saturated air 1 Place wet cotton wool in the vessel and label it cal 100 2 Drill two holes for inserting the minisensor and the temperature sensor in the screw top and close the vessel with it 3 Wait about 2 minutes to ensure that air is water vapor saturated Preparation of calibration solution 100 air saturated water 1 Add 100 mL water to a suitable vessel and label it cal 100 2 To obtain air saturated water blow air into the water using an air pump with a glass frit airstone creating a multitude of small air bubbles while stirring the solution 3 After 20 minutes switch off the air pump and stir the solution for further 10 minutes to ensure that the water is not supersaturated Calibration 39 7 3 1 2 Mounting the Oxygen Sensitive Minisensors 1 Remove the oxygen sensitive foil carefully from the protective cover 2 Glue small spots of the oxygen sensitive foil into the desired glass vessel using transparent silicone The transpar ent silicone can be purchased from PreSens left Oxygen sensitive foil glued on the inner surface of a glass vial right Glass vial with integrated oxygen sensitive foil in appropriate adapter Please note Be sure to glue the sensor spots onto your vessel with the proper side The sensor support polyester foil or glass on which the sensor is spotted identifiable by its faint reflection is glued to the vessel while the sensor itse
77. erane E 23 3 A convenient fitting function is given by the Campbell equation 20 Pw T ew A C ln T 20 where T is the temperature in Kelvin and A B and C constants given in Figure 12 7 120 T py T exp 52 57 6690 9 T 4 681 InT R 1 100 z t UH A 52 57 iam 7 B 6690 9 e 60 T C 4 681 E Qo 40 20 0 T T T T T T T T T T T T l T l T T T T T 272 277 282 287 292 297 302 307 312 317 322 T K Figure 12 7 Variation of water vapor pressure with temperature F is the square of the correlation coefficient Appendix 59 12 4 2 Bunsen Absorption Coefficient The solubility of oxygen in water is temperature dependent and can be described using the Bunsen absorption coefficient a 0 and the oxygen partial pressure p Oz according to equation 21 With increasing temperature the solubility of oxygen in water decreases p O py T T N c p 0 0 21 Cs p 8 temperature dependent solubility of oxygen in water given in cm O2 cm p O2 oxygen partial pressure PN standard pressure 1013 mbar o 0 Bunsen absorption coefficient given in cm Oz cm Table 12 2 Variation of Bunsen absorption coefficient a 0 with temperature ee 9 p 20 The data in Table 12 2 can be described by two forms of equations 30 The first form of equation to describe the temperature dependent variation of the Bunsen absorption coefficient a 0 is obtained by fitting a g
78. es during the measurement and the calibration process are allowed to be different 3 Select the channel to be calibrated in the section Channel of sub window SINGLE CHANNEL or switch to the sub window ALL CHANNNELS Calibrate All Channels 1 Place the calibration solution 100 cal 100 air saturated water or water vapor saturated air into the glass ves sels To minimize the response time slightly stir the solution Please ensure that the cal 100 solution completely covers the sensor foils Measurement Calibration Logging All channels 2nd cal paint 100 00 ek pressure 1013 hPa temperature of call standard 20 0 AE temperature of cal100 standard amp 200 2 Wait about 3 minutes until the phase angles are constant the variation of the phase angles should be smaller than 0 05 and click the CAL 100 button to store the 100 air sat calibration values at the adjusted temperature A message window opens and informs you that you will program all channels with the actual calibration values Click the OK button to store the new calibration data 3 Torecord the second calibration values oxygen free water cal 0 remove the cal 100 solution from the vessels and fill them with the calibration standard 0 cal 0 To minimize the response time slightly stir the solution Please note Vigorous stirring can lead to an oxygen contamination of the cal 0 solution Wait about 3 minutes until the ph
79. esatutah M Scene ip ae i aar satura 5005 cep 000 MCI 15 j Py 100 M in ppm mg L hub Voca e satt M co ppm co mg LJ Pam Pw CD air saturation 9 2095 aT 1999 MO 2 ao Py 100 M in umol L 1000 c mol L co mg L c mg L 31 25 o IH Co mg L M 0 o mg L 17 PP Su Pam Pw D 96 sir saturation 09 T 1000000 Py 100 V Patm actual atmospheric pressure pn standard pressure 1013 mbar 0 2095 volume content of oxygen in air pw T vapor pressure of water at temperature T given in Kelvin a T Bunsen absorption coefficient at temperature T given in cm O2 cm M Oz molecular mass of oxygen 32 g mol Vu molar volume 22 414 L mol 57 Appendix 58 12 4 Temperature Dependent Constants Affecting the Oxygen Content 12 4 1 Water Vapor Pressure As shown in equation 12 17 the water vapor pressure py influences the oxygen partial pressure of air saturated water and water vapor saturated air Oxygen partial pressure in dry air p O Pam 0 2095 18 p Oz oxygen partial pressure in dry air at a barometric pressure Patm 0 2095 volume content of oxygen in air Oxygen partial pressure in air saturated water and water vapor saturated air p O 7 Pam Pw T 0 2095 19 Temperature variations strongly affect water vapor pressure and thus influence the oxygen partial pressure as shown in equation 19 Table 12 1 Variation of water apor pressure pw T with p
80. est the seal can also be out of silicone Don t remove the two male Luer Lock adapters from the T connector You may loose the viton seal and the glass tube may be dislocated male Luer Lock connector containing a 2 mm POF located at the glass tube male Luer Lock connector male Luer Lock connector viton seal glass tube with viton seal integrated oxygen sensor 2 Fix the male Luer Lock adapter with the integrated 2 mm POF polymer optical fiber to the Luer T connector and ensure that the fiber is located close to the glass tube Calibration 34 Fix the flow through cell with a clip to a laboratory support or a similar stable construction Connect the two male Luer Lock connectors with the tubings of your flow through system Remove the protective cap from the male fiber plug and connect it to the SMA plug of the OXY 4 device The safety nut must be carefully attached while turning slightly clockwise 7 2 1 3 Performing the calibration 1 Switch on the OXY 4 and the PC software following chapter 6 2 Go to window Calibration and enter the actual atmospheric pressure in hPa and the temperature of the calibration standards cal 0 and cal 100 The atmospheric pressure of the calibration is needed to convert the oxygen unit air saturation into partial pressure units hPa Torr or concentration units mg L pmol L Please ensure that there are no temperature changes during the calibration of the oxygen sensor Besides it m
81. evel oxygen calibration of the chamber oxygen input Please note The field is limited to values between 0 00 kPa to 60 00 kPa 5 2 8 7 Lock HI button This button can be used to copy the actual value from the Input V indicator to the HI input V field instead of manually reading and retyping the value Page 25 of 50 L oii 0 5 2 8 8 HI input V LOLIGOSYSTEMS COM Use this field to enter the high input voltage from the Input V indicator during the chamber oxygen calibration Please note The field is limited to values between OV to 5V 5 2 8 9 HI pO kPa Use this field to enter the oxygen pressure during the high level oxygen calibration of the chamber oxygen input Please note The field is limited to values between 0 00 kPa to 60 00 kPa 5 2 9 Ambient oxygen field 5 2 9 1 Mov Avg button Use this button to select if the ambient oxygen input signal should be averaged or not The default value is 1 point which means no averaging of the input If any other value is chosen the ambient oxygen input will be averaged and the averaged values will be used for all subsequent computations screen and file outputs Use moving average if you wish to smoothen the ambient oxygen signal Please note No calibrated values will be displayed either on screen or in the data file before sufficient samples have been collected to compute the moving average F g if the moving average is set to 30 points and the sample rate is set to 10 Hz
82. f 50 LOLIGOSYSTEMS COM 8 4 OXY 4 Instruction manual The instruction manual for the OXY 4 fiber optic instrument from PreSens manual is included as an individual document Please use this manual for further information about the OXY 4 fiber optic oxygen instrument and the OXY 4 software Please look into the DAQ PAC F4S package Page 50 of 50 L oii o LOLIGOSYSTEMS COM Instruction Manual OXY 4 4 Channel Fiber Optic Oxygen Meter L oii o LOLIGOSYSTEMS COM Instruction Manual OXY 4 Software Version OXY4v2_11FB May 2005 Table of contents Table of Contents 1 Preface iie iacuietiiir dietas reri sede se ia Mo DEA dS coria Od T 1 2 Safety Guidelines iei enin ede eine iteids eee le perle eerie 2 3 Description of the OXY 4 Device eeeeeeeeeeeeeeeeeee ee eenee enne nn nnn nn nnne natnra nnne sa inns ns snas etna sites sinn sse nn sana 3 3 1 EI Dru e E E E A E A M 3 3 2 Front Panel of the OXY 4 Device cssecsseeeseseeeeeeeeseeeseesaseeesseeeaseeeesaeesseeesseeeasaeeeseeeaseeseseesaseeseeeas 3 3 3 Rear Panel of the OXY 4 Device uriieie tecti redirent racconti ent catene cierta nta aa Bap seins mud muds 4 Required Basic Equipment eeseeeeee esses esee eee eeenn enne ennn nnn ntna nnne nani nenas sss naai naiean napainen ada dianian 5 Planar Oxygen Minisensors eeeeeeieeeeeeeeee eene eene nennen nnns nnne na nnn
83. f Labview Run Time Engine Now the Software is properly installed and you will find a link in your Start menu Software 20 6 2 Starting OXY 4 and its Software 1 Connect the OXY 4 via the supplied serial cable to a serial port of your computer Tighten the cable with the screws on your computer and on the OXY 4 2 Connect the power supply 3 Please close all other applications as they may interfere with the software Start the program OXY4v2 09 in the Start menu The following information window appears 47 mm z initialization Fa Please connect the instrument to the PC and turn the power switch on Next please select the proper serial port number Serial Port 4 Choose the right com port with a left mouse click on Serial Port Then press OK The following window appears and all 4 channels are initialized Loading data from channel 1 If the wrong com port is adjusted the software gives an error message Please change the com port and press OK 6 3 Function and Description of the OXY 4 Software The window shown below is displayed after starting the software OXY 4 software The program has 5 main sections 1 Measurement Calibration Logging All Channels Graphical and numerical display of all channels a FF oO DN Graphical and numerical display of the respective channel o Ax E e 7 AO 4v2_11 DATE TIME SAMPLING CHANNEL RATE TEMPERATURE locked mj locked mj ALL CHAN
84. f call standard temperature of cal100 standard Oxygen Pressure eee 200 Amplitude nu 1265 Temp cal1O0 20 p 95 100 00905 a Phase 56 09 27 00 Figure 7 2 Screenshot of the sub window ALL CHANNELS of the window Calibration Calibration 29 7 1 Calibration of Oxygen Dipping Probe DP PSt3 There are two ways to calibrate the optical sensors One is the use of two calibration standards the other one is to type in known constants The first way needs two calibration standards Their preparation is described in chapter 7 1 1 The second possibility is described in chapter 7 1 2 7 1 1 Calibration with two Calibration Standards For the calibration with current values you need calibration standards and your mounted sensor 7 1 1 1 Preparation of the Calibration Standards Calibration of oxygen minisensors is performed using a conventional two point calibration in oxygen free water cal 0 and water vapor saturated air or air saturated water cal 100 In the sub window Calibration constants cal 100 is denoted as 2 cal point Preparation of calibration standard cal 0 oxygen free water 1 Add 1 g sodium sulfite Na SO to a vessel and label it cal 0 2 Dissolve NazSO3in 100 mL water The water becomes oxygen free due to a chemical reaction of oxygen with Na2SOs Additional oxygen diffusing from air into the water is removed by surplus of Na2SOs 3 Close the vessel with a screw top and shake it for approximately on
85. g usi RE ee e E a ar E ave e aeo 22 6 3 4 All Channels Graphical and numerical of all channel eene 23 6 3 5 Graphical and numerical display of the respective channel 25 6 4 Data Handling me a MH 26 7 Calibration EC M 28 7 1 Calibration of Oxygen Dipping Probe DP PSI3 eeseeieesseseeeeeee nennen nnne nnns nna nnnan 29 7 1 1 Calibration with two Calibration Standards sseeeeeeeneeneeeneen enne 29 7 1 2 User d fined calibration 2 eer tet ete eet tte Tee tede ists 32 7 2 Calibration of Flow Through Cell FTC PSI3 esses eese nennen tnnt annus 33 7 2 1 Calibration with two calibration standards ssessseseeeeeeeenenennenen nennen 33 1 22 2 Jser defined calibration teen e eoe eese nene 37 7 3 Calibration of Planar Oxygen Sensitive Foils PSt3 Integrated in Glass Vials 38 7 3 1 Calibration with two calibration standards sssseeeeenenenenen nennen 38 7 3 2 Aser defined calibration t eR ee ERE A A PEOR 42 8 Measurement with Oxygen Sensor PSt3 eeeeseeseeeeeeeeeee eene n nene na nnn ennt natns nnn nantes nnn 43 8 1 Starting the measurement 4 essei esses eeeeeeeen ene nnn nnns anten nnnm annnm ase tn ase tnn sita natns sinn asa sn sna 43 8 1 1 Scarining all Ghantnels 2
86. gle in absence of oxygen o Figure 12 11 and the Stern Volmer constant Ksv starting slope in Figure 12 12 according to equation 4 are temperature dependent o decreases with increasing temperature while Ksy increases with increasing tempera ture Appendix 64 o2 I Nn nN phase angle N 10 0 20 40 60 80 100 120 140 160 180 200 220 240 air saturation Figure 12 11 Effect of the temperature on the phase angle at different oxygen contents given in air saturation 6 40 C 30 C 20 10 C 5 2 C 2 R 4 gt 2 a 0 20 40 60 80 100 120 140 160 180 200 220 240 260 air saturation 46 Figure 12 12 Effect of the temperature on the Stern Volmer constant Figure 12 13 displays the temperature dependence of p and Ksy From Figure 12 13 decreases in doof about 0 09 can be calculated by increasing the temperature by 1 K On the other hand the Stern Volmer constant Ksv increases about 5 0 10 96 air sat by increasing the temperature by 1 K Appendix 65 61 9 y 62 14 0 08915 x 0 07 0 068 0 066 0 064 0 062 0 06 0 058 0 056 0 054 y 0 04899 4 965 107 x 0 052 R 0 99948 R 0 99914 61 4 60 9 60 4 o 59 9 59 4 Koy air saturation 58 9 58 4 0 05 2 4 10 2 20 4 30 4 40 3 temperature C Figure 12 13 Effect of the temperature on o and the Stern Volmer constant Ksv Consequently variations
87. greement s I do not accept the License Agreement s v 7 Then click Next On Screen 5 a summary is given of the products to be installed Screen 5 Start Installation Review the following summary before continuing Adding or Changing AutoRespl 4 Files NI DAQms 8 5 NI LabVIEW Run Time Engine 7 1 1 NI Measurement amp Automation Explorer 4 2 Click the Next button to begin installation Click the Back button to change the installation settings 8 Click Next Page 8 of 50 LOLIGOSYSTEMS COM On Screen 6 the status of the installation is shown Screen 6 Saatos Ex Overall Progress Currently installing NI Assistant Framework LabVIEW Code Generator 6 1 Part 1 of 58 Po lt lt Back Next gt gt The installation may take a while When installation is complete Screen 7 becomes present Screen 7 Installation Complete The installer has finished updating your system lt lt Back Next gt gt Cancel 9 Click Next to end installation Page 9 of 50 LOLIGOSYSTEMS COM 10 If the USB hardware protection dongle drivers were not installed yet on the system then wait until you see Screen 8 Screen 8 hutoResp4 y You must restart your computerto complete this operation If you need to install hardware now shut down the computer if you choose to restart later restart your computer before
88. he path is by default the installation directory of AutoResp4 AutoResp4Data The current time and date xls is suggested as a filename in the prompt Both the path and filename can be altered by choice The system suggests two file extensions xls and txt The xls extension will create a tabulator separated text file which is readable by Microsoft Excel The txt extension will create a semicolon separated text file which is readable by any Microsoft Windows PC In addition to the data file the system will also generate a folder named as the data file The dot in the filename will however be replaced by an underscore _ in the folder name In this folder files containing raw data will be generated for each measurement loop The raw data files contain data from the measure period on which the calculated values in the main data file are based Please note The start experiment button will be disabled until a valid path filename is selected Page 33 of 50 L oii o 5 3 3 Setup field LOLIGOSYSTEMS COM 5 3 3 1 Flush period s Use this field to specify the duration of flush periods in seconds during the intermittent flow respirometry experiment Please note The field is limited to values between O s to 35 weeks 5 3 3 2 Wait period s Use this field to specify the duration of the wait periods in seconds during the intermittent flow respirometry experiment Please note The field is limited to values between 1 s to 35 weeks
89. he temperature in the ambient water as specified in the temperature control field 5 3 6 2 Setpoint C Use this field to set a temperature level in the ambient water that the system should maintain during an experiment Please note The field is limited to values between 2 C to 100 C 5 3 6 3 Hysteresis C Use this field to change the temperature hysteresis value Use a hysteresis value gt 0 C to ensure that the cooler heater pump does not flicker on the sharp edge of the setpoint If the hysteresis temperature 0 C the cooler heater pump is very susceptible to electronic noise on the temperature channel Please note The field is limited to values between 0 C to 49 99 C 5 3 6 4 Cool Heat Use these buttons to select if the system must maintain the setpoint as a maximum Cool or a minimum Heat temperature 5 3 7 Start experiment button Use this button to accept the setup settings as they are displayed on Screen 24 and proceed to Screen 25 5 3 8 Menus in Screen 21 Screen 23 and Screen 24 5 3 8 1 File gt Exit Exits AutoResp 4 without saving changes to the calibration screens 5 3 8 2 File gt Continue experiment Continues the last experiment if stopped by user Data on the graphs will only be in memory when AutoResp 4 has not been shut down Please note This option is only available if a calibration has been made and the user has stopped a running experiment 5 3 8 3 Help g
90. imited to values between 0 to 2500 I 5 3 4 2 Wet weight g Use this field to specify the weight of the experimental organism s Please note This field is limited to values between 0 001g to 100 kg 5 3 5 Oxygen control field 5 3 5 1 Checkbox Use the checkbox to enable or disable the O2 N relay on the DAQ 4 instrument If the checkbox is checked AutoResp 4 can control the O pressure in the ambient water as specified in the oxygen control field 5 3 5 2 Setpoint kPa Use this field to set an oxygen level in the ambient water that the system should maintain during an experiment Please note The field is limited to values between 0 kPa to 60 kPa 5 3 5 3 Hysteresis kPa Use this field to change the oxygen hysteresis value Use a hysteresis value gt 0 kPa to ensure that the O N valve does not flicker on the sharp edge of the setpoint If the hysteresis pressure is O kPa the O N valve is very susceptible to electronic noise on the ambient oxygen channel Please note The field is limited to values between 0 kPa to 30 kPa 5 3 5 4 Hypoxic Hyperoxic Use these buttons to select if the system must maintain the setpoint as a maximum Hypoxic or a minimum Hyperoxic oxygen pressure Page 35 of 50 L oii o LOLIGOSYSTEMS COM 5 3 6 Temperature control field 5 3 6 1 Checkbox Use the checkbox to enable or disable the TEMP relay on the DAQ 4 instrument If the checkbox is checked AutoResp 4 can control t
91. in chapter 7 6 3 3 Logging Measured data are logged during the measurement Add contents to the description fields in the File description section before starting logging max 3 lines This is not necessary but helpful as the description is later stored in the data file 1 Depending on the scan mode click on All channels for logging all channels or Single channel for single channel logging 2 Inthe appearing save as window specify filename without extension and its location The PC software automatically creates one to four different files The filename consists of the specified filename and the extensions ch1 txt ch2 txt to ch4 txt respectively ch stands for channel All filenames are displayed in the section Created files Please note Logging must be enabled before starting the measurement After the measurement it is not possible to store the measured values Starting logging during the measurement is also possible but stores only the values from the begin ning of logging on CREATED FILES SINGEL CHANNEL MAIN PATH ALL ACTIVE 1 2 3 4 CHANNELS f f ft 4 FILE Desenption ch 1 Description ch 3 DESCRIPTION 100 x a s 6 3 4 All Channels Graphical and numerical of all channel NOTE The All channels window is only enabled when scanning times for all adjusted channels are measured with the same scanning time The graphical display shows all data since the measurement was started The o
92. in the temperature cause variations in the measuring value at a constant oxygen con tent see Figure 12 11 Table 12 7 displays the deviation of the measuring value from the real oxygen content depending on temperature and the oxygen content Table 12 7 Deviations in the measured oxygen content A air saturation at a constant oxygen content and variations in temperature by 1K air saturation 25 air sat 100 air sat 250 air sat 50 9 hPa 203 6 hPa 508 9 hPa A air saturation K 0 43 0 88 hPa 1 62 3 30 hPa 3 91 7 96 hPa Example If the temperature is measured with a precision of 0 2 C there is a variation in the measuring value at 100 air saturation of 100 0 7 air saturation
93. is transmitted to the instrument via a polymer optical fiber The standard fiber cable length is 2 5 m If you need a cable length of more than 2 5 m up to 15 m available please con tact our service team Response times of the oxygen sensor OEC PSi3 Response time Stirred Not stirred Gaseous Oxygen too 60s 120s 20s Ordering information OIM Exchange Cap Order code for the OIM Exchange Cap OEC PSt3 Planar Oxygen Minisensor 5 2 6 OxyFinger Chemo Optical DO Probe for Mini Fermenters OFG PSt3 OxyFinger consists of a glass test tube glass finger which is coated with an oxygen sensitive foil The sensor signal is transmit ted to the instrument via a polymer optical fiber Cable lengths between 2 and 15 meters are available OxyFinger will be manufactured following your specifications Please specify both length and diameter of the glass finger and the size of the vessel closure Please contact our service team directly at info presens de The OxyFinger Chemo Optical DO Probe for Mini Fermenters has outstanding properties e The system can be used after autoclavation without recalibration e The system is fully autoclavable up to 100 times e In contrast to classical oxygen electrodes membrane cleaning and frequent replacement is not necessary e There are no electrolyte solutions to poison or replenish e No time for polarization is needed e Long shelf life e measuring range 0 250 96 a s e limit of
94. iven in the technical specification sheet Please read also the technical notes to avoid mistakes ELEMENT DESCRPTION FUNCTION ee ON OFF switch Switches the device ON and OFF C1 Line adapter for power Connector for 230 V AC power supply supply C2 RS 232 interface Connect the device with a RS 232 data cable to your male PC Notebook here Required Basic Equipment 5 4 Required Basic Equipment e Oxygen meter OXY 4 e Software for OXY 4 e PC Notebook System requirements Windows 98 2000 XP Millenium NT 4 0 Pentium processor at least 166 MHz 64 MB RAM a RS 232 port or a USB port and a USB serial adapter is needed e RS 232 Cable e Line adapter 110 220 V AC 12 V DC e Oxygen sensitive minisensor The minisensors are mounted into different types of housings e Vessels for calibration standard 100 water vapor saturated air 100 96 air saturation and calibration solution 0 oxy gen free water We recommend Schott laboratory bottles with a thread which can be obtained by Merck Eurolab ordering number 215L1515 scope of supply Planar Oxygen Minisensor 6 5 Planar Oxygen Minisensors 5 1 Sensor Characteristic of the Oxygen Sensitive Minisensors The principle of the sensor operation is based on the quenching of luminescence caused by collision between molecular oxygen and luminescent dye molecules in the excited state Figure 5 1 shows a typical response curve of an oxygen sensitive sensor
95. lectrodes membrane cleaning and frequent replacement is not necessary e There are no electrolyte solutions to poison or replenish e No time for polarization is needed e Long shelf life e measuring range 0 250 96 a s e limit of detection 0 15 96 a s Please note The OIM is only ready for use together with the Optical Exchange Cap OEC see page 16 containing the oxygen sensor membrane Ordering information Oxygen Inline Steel Tube Measurements Diameter mm Length mm 12 120 12 140 12 215 12 325 12 425 Planar Oxygen Minisensor 16 5 2 5 OIM Exchange Cap OEC PSt3 Applications The OIM Exchange Cap OEC is the sensitive coating in a metal cap It is used to replace the old sensitive coating and has outstanding properties e The PSt3 coated OEC can be used after autoclavation without recalibration e The PSt3 coated OEC is autoclavable up to 100 times e In contrast to classical oxygen electrodes membrane cleaning and frequent replacement is not necessary e The PSt3 coated OEC can be easily exchanged e There are no electrolyte solutions to poison or replenish e Notime for polarization is needed e Long shelf life e Optical isolation prevents cross sensitivity of the sensor towards turbid or fluorescent samples e measuring range 0 250 a s e limit of detection 0 15 a s The OEC is coated with the oxygen sensitive material PSt3 It has to be integrated into the Oxygen Probe OIM The sensor signal
96. lf must look toward the sample The figure on page 11 shows how the highly enlarged sensor spot must be glued to the vessel 3 Remove the protective cap from the male fiber plugs of the delivered fiber cable and connect it to the SMA plugs of the OXY 4 and the holding device The safety nut must be carefully attached while turning slightly clockwise If the oxygen sensitive foil is used in shaking flasks we recommend the use of coasters for shaking flasks They are shown below During calibration and measurement the sensor spot has to be placed on top of the fiber optic of the coaster The fiber optic of the coaster is located in the middle of the green positioning ring See also page 18 left Set of four coasters for shaking flasks right Coaster with green positioning ring Calibration 40 7 3 1 3 Performing the calibration 1 Switch on the OXY 4 instrument and the PC software following chapter 6 2 Goto window Calibration and enter the actual atmospheric pressure in hPa and the temperature of the calibra tion standards cal 0 and cal 100 The atmospheric pressure of the calibration is needed to convert the oxygen unit 96 air saturation into partial pressure units hPa Torr or concentration units mg L mol L Please ensure that there are no temperature changes during the calibration of the oxygen sensor Besides it must be ensured that the temperature during later measurement is constant and already known However the tempera tur
97. lso is based on equation 5 can be used for describing the oxygen calibration plot In this model Ksva is set to be m Ksy1 1 tanc f 1 f ee 8 tan IE Ks 0 Ix m K O The oxygen content in 96 air saturation can be calculated according to equation 9 B 4B 4 A C 2 A O with the coefficients Appendix 56 A tanc b ifi K2 9a tan tanc tan B in Kyy m Ks f m Ks 7 Ks f Ks 9b tan tan tano E tan D 9c 12 3 Oxygen Conversion Formulas Please note These conversion formulas are only valid in aqueous solutions and humidified air These formulas have to be modified if measurements have to be performed in organic solvents or solutions with high salinity saturation 26 air saturation Default setting of the instrument see equation 9 in 12 2 oxygen saturation 96 O air saturation ee 10 100 0 2095 volume content of oxygen in air ppm in the gaseous phase ppm O air saturation cil PR NIRE R07 100 10000 10000 11 ibn 1 Img E luL Z 1 1000000 1kg 1L 10000 Partial pressure of oxygen in hPa po hPa pan hPa py T hPa 2 25 mm 0 2095 12 in mbar Po mbar Pan mbar Pw T mbar ae D 0 2095 13 in Torr Po Torr i bati pot pabar 5n 0 2095 0 75 14 Please note 1 mbar 1 hPa 0 750 Torr Appendix Oxygen Concentration in mg L nicl rca
98. m OXY REG instrument When the button displays Fiber optic on the chamber oxygen input is acquired via the PreSens fiber optic instrument and the RS232 port of the PC Pressing the button toggles Screen 21 and Screen 23 seen on page 30 Before proceeding to Fiber Optic on the user is prompted to choose the COM port the fiber optic instruments from PreSens is connected too Please note When fiber optic is on as seen on Screen 23 on page 30 the sample frequency cannot be altered as it is predetermined by the fiber optic instrument from PreSens The frequency is approx 1 Hz when reading 1 channel and 2 Hz when reading all 4 channels 5 2 6 Set temp C Use this button to set the temperature of the ambient water in C This field is used only in fiber optic measurements and is disabled enabled together with the fiber optic button status OFF ON The default value is 20 C If the temperature cannot be kept during measurements within the entered value with 0 2 C choose to compensate the fiber optic oxygen signal with the measured temperature signal by enable the checkmark Analog Comp see 5 2 14 1 page 32 If Analog Comp is enabled the Set temp C field gets disabled and will be updated with the measured temperature The fiber optic oxygen value will now be real time compensated for the temperature Page 23 of 50 L oii o LOLIGOSYSTEMS COM 5 2 7 Next gt button Screen 22 Use this button to accept the calibratio
99. mixtures as well as in pure methanol and ethanol We recommend to avoid other organic solvents such as acetone chloroform or methylene chloride which may swell the sensor matrix Interferences were found for gaseous sulfur dioxide SO2 and gaseous chlorine Cl2 Both of them mimic higher oxygen concentrations Planar Oxygen Minisensor 8 Response time The response time too 90 96 of the signal change has occurred of the PSt3 oxygen sensor is less than 30 s in solution non stirred and even less than 8 s in the gas phase The response time too of the oxygen sensor is dependent from the diffusion rate of oxygen through the sensor layer and hence on the thickness of the sensor layer and the stirring rate A typical oxygen response curve of sensor membrane PSt3 in a non stirred and stirred sample solution is shown in Figure 5 3 below The response times too of sensor mem brane PSt3 are listed in Table 5 3 below Unlike electrodes optical sensors do not consume oxygen and the signal is independent of changes in flow velocity which means that stirring decreases the response time but has no effect on the measured value Optical isolation of the oxygen sensitive layer which is applied to exclude ambient light and improve chemical resistance will slow down the sensor response 100 oxygen minisensor PSt3 we 807 S 60 stirred too 40s B e non stirred a 40 T too 60s 3 J 20 0 0 20 40 60 80 100 120
100. n 1 and pin 4 GND in the input connector OXY AM Analog oxygen input for ambient water The input range is 0 5 VDC Connect analog oxygen input e g from an OXY REG via pin 1 and pin 4 GND in the input connector TEMP Analog temperature input for ambient water The input range is O 5 VDC Connect analog temperature input e g from a TMP REG via pin 1 and pin 4 GND in the input connector PC USB port for PC communication Use the USB cable to connect the DAQ 4 instrument to the PC LEDS Relay status LEDs IMPORTANT DO NOT connect DAQ 4 instrument to PC before installation of driver from Measurement amp Automation software from National Instruments is completed The software will be installed together with the installation of AutoResp 4 see page 6 Page 4 of 50 L oii o LOLIGOSYSTEMS COM 3 2 2 Back side FLUSH Relay socket for 110 230VAC max 3A flush pumps Connect one or more pumps by using one of the 4 fold power strips RECIRC Relay socket for 110 230VAC max 3A recirculation pump Connect one or more pumps by using one of the 4 fold power strips O2 N2 Relay socket for 110 230VAC max 3A for solenoid valve Use oxygen or nitrogen for hyperoxic or hypoxic control Connect e g a solenoid valve by using one of the accompanying device connectors TEMP Relay socket for 110 230VAC max 3A heating cooling pump Connect e g a heating cooling pump by using one of the acc
101. n absence and presence of oxygen lo and are the respective luminescence intensities O2 the oxygen concentration and Ksy the overall quenching constant bL zt eKsy 0 T f O 1 t f O5 I Luminescence intensity in presence of oxygen lo Luminescence intensity in absence of oxygen t Luminescence decay time in presence of oxygen To Luminescence decay time in absence of oxygen Ksv Stern Volmer constant quantifies the quenching efficiency and therefore the sensitivity of the sensor O2 Oxygen content Appendix 52 2 2 10 1 1 0 20 40 60 80 100 oxygen content Figure 12 2 A Luminescence decrease in the presence of oxygen B Stern Volmer plot Indicator dyes quenched by oxygen are for example polycyclic aromatic hydrocarbons transition metal com plexes of Ru Il Os II and Rh Il and phosphorescent porphyrins containing Pt II or Pd Il as the central atom 12 1 2 Major Components of Fiber Optic Minisensors In optical chemical sensors the analyte interacts with an indicator and changes its optical properties The result is either a change in the color absorbance or spectral distribution or the luminescence properties intensity lifetime polarization Light acts as the carrier of the information The major components of a typical fiber optical sensing system are e alight source to illuminate the sensor laser light emitting diode lamps e an optical fiber as signal transducer pla
102. n settings and move on to setting up a resp experiment see 5 2 14 page 32 Before proceeding to the setup the user is prompted to enter a data file name and path see Screen 22 The path specifies the location of the main data file and a folder containing raw data files from the experiment The path is by default the installation directory of AutoResp4MAutoResp4Data The current time and date xls is suggested as a filename in the prompt Both the path and filename can be altered by choice The system suggests two file extensions xls and txt The xls extension will create a tabulator separated text file which is readable by Microsoft Excel The txt extension will create a semicolon separated text file which is readable by any Microsoft Windows PC In addition to the data file the system will also generate a folder named as the data file The dot in the filename will however be replaced by an underscore _ in the folder name In this folder files containing raw data will be generated for each measurement loop The raw data files contain data from the measure period on which the calculated values in the main data file are based Save data file as Gemi 3 AutcResp4Data N 4 Seneste dokumenter Skrivebord Dokumenter Denne computer Filnavn 06 06 2007 1135 52 Netv rkssteder Fillype Excel compatible or delimited text file sls tx Page 24 of 50 L oii o LOLIGOSYSTEMS COM 5 2 8 Chambe
103. nn nnn natnm ssi nn snnt sns nn natnm sse tn sat nn nne nn annt 6 5 1 Sensor Characteristic of the Oxygen Sensitive Minisensors eese 6 5 2 Housings of Oxygen Sensitive Minisensors eeeeeeeeeeeeeeeeeeneenne nennen nennen 10 5 2 1 Planar Oxygen Sensitive Foils SP PSt3 ssssssssssseeeenenneenee nennen 11 5 2 Flow Through Cell with Integrated Planar Oxygen Sensor FTC PSt3 sssssesess 13 5 2 8 Oxygen Dipping Probe DP PSI3 sessesssssseseeeeeeeeneeen nennen nnne nnne nnne nnns 14 5 2 4 Oxygen Probe for Inline Measurements in Fermenters OIM 15 5 2 5 OIM Exchange Cap OEG PSIOQ ete te ete erties Haden tere rn cer EE eae 16 5 2 6 OxyFinger Chemo Optical DO Probe for Mini Fermenters OFG PSt38 cccssseeeeseteeeeeeeeees 17 5 2 7 Coaster for Shaking Flasks and Spinner Flasks CSF esee 18 6 SOMWAMG e 19 6 1 Software Installatiorn 2 e ree cde eren edes na Dee nee fenior i io esie ende crei de gue 19 6 2 Starting OXY 4 and its Softwaroe 5 eerie irure tectae internis ettari randa eanteeentetensteved 20 6 3 Function and Description of the OXY 4 Software eese nnne nnn 20 6 3 1 Me amp asSUretmmernt z s retten arate teet vtt eee epo quit e neben d im iribe Du EU ae 22 6 3 2 Galibrationoz zs ien oho EPA BR IE REB 22 6 3 3 boggin
104. nsor are known the Calibration with constant values is possible 1 Switch on the OXY 4 and the PC software following chapter 6 2 Go to the window Calibration Select the channel to be calibrated in the section Channel of sub window Single chan nel or switch to the sub window All channels 3 Enter the respective calibration values a phase at 0 air saturation temperature at 0 air saturation phase at 100 air saturation temperature at 100 95 air saturation pressure at which values a d were measured The atmospheric pressure of the calibration is needed to convert the oxygen unit 96 air saturation into par tial pressure units hPa Torr or concentrations units mg L mol L oco0c User defined calibration 55 00 327 00 4 To store the calibration values click the calibrate button A dialog appears and informs you that you will program all channels in case the sub window All channels is chosen If sub window Single channel is chosen a dialogue appears informing you that you will change the last calibration values Click the Continue button to store the new calibration data WARNING This will change last calibration values Cancel Repeat this procedure for all other channels by typing in the respective channel into the Channel Window to record user defined calibration values Calibration 38 7 3 Calibration of Planar Oxygen Sensitive Foils PSt3 Integrated in Glass Vials There are two ways to cali
105. ompanying device connectors 100 240VAC 50 60 Hz Power input for the DAQ 4 instrument Connect the instrument to a grounded 110 230VAC power supply using a standard pc type cable with a grounded wall plug Power button Turns the instrument on and off IMPORTANT DO NOT connect any of the relay sockets to gt 3 amps equipment USE grounded outlets only For further information about the DAQ 4 instrument see 8 3 DAQ 4 Instruction manual page 47 Page 5 of 50 LOLIGOSYSTEMS COM 3 3 AutoResp 4 software for Windows The following steps will explain how to install AutoResp 4 and drivers on your computer 1 Insert the CD labelled AutoResp 4 and wait until you see Screen 1 If you do NOT se the screen browse to the root of your CD and double click on the icon labelled setup exe Screen 1 Welcome to the AutoRespl 4 installer Please follow the instructions on screen and wait until you are asked to restart your computer ETE 2 Click Next Page 6 of 50 Screen 2 On Screen 2 the product information of the products to be installed are shown Product Information Please read the following information about the products to be installed NI DAQ 8 5 Device Support NI DAGmx 8 5 supports the latest DAQ devices including C Series M Series E Series S Series B Series USB Series Analog Qutput w aveform Counter Timer Digital 1 0 NI 653x DSA SCI and SCC devices The last version of NI
106. on 21 allows the solubility of oxygen in air saturated fresh water to be calculated for any temperature and pressure provided that the values of the Bunsen absorption coefficient o T and the vapor pressure pw T at the particular temperature are known Equation 22 or 23 can be used to obtain a and pw can be calculated from equation 20 The oxygen content cs of air saturated water can be calculated according to am Dy 8 Mo c pa 0 Pan Pw 5995 9 2 24 Pn Vy In equation 24 pam is the actual atmospheric pressure corrected for the contribution of the water vapor pressure Pw and related to standard pressure pw The corrected pressure is multiplied by 0 2095 the volume content of oxygen in air by a 0 and by the molecular mass of oxygen Moz divided by the molar volume Vy At a given atmospheric pressure of 1013 mbar Paim pn and a temperature of 20 C the oxygen content can be calculated according to equation 24 and results in 1013 23 3 32 g mol c LO13mbar 20 C 0 2095 0 031 0 009g L 9 06 mg L 25 1013 22 414mol L Table 12 3 gives oxygen solubilities in mg L for temperature intervals of 0 1 C from 0 40 C The calculated value for cs at a temperature of 20 0 C agrees with the tabulated value of 9 08 mg L Figure 12 9 shows the temperature dependent oxygen solubility in air saturated fresh water 15 14 13 12 11 10 Cg Oz mg L py pw T Py 0 2095 a 0 10 Mo Vy c
107. order to exclude ambient light from the fiber tip Schematic drawing POF polymer optical fiber L2 2 5 m sensor spot SMA connector 4 mm steel tube Features e usable for process application e very robust sensor with an excellent long term stability more than 100000 data points without drift e Ssterilizable H202 ethanol ethylene oxide e not autoclavable POF does not stand autoclaving conditions 130 C 1 5 atm e measuring range 0 250 a s e limit of detection 0 15 a s Ordering information Dipping Probe Oxygen Sensitive Coating PSt3 0 250 air saturation Order code for the Oxygen Dipping Probe DP PSt3 14 Planar Oxygen Minisensor 15 5 2 4 Oxygen Probe for Inline Measurements in Fermenters OIM OIM consists of a fitting made from stainless steel The oxygen sensor is integrated in the top of the metal fitting as shown below The metal fitting is connected to the instrument via a polymer optical fiber The standard fiber cable length is 2 5 m OIM is available in different sizes 12 mm 25 mm and standard OIM fits to B Braun Biostat B and B Braun Biostat C fer menters Polymer optical fiber Steel housing diameter 12 mm DO Probe immobilized on a glass support This chemo optical DO probe has outstanding properties e The system can be used after autoclavation without recalibration e The system is fully autoclavable up to 100 times e In contrast to classical oxygen e
108. oug D H2 AXO 9 1 J D umouq D ZH9 4X0 i Jerg umoug THO AXO A1ndNI HOSNSS es a D Jd i TA D L z ov A86 Se Ser D A ddns a Page 48 of 50 L oii o For using the LEDs and the relays see Table 4 and the wiring diagram on Figur 1 LOLIGOSYSTEMS COM Table 4 DAQ 4 connector NI USB 6215 pins Channel name line1 FLUSH 6 DO RECIRC 7 D1 O2 N2 8 D2 TEMP 9 D3 GND 5 n A Through the Solid state PCB in the DAQ 4 instrument it is possible to connect up to 500 W equipment to every relay SPECIFICATIONS Specifications range 20 C to 60 C Common specifications Supply voltage universal 21 6 253 VAC 50 60 Hz or 19 2 300 VDC Internal consumption Max consumption Isolation voltage test operation Signal noise ratio Response time programmable Calibration temperature Accuracy Temperature Coefficient EMC immunity influence Potentiometer input min Potentiometer input max Relay outputs Relay function Hysteresis in 9o display counts On and Off delay Sensor error detection Setpoint 0 3600 s Max voltage 250 VRMS Max current 2A AC Max AC power 500 VA Max current at 24 VDC 1A 3 2W 3 5 W 2 3 kVAC 250 VAC Min 60 dB 0 100 kHz 0 4 60 s 20 28 C lt 0 1 of reading lt 0 01 of reading C lt 0 5 of reading 10Q 100 kQ 0 1 25 1 2999 Make Break Hold Page 49 o
109. oxygen content in water outflow mg O liter BW body weight of experimental animal kg The advantages of this method are several 1 the duration of the experiment is in principle unlimited 2 no accumulation of CO and other metabolites 3 its possible to measure at a constant oxygen level 4 by controlling the quality of the inflowing water it s possible to measure metabolism at different desired levels of oxygen salinity etc However this method bring along one significant disadvantage in order to determine oxygen consumption by open respirometry it is crucial that the system is in steady state This means that the oxygen content of the in flowing and out flowing water AND the oxygen consumption of the animal have to be constant If the oxygen consumption of the animal for some reason changes during the experiment steady state will not exist for a while Not until the system is in steady state again will the above formula give the correct oxygen consumption rate The duration of the time lag depends on the relationship between chamber volume and flow rate Thus open respirometry measurements have poor time resolution and are not suitable for determination of oxygen consumption on organisms with a highly variable respiration like fish Page 17 of 50 L oii o LOLIGOSYSTEMS COM 4 3 Intermittent flow respirometry or open closed respirometry Our systems for automatic respirometry works by intermittent flow respirometry aiming
110. r oxygen field 1 4 5 2 8 1 Mov Avg button Use this button to select if the chamber oxygen input signal should be averaged or not The default value is 1 point which means no averaging of the input If any other value is chosen the chamber oxygen input will be averaged and the averaged values will be used for all subsequent computations screen and file outputs Use moving average if you wish to smoothen the chamber oxygen signal Please note No calibrated values will be displayed either on screen or in the data file before sufficient samples have been collected to compute the moving average E g if the moving average is set to 30 points and the sample rate is set to 10 Hz there will be no chamber oxygen values the first 30 samples equivalent to the first three seconds of samples 5 2 8 2 Input V indicator This field displays the actual voltage input 5 2 8 3 pO kPa indicator This field displays the actual calculated oxygen partial pressure from the voltage input 5 2 8 4 Lock LO button This button can be used to copy the actual value from the Input V indicator to the LO input V field instead of manually reading and retyping the value 5 2 8 5 LO input V Use this field to enter the low input voltage from the Input V indicator during the chamber oxygen calibration Please note The field is limited to values between OV to 5V 5 2 8 6 LO pO kPa Use this field to enter the oxygen pressure during the low l
111. rd The wizard has finished installing the software for 15 Click Finish Page 11 of 50 Screen 12 LOLIGOSYSTEMS COM Found New Hardware Wizard Welcome to the Found New Hardware Wizard Windows will search for current and updated software by looking on your computer on the hardware installation CD or on the Windows Update Web site with your permission Can Windows connect to Windows Update to search for software O Yes this time only Yes now and every time connect a device No not this time Click Next to continue 16 Click on No not this time and then on Next Screen 13 Found New Hardware Wizard This wizard helps you install software for USB 6215 If your hardware came with an installation CD lt 4 or floppy disk insert it now What do you want the wizard to do Install the software automatically Recommended Q Install from a list or specific location Advanced Click Next to continue Em NT 17 Click on Install the software automatically Recommended and then on Next Page 12 of 50 Screen 14 LOLIGOSYSTEMS COM Found New Hardware Wizard Completing the Found New Hardware Wizard The wizard has finished installing the software for 7 usBens Click Finish to close the wizard 18 Click Finish Screen 15 New Office Document E Newshorteut2 Qi Open Office Document LoiRe
112. ress this button to calibrate the OXY 4 fiber optic oxygen instrument in a 100 air saturation O sample Please note It is important not to push the Lock HI button before the Phase 9 and Amplitude indicators has stabilized Please note A full calibration must consist of a 0 calibration and a 100 calibration before the oxygen readouts are reliable Please note The calibration will be written down on an EEPROM in the OXY 4 instrument If you want to use the OXY 4 instrument with the software from PreSens a new calibration may be done using this software 5 2 13 7 HI Temp C Use this field to specify the temperature during calibration of the OXY 4 fiber optic instrument with the 100 air saturation O solution 5 2 13 8 Phase indicator This field indicates the phase angle of the optical signal that the OXY 4 fiber optic oxygen instrument uses for computations of the chamber oxygen pressure For further information about the OXY 4 fiber optic oxygen instrument see 8 4 OXY 4 Instruction manual page 50 5 2 13 9 Amplitude indicator This field indicates the amplitude of the optical signal that the OXY 4 fiber optic oxygen instrument measures The signal strength is an indicator for the sensor condition See the manual of the OXY 4 fiber optic instrument from PreSens for further explanation 5 2 14 Temperature field When the fiber optic button is ON the Temperature field gets changed from Screen 21 page 22 to Screen 23 p
113. river ff Startup E Vernier Software E winzip Kd Adobe Reader 7 0 Internet Explorer 0 0 v v v Y Y Y Y Y Y Y Y v v Y Y v Y Y Y Y Y LoliResp Microsoft Access Microsoft Excel Microsoft FrontPage B Microsoft Outlook Microsoft PowerPoint Microsoft Word Outlook Express po Remote Assistance e Windows Media Player a Windows Movie Maker j QuickTime A Windows Live Messenger gy wieu ev f LoiTRACK e Canon Utilities f AutoResp4 gram Files AutoResp4 Windows Home Edition e If the USB hardware protection dongle is not connected to the PC an error dialog box pops up see Screen 19 Screen 19 e Connect the USB hardware protection dongle to the PC via an USB port and click on eo Copy protection error No license found 2689 100000 Be nee Retry Screen 20 now becomes present Page 20 of 50 Screen 20 LOLIGOSYSTEMS COM Prec ven c M C e When AutoResp 4 has started properly click on the File menu gt New experiment Screen 21 is now present Page 21 of 50 5 2 Calibration and selection of oxygen input 5 2 Screen 21 AutoResp 4 File Edt view Help Sample frequency d 1 General about Screen 21 L oii o LOLIGOSYSTEMS COM Screen 21 is the
114. running any National Instruments software 11 Restart the computer but leave CD in the PC The following steps will explain how to install the DAQ 4 driver on your computer 12 Connect the DAQ 4 instrument to the PC via the USB cable After a few seconds Screen 9 becomes present Screen 9 s 3 Found New Hardware Wizard Welcome to the Found New Hardware Wizard Windows will search for current and updated software by looking on your computer on the hardware installation CD or on the Windows Update Web site with your permission Can Windows connect to Windows Update to search for software O Yes this time only Yes now and every time connect a device No not this time Click Next to continue 13 Click on No not this time and then on Next Page 10 of 50 Screen 10 Found New Hardware Wizard LOLIGOSYSTEMS COM This wizard helps you install software for USB 621x Loader lt 3 or floppy disk insert it now What do you want the wizard to do Click Next to continue C If your hardware came with an installation CD Install the software automatically Recommended Q Install from a list or specific location Advanced TT 14 Click on Install the software automatically Recommended and then on Next Screen 11 _ Found New Hardware Wizard Completing the Found New Hardware Wizard Q7 USBG2bx Loader gt Click Finish to close the wiza
115. s and converts the analog signals to 16 bit values Connect the inputs to controller instruments e g OXY AM to an OXY REG with a data cable If you want to use controller instruments not from Loligo Systems make sure the input range is 0 5 VDC The signal must be on pin 1 and GND on pin 4 on an input connector USING THE DAQ 4 IN OTHER APPLICATIONS It is possible to use the DAQ 4 instrument with other applications than AutoResp 4 See Table 3 for input connections and the wiring diagram on Figur 1 Table 3 DAQ 4 connector NI USB 6215 pins Channel name OXY CH 1 17 Ail OXY CH 2 19 Ai2 OXY CH 3 21 Ai3 OXY CH 4 24 Ai4 OXY AM 26 Ai5 TEMP 29 Ai6 GND 18 20 22 25 27 28 and 30 n A Page 47 of 50 5 d 2 D e z z dWaL 2N 20 3BI934 HSM aal m ATA a x i z i j ew 20 99 1 3 a ETA i i wo T lt j oromu m e a pera i A m x 3 z anraq HSM u MorpreA useJ6 i f ELA HINGANO TO LNOS i L Figur 1 LINE OUT nura OUTS Qur4 erqeo uoqqrH TM Solid state PCS YST E asna Y3 K1 Umoq m Lj Controller NI US8 6215 24 se ee oe at ISTRUTWJOL gt r eaj ajea USB A R P e o 4 9 ah 2 umaa 21 dial 5 JTA D uMouq D WY AXO eta Us UuModq gt YHO AXD m T J Dr uM
116. s and safety requirements The perfect functioning and operational safety of the instrument can only be ensured if the user observes the usual safety precautions as well as the specific safety guidelines stated in these operating guidelines Before connecting the device to the electrical supply network please ensure that the operating voltage stated on the power supply corresponds to the mains voltage The perfect functioning and operational safety of the instrument can only be maintained under the climatic conditions specified in Chapter 10 Technical Data in this operating manual If the instrument is moved from cold to warm surroundings condensate may form and interfere with the functioning of the instrument In this event wait until the temperature of the instrument reaches room temperature before putting the instrument back into operation Balancing maintenance and repair work should exclusively be carried out by a suitable qualified technician trained by us Especially in the case of any damage to current carrying parts such as the power supply cable or the power supply itself the device must be taken out of operation and protected against being put back into operation If there is any reason to assume that the instrument can no longer be employed without a risk it must be set aside and appropriately marked to prevent further use The safety of the user may be endangered e g if the instrument e is visibly damaged
117. s limited to values between OV to 5V 5 2 9 9 HI pO kPa Use this field to enter the oxygen pressure during the high level oxygen calibration of the ambient oxygen input Please note The field is limited to values between 0 00 kPa to 60 00 kPa 5 2 10 Temperature field 5 2 10 1 Mov Avg button Use this button to select if the temperature input signal should be averaged or not The default value is 1 point which means no averaging of the input If any other value is chosen the temperature input will be averaged and the averaged values will be used for all subsequent computations screen and file outputs Use moving average if you wish to smoothen the temperature signal Please note No calibrated values will be displayed either on screen or in the data file before sufficient samples have been collected to compute the moving average F g if the moving average is set to 30 points and the sample rate is set to 10 Hz there will be temperature values the first 30 samples equivalent to the first three seconds of samples 5 2 10 2 Input V indicator This field displays the actual voltage input 5 2 10 3 Temp C indicator This field displays the actual calculated temperature from the voltage input Page 27 of 50 L oii o 5 2 10 4 Lock LO button LOLIGOSYSTEMS COM This button can be used to copy the actual value from the Input V indicator to the LO input V field instead of manually reading and retyping the value 5 2
118. smaller than 0 05 and click the CAL 0 button to store the 0 calibration value at the adjusted temperature A message window opens and informs you that you will change the last calibration values Click the OK button to store the new calibration data Repeat this procedure for all other channels by typing in the respective channel into the Channel Window to record the oxygen free calibration value cal 0 6 Rinse the sensor membranes gently with water after calibration to remove sulfite remaining on the surface Calibration 32 7 1 2 User defined calibration If temperature and phase angle of a former calibration of the same sensor are known the Calibration with constant values is possible 1 Switch on the OXY 4 and the PC software following chapter 6 2 Go to the window Calibration Select the channel to be calibrated in the section Channel of sub window Single channel or switch to the sub window All channels 3 Enter the respective calibration values a phase at 0 air saturation temperature at 0 air saturation phase at 100 air saturation temperature at 100 air saturation pressure at which values a d were measured The atmospheric pressure of the calibration is needed to convert the oxygen unit 96 air saturation into par tial pressure units hPa Torr or concentrations units mg L pmol L oap 0 34 100 00 RH SNL NBN STI 56 00 Ib qmm 4 To store the calibration values click the
119. sp Microsoft Access Microsoft Excel SetProgram Access and Defaults E Microsoft FrontPage Wb Windows Catalog E Microsoft Outlook Ab Windows Update B Microsoft PowerPoint D winzip Microsoft Word Ei Accessories Outlook Express Ef Acer ePm Ei Acer Gridvista P E AcerSystem gb Remote Assistance Windows Media Player Windows Movie Maker E Antivir PersonalEdition Classic Uode Windows Liv C950 Multi Channel Gas Exchange Software A Windows Live Messenger Ei CyberLink PowerDVD wer Ef LoiTRACK E Eltima Software Ei Games Ef LABTECH NOTEBOOKpro Version 12 1 amp Quattro Track E Launch Manager Ei Measurement Computing amp LoliTRACK Ef Microsoft Office Tools fii National Instruments M f cel x en Es NT Backup Now 4 ES MORE Ef NTI CD amp DVD Maker 7 ui e Philips SPC SOONC PC Camera e amp Measurement amp Autd 2 Philips ToUcam Camera E Canon utilities E AutoResp4 a E mail ESE d Windows Home Edition gy Nag X Measurement amp Automation dy NI Registration Wizard GB Datasocket visa GA MI License Manager Ef Pinnade PCTv usB2 Paint REALTEK Gigabit and Fast Ethernet NIC Driver fg Startup Ef Vernier Software E winzip D Adobe Reader 7 0 4B Internet Explorer v v v v o 0 AR om o0 0 07 7 0 0 9 9 QxmguxBEume Sy sem 19 Open Me
120. stic or glass fiber e a photodetector photodiode photomultiplier tube CCD array e the optical sensor indicator immobilized in a solid matrix POF polymer optical fiber sensor spot L 2 5 m p SMA connector I amm D steel tube Figure 12 3 Scheme of a minisensor Appendix 53 Glasfaser Koppler Minisensor ST LED Figure 12 4 Schematic drawing of the optical setup of a measuring system with minisensors LED light emitting diodes PMT photomultiplier OF optical filters ST fiber connector 12 1 3 Advantages of Optical Oxygen Sensitive Minisensors e no oxygen is consumed during the measurement e the signal is independent of changes in flow velocity e they are able to measure the oxygen content in dry gases e they are insensible towards electrical interferences and magnetic fields e they are more sensitive than conventional electrodes up to ppt range e long term stability and low drift e using silica fibers it is possible to measure in samples while physically separate from the light source and detectors e light conducting fibers are able to transport more information than power currents information can be simul taneously transferred e g intensity of light spectral distribution polarization information such as decay time or delayed fluorescence 12 1 4 Luminescence Decay Time The OXY 4 measures the luminescence decay time of the immobilized luminophore as the oxygen dependent
121. t About Displays contact information about Loligo Systems Page 36 of 50 L oii o LOLIGOSYSTEMS COM 5 4 Running experiment 5 4 1 General about Screen 24 On Screen 25 the intermittent respirometry experiment is running Screen 25 E AutoResp 4 File Edit View Help Oxygen kPa SML uu quc uu m uw o uw m pq Loop remaining s 538 Status Fo Flush Recirc Go Chamber 1 O2 kPa 20 40 Ambient O2 kPa 120 65 Temperature C 197 99 Oxygen Temp Chamber 2 0 62 120 43 2 5 O n SR 14 40 36 14 50 36 us Chamber 3 Chamber 4 O2 kPa O2 kPa 80 2047 283 70 90 60 50 40 Current Temp C Chamber 1 ANANI 30 Chamber 2 MAN 20 Chamber 3 mE 10 Chamber ZZ Ambient v Cent 14 50 35 Temp nanan 14 40 35 5 4 2 Oxygen graph field This field shows the measured oxygen values vs time By right clicking the graph the user can choose which chamber graph is to be shown It is also possible to show the ambient oxygen as a graph and modify the time interval on the x scale 10 30 and 90 min Another option is to save the graph to a file and to show the experiment settings Page 37 of 50 L oii o LOLIGOSYSTEMS COM 5 4 3 Graph field This field shows some different graphs for the experiment As default the current temperature vs time graph is shown The user can choose between the following graphs by clicking with the mouse on the vie
122. t version at www presens de With best regards Your PreSens Team Appendix 51 12 Appendix 12 4 Basics in Optical Sensing of Oxygen 12 1 1 Dynamic Quenching of Luminescence The principle of measurement is based on the effect of dynamic luminescence quenching by molecular oxygen The following scheme explains the principle of dynamic luminescence quenching by oxygen ES Pd L emission of a light E 4 x s E absorption of light excited state energy transfer by collision no emission of light ES zm 4 a 4 Ao acm Figure 12 1 Principle of dynamic quenching of luminescence by molecular oxygen 1 Luminescence process in absence of oxygen 2 Deactivation of the luminescent indicator molecule by molecular oxygen The collision between the luminophore in its excited state and the quencher oxygen results in radiationless deactivation and is called collisional or dynamic quenching After collision energy transfer takes place from the excited indicator molecule to oxygen which consequently is transferred from its ground state triplet state to its excited singlet state As a result the indicator molecule does not emit luminescence and the measurable lumi nescence signal decreases A relation exists between the oxygen concentration in the sample and the luminescence intensity as well as the luminescence lifetime which is described in the Stern Volmer equation 1 Here to and are the luminescence decay times i
123. table measuring values are obtained 9 2 Maintenance The instrument is maintenance free The housing should be cleaned only with a moist cloth Avoid any moisture entering the housing Never use benzine acetone alcohol or other organic solvents The SMA fiber connector of the minisensor can be cleaned only with lint free cloth The sensor tip may be rinsed only with distilled water 9 3 Service Balancing maintenance and repair work may only be carried out by the manufacturer PreSens Precision Sensing GmbH Josef Engert StraBe 9 D 93053 Regensburg Germany Phone 49 0 941 942720 49 0 177 210 15 93 Fax 49 941 9427227 E mail info presens de Internet www presens de Please contact our service team should you have any questions We look forward to helping you and are open for any questions and criticism 10 Technical Data 10 1 General Data MODES PSt3 oxygen sensor CALIBRATION PROCEDURE PSt3 oxygen sensor OPTICAL INTERFACE Optical Connector Technical Data range 0 250 air saturation resolution 1 0 05 air saturation 30 0 1 air saturation 100 0 5 96 air saturation 250 1 7 96 air saturation accuracy 1 96 air saturation at 100 96 air saturation 2 point calibration in oxygen free water and humidified air or air saturated water SMA compatible 2 mm polymer optical fiber Wavelength 505 nm POWER INPUT 230VAC 50 60Hz 0 5A max DIGITAL INTERFA
124. ubbles located in the glass tubes of the flow through cells Wait about 3 minutes until the phase angles are constant the variation of the phase angles should be smaller than 0 05 and click the CAL 0 button to store the 0 air sat calibration values at the adjusted temperature After ward press the calibration solutions into the waste A message window opens and informs you that you will program all channels with the actual calibration values Click the OK button to store the new calibration data Rinse the sensor membranes gently with water after calibration to remove sulfite remaining on the surface Calibration 36 Calibrate Single Channels 1 Connect one plastic tubing with a syringe the other dip into the vessel containing the calibration solution 100 cal 100 Fill the syringe slowly with calibration solution cal 100 Please ensure that there are no air bubbles located in the glass tube of the flow through cell Measurement Calibration Logging All channels pE E Calibration constants 1100 00 41 20 0 J 20 0 Calibration with current values 99 80 1013 CAL 0 27 19 20 0 44265 200 CAL 100 0096 i ili 2 Wait about 3 minutes until the phase angle is constant the variation of the phase angle should be smaller than 0 05 and press the CAL 100 00 button to store the 100 air sat calibration value at the adjusted temperature Afterwards press the calibration solution into the waste
125. ust be ensured that the temperature during later measurement is constant and already known However the temperatures during the measurement and the calibration process are allowed to be different 3 Select the channel to be calibrated in the section Channel of sub window SINGLE CHANNEL or switch to the sub win dow ALL CHANNNELS Calibrate All Channels 1 For each sensor connect one plastic tubing with a syringe the other dip into the vessel containing the calibration solu tion 100 cal 100 Fill the syringes slowly with calibration standard cal 100 Please ensure that there are no air bubbles located in the glass tubes of the flow through cells Calibration SINGLE CHANHEL ALL CHANNELS 5 Ma air 2nd cal point 100 00 pressure standard Wait about 3 minutes until the phase angles are constant the variation of the phase angles should be smaller than 0 05 and press the CAL100 button to store the 100 air sat calibration values at the adjusted temperature Af terwards press the calibration solutions into the waste A message window opens and informs you that you will program all channels with the actual calibration values Click the OK button to store the new calibration data To record the second calibration values oxygen free water dip the plastic tubings into the vessel containing the cali bration solution 0 cal 0 and fill the syringes slowly with it Please ensure that there are no air b
126. ven for four different partial pres sures at 20 C the accuracy for two different partial pressures The resolution in oxygen is also transformed in different oxygen units Table 5 2 Oxygen resolution and accuracy of the PSt3 oxygen sensor membrane at different oxygen contents at 20 and 1013 mbar Resolution 1 0 05 96 air sat at 20 C and 1013 hPa 30 0 1 air sat 0 09 0 005 mg L ppm 100 05 air sat 2 72 0 01 mg L ppm 250 1 7 air sat 9 06 0 05 mg L ppm 021 0 01 oxygen 20 9 0 1 oxygen 52 4 0 35 oxygen 22 65 0 15 mg L ppm 63 002 oxygen 1 55 t 0 08 Torr 46 7 t 0 2 Torr 2 83 0 14 umol 155 5 0 75 Torr 85 0 0 28 umol 388 8 2 6 Torr 283 1 1 4 umol 2 0 1 hPa 798 0 4 7 umol 60 0 3 hPa 200 1 hPa 500 0 3 hPa Accuracy 20 C 1 at 100 air saturation 0 15 at 1 air saturation Temperature PreSens oxygen sensors can be used in the temperature range of 0 to 50 C The OXY 4 does not contain temperature sensors Temperature changes during the measurement are not compensated by the software Cross sensitivity No cross sensitivity exists for carbon dioxide CO2 hydrogen sulfide H2S ammonia NH3 pH any ionic species like sulfide So sulfate SO chloride Cl or salinity Turbidity and changes in the stirring rate have no influence on the measurement The sensors can also be used in methanol and ethanol water
127. w button in the menu MO vs time MO vs avg Chamber O5 Avg temperature vs time Current temperature vs time R vs time Current R vs time The MO avg temperature and the R vs time graph shows data from the whole experiment The current temperature and the current R vs time graph shows data for the last 10 30 or 90 min depended on the x scale from the oxygen graph see 5 4 2 By right clicking on the graph the user can save the shown graph to a file 5 4 4 Status field 5 4 4 1 Loop remaining s This field shows the remaining seconds left of the current measurement loop The loop is defined as the summed duration of flush wait and measure periods 5 4 4 2 Loop number s This field shows the actual state of the loop and the number of loops The letter F means flush W means wait and M means measure Behind the letter a number is shown which shows the actual number of the loop 5 4 4 3 Flush indicator This diode indicates if the flush pump is activated or not The flush pump is running only in a flush period 5 4 4 4 Recirc indicator This diode indicates if the recirculation pump is activated or not The recirculation pump is running in a flush and a measure period Page 38 of 50 L oii o LOLIGOSYSTEMS COM 5 4 5 Ambient measure field 5 4 5 1 Ambient O kPa This field shows the actual measured ambient water oxygen pressure 5 4 5 2 Temperature C This field shows the actual
128. wn for the chamber oxygen graph The state of the Play button also changes the menu of the Chamber oxygen graph see 5 2 11 page 29 Page 29 of 50 5 2 12 the PC General about Screen 23 Screen 23 is the calibration screen as it looks if the chamber oxygen signal is acquired from the OXY 4 fiber optic oxygen instrument and the RS232 port of L oii 0 LOLIGOSYSTEMS COM Please note When fiber optic is on as seen on Screen 23 the sample frequency cannot be altered as it is predetermined to approx 1 2 Hz by the OXY 4 instrument Please note The look and functionality of the Ambient Oxygen field and the Oxygen graph field are similar to those of Screen 21 Screen 23 P AutoResp 4 Help Sample frequency 10 Hz Chamber oxygen 1 Moy Avg i point YV LO Temp C Lock LO Jj 20 00 HI Temp C Lock HI J 20 00 HEE Jio Enable pO kPa 0 00 Phase 0 Amplitude 0 Barometic pressure hPa Chamber oxygen 2 Moy Avg 1 point x Lock LO Lock HI pO sat kPa yl 20 859 Enable pO kPa 0 00 LO Temp C Phase 1320 00 0 y HI Temp C Amplitude J 20 00 0 Chamber oxygen 3 Enable Mov Avg i point x LO Temp C LockLO 7 120 00 HI Temp C Lock HI 20 00 Jed Ambient oxygen Mov Avg Input V lpoint V 4 92 LO input v Jj 0 00 HI input V 5 00 y A Lock HI pO kPa 0 00
129. xygen unit is the same as specified in the window Measurement at the beginning of the measurement It is not possible to change the unit during the measurement With the data choice button it is possible to switch between the data a oxygen concentration in the unit specified in the window Measurement at the beginning of the measure ment b phase see appendix for description of phase and amplitude c amplitude see appendix for description of phase and amplitude The Clear graph button is used to clear the graphical window from all former data The scale of the y axis and the starting point of the x axis can be set to different modes by clicking on the Autoscale button Two options are implemented 1 Autoscale On The software determines the minimum and maximum values to display all data 2 Autoscale Off The maximum and minimum value of the y axis can be defined by the user By clicking on the current value the mini mum and or maximum value can be changed After typing the new value one has to click in the graphical window again to realize the change For the x axis only the starting value can be changed Zoom Function 1 Press the left mouse button and drag from left to right to enlarge a certain area of the graphical window The graphical window displays the selected data points and is not actualized with new data 2 Press the left mouse button and drag from right to left to recover the original display or click the Unzoom

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