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TD-700 Laboratory Fluorometer Operating Manual

1. BLK XXX units lt ENT gt Setup amp Cal Data Stream lt Yes gt No Press lt ENT gt then lt D gt to see Data Rate sec lt 1 gt 2 5 10 30 60 D Sen XXX Raw XXX XX Press ENT to see Oper XXX Hrs Pwr Level 10096 H TURNER DESIGNS T7 1B 10 96 600 Use the lt gt button on the keypad to toggle between options The option in brackets is the Selected option TD 700 Laboratory Fluorometer Operating Manual 2 Calibration Direct conc Mode 1 Mode 3 Units 2 Cal Procedure Use this screen to choose between the Simple mode and the other 2 modes Use this screen to choose between Raw Fluor and direct conc modes Use this screen to choose units to be displayed on the home screen Page 17 2 Raw Fluor Direct conc Simple Mode Mode Mode Insert Typical Insert Typical Maxrange XXXX Insert Blank Sample ENT Sample ENT 1 9 Change and press lt ENT y y y SETTING Sens Set Sample XXX Max conc XXX X Press lt 0 gt when Send Factor XX 1 OK 9 Change New ENT value stable y y Sensitivity Set y Number of Stds Reading Blank Sens Factor XX Sample XXX 1 5 X lt ENT gt WAIT New lt ENT gt y y y S
2. 5 Checkthatthe correct lamp is installed for your primary application See Section IV for details 6 Checkto see that the sample adaptor is the correct one for your needs Referto Appendix 6 for a detailed explanation of TD 700 Sample Adaptors 7 Reinsertthe Filter Cylinder lining up either the A B C or D applica tion indicators on the cylinder with the silver alignment mark on the inside rim of the sample chamber Replace the sample adaptor making sure that the adaptor is properly seated in the filter cylinder and close the sample chamber lid 8 Pluginthe unit Be sure to use only the Turner Designs power supply provided with your instrument It is critical to use the power supply provided by Turner Designs to meet EMI requirements 9 Turn on the power switch rear of unit and allow the unit to warm up for the countdown period 600 seconds 10 minutes NOTE For optimal instrument stability leave the instrumentturned on Page 5 TD 700 Top View Filter Cylinder Assembly TD 700 Side View TD 700 Laboratory Fluorometer Operating Manual 6 C General Precautions Do notleave the instrument reading OVER for an extended period of time several minutes Instrumentinstability can result If you are using the quartz halogen lamp do not touch the bulb It may be hot Also oils from your hand will affect the lamp s transmission When using solvents or other chemicals
3. Direct Conc 1 Mode 2 Cal Procedure 1 Setup 2 Calibration Insert Typical Sample ENT SETTINGSens 4 Sens Factor XX Sensitivity Set Sens Factor XX Sample XXX ENT Read amp Subtract Blank 1 Yes 9 No Co 1 point calibration in which a standard and optional blank are run in order to set the optimal range and sensitivity of the instrument The user chooses from a range of 100 950 on a scale of 1000 the relative magnitude of the standard The user also has the option to subtract the blank signal from all the readings To choose the Multi Optional Raw Fluorescence Mode press lt ENT gt from the HOME Screen press 1 for Setup then 1 again for Mode Use lt lt gt gt to choose the Multi Optional Mode Press lt ESC gt to return to the previous screen then press 2 to choose the calibration procedure Use the lt lt gt gt key to choose Raw Fluor for the Raw Fluorescence calibration procedure Press lt ESC gt twice to return to the Setup Cal Screen To access the calibration sequence press 2 from the Setup Cal Screen The Multi Optional Raw Fluorescence calibration sequence will appear Fill a clean test tube or cuvette with a sample that you wish to read You do not need to know the exact concentration you are using it to set the optimal instrument sensitivity and range Wipe the outside of the test tube or cuvette dry and insert it into the sample adaptor in th
4. TD 700 Laboratory Fluorometer Operating Manual Dated 5 8 2002 Version 2 0 Part Number 7000 998 TURNER DESIGNS Kc laude Sunnyvale CA 94085 Less 09 E 740 004 PAX A 08 749 0998 imerdesigns com Toll free 1 877 316 8049 TD 700 Laboratory Fluorometer Operating Manual TD 700 Laboratory Fluorometer Operating Manual Dated 5 8 2002 Version 2 0 Part Number 7000 998 TURNER DESIGNS Kc laude Sunnyvale CA 94085 Less 09 E 740 004 PAX A 08 749 0998 imerdesigns com Toll free 1 877 316 8049 TD 700 Laboratory Fluorometer Operating Manual Page 2 VI VII VIII TD 700 Laboratory Fluorometer Operating Manual Table of Contents Introduction A Description B Inspection and Setup C GeneralPrecautions D Definition of Symbols Hardware Overview A TD 700 Quick View Diagram TD 700 Controls Optical Filter Installation and Removal Lamp Installation and Removal A TheMercury Vapor Lamp B TheQuartz Halogen Lamp Instrument Parameters Firmware Power up Screen HOME Screen Setup Calibration Screen Setup Screens Calibration Screens Firmware Flow Chart mnmmoous Calibration Overview A WhyCalibrate B WhentoCalibrate C TD 700 Calibration Options Calibration Simple Mode Calibration Multi Optional Mode Raw Fluorescence Calibration Multi Optional Mode Direct Concentration ToCalibrate How Sample Concentrations Will Be Calculat
5. Max Range XXXX 1 OK 9 Change Max conc XXX X New ENT Number of Stds 1 5 X lt ENT gt Enter highest conc Sid first Hi Std Conc XXX X 1 OK 9 Change L Continued on next page IX Calibration Multi Optional Mode Direct Concentration The Multi Optional Mode Direct Concentration calibration procedure is a multi point calibration in which one to five standards plus a blank are run in order to determine the concentration of an unknown This procedure will also set the optimal range and sensitivity of the instrument The Direct Concentration procedure should be used if you require superior accuracy and wish to determine the actual concentration of an unknown The TD 700 will display actual concentration in the units chosen during setup A Calibrate 1 To choose the Multi Optional Direct Concentration Mode press lt ENT gt from the HOME Screen press lt 1 gt for Setup then lt 1 gt again for Mode Use lt lt gt gt to choose the Multi Optional Mode Press lt ESC gt to return to the previous screen then press lt 2 gt to choose the calibration procedure Use the lt lt gt gt key to choose Direct Conc forthe Direct Concentration calibration procedure Press lt ESC gt to return to the previous screen then press lt 3 gt to choose the units of measure Use the lt lt gt gt key to choose the required unit of measure Press lt ESC gt twice to return to the Setup Ca
6. 1 Press D from the HOME screen when the data stream feature is on 2 Use the lt lt gt gt key to toggle between numbers TD 700 Laboratory Fluorometer Operating Manual Page 30 APPENDIX 1 Before Calibration or Reading Samples A Materials Needed 1ormore cuvettes 25 mm test tubes 13 mm test tubes or 10 mm squarecuvettes ABlank sample water or liquid containing none of the fluorescent material except when using the instrument in Simple Mode Standard s 1 to 5 solutions of varying but known concentrations for the Direct Concentration Calibration Procedure OR for the Simple mode one standard that is approximately half the level of the highest sample you are likely to read used to set the optimal sensitivity and range ofthe TD 700 Samples to be read B When Handling Samples 1 Takecare notto spill samples into the sample chamber Wipe up any spills promptly See Appendix 9 Section A for details on mainte nance 2 Rinse the test tube or cuvette 3 times between samples preferably with the sample to be read The TD 700 is very sensitive and even small amounts of material from a previous sample may result in errors Complete rinsing is especially important if you are using the same test tube or cuvette for samples and blank 3 Fill the test tube or cuvette at least 75 full significant error will result if it is not full enough 4 Thecuvette MUST BE DRY on the outside when taking r
7. To install use adapters 1 Open the lid to the sample chamber 2 Align pin or point arrow on the adapter to the silver dot on the TD 700 3 Slidethe sample adaptor inside the sample chamber and push down until it is fully seated in the chamber To remove sample adapter 1 Openthe lid to the sample chamber 2 Removethetesttube 3 Graspthe sample adaptor by the rim and pull straight up TD 700 Laboratory Fluorometer Operating Manual Page 41 Minicell Adapter The minicell cuvettes allows for very small volumes 75 2504 to be analyzed To install 1 Slide minicell adapter into the 10x10 mm cuvette adapter where a normal cuvette would sit 2 Insert the borosilicate glass minicell into the adapter 3 Insert into the TD 700 as described earlier TD 700 Laboratory Fluorometer Operating Manual Page 42 APPENDIX 7 Glossary Accuracy The degree to which a measured result approximates the true value ofthe quantity being measured This value is usually expressed as a percent Analyte The substance you wish to measure Attenuator A part used to cut down the amount of light that enters and exits the sample Can be used to adjust the sensitivity and dynamic range of your analysis Blank Sample water that contains none of the fluorescent material to be measured Blank is used during calibra tion to set the instrument to zero Calibration Setting the sensitivity of the instrument to the standard and z
8. atoms and molecules It is a molecule s ability to absorb light energy at one wavelength then instantaneously re emit light energy of another usually longer wavelength Each compound that fluoresces has a characteristic excitation wavelength the wavelength of light that it absorbs anda characteristic emission wavelength the wavelength of light that it emits when the molecules relax and return to their ground state These excitation and emission wavelengths or spectra are often referred to as the compound s fluorescence signature Figure 2 shows the key components of a filter fluorometer Digital Readout Light Detector Wavelengths specific to compound Emission Filter Wavelengths created by compound plus stray light Lamp Cuvette or Sample Cell N Excitation Filter Soecitic wavelengths of light Many wavelengths of light Figure 2 Principles of Filter Fluorometry Lamp The lamp or light source provides the light energy that excites the compound of interest The lamp actually provides a broader range of light than that which excites the compound This broad light range is illustrated by the many wavelengths of light shown in Figure 2 TD 700 Laboratory Fluorometer Operating Manual Page 52 Excitation Filter The excitation filter is used to screen out the wavelengths of light not specific to the compound you are measuring This filter allows a smaller band of light energy to pass th
9. higher valueforthe Maximum Range Note 1 If the sensitivity exceeds the maximum before reaching the target sensitiv ity a note message will appear The unit will automatically adjust sensitiv ity to the maximum If sensitivity falls below the minimum before reaching the target sensitivity the unit will automatically adjust sensitivity to the minimum Note that this may change the Max range concentration you entered Error3 The concentration entered for 42 5 Std is greater than the concentration entered for the highest Std Press ENT and enter a lower concentration for the Std OR press ESC then 1 to abort the calibration Begin again using the highest concentration standard first Error4 Fluorescence reading for 42 5 Std is higher than the reading for the highest standard Press ESC to abort the calibration rerun using the highest concentration standard first Error5 Fluorescence reading for blank is higher than the reading for one or more of the standards Press ENT to rerun blank make sure blank is OK and rerun OR press lt ESC gt to abort the calibration calibrate again using standards more concentrated than blank o 45 Invalid input lt ENT gt to reenter Error1 Std conc maxrange Note1 MAX or MIN sens Press ENT Error3 Std conc ent gt Hi Std Error4 X Std reads gt Hi Std Error5 Blk reads gt Std X TD 700 Laboratory Fluorometer Operatin
10. instrument is currently configured See figure to the left Simple Mode 1 Mode To access the Setup Screen from the Home screen press ENT then 1 Press the appropriate number to access the setup parameters Raw Fluorescence 1 Mode This parameter allows the user to choose between Simple amp 1 Mode Multi Optional calibration modes See Section V E for more details 2 Cal Procedure 2 Cal Procedure This parameter allows the user to choose between Raw Fluorescence and Direct Concentration calibration procedures See Section V E for more details Direct Concentration 3 Units Used only in Multi Optional Direct Concentration mode only This is to choose which units your standards and samples are 1 Mode 3 Units measuredin 2 Cal Procedure For all 3 of these parameters use the lt gt key to toggle between choices When you have your choice selected press ESC or H E Calibration Screens The instrument will initiate the calibration process when 2 of the keypad is pressed from the Setup Calibration Screen The firmware will guide the user through the process Depending upon the calibration mode and procedure chosen in the Setup screens the steps will be different See firmware flowchart on pages 16 amp 17 Here is a description of the 3 different calibration procedures Simple One point calibration This calibration allows you to use one standard or sample and no blanks The instrument g
11. or the lamp will not work 5 Replace the lamp access panel and push in the black quick release latches to snap the panel in place 6 Turn on the power Check the lamp view port to ensure the lamp is lit Also make sure the fan is operating If the fan is not operating check the connector plug TD 700 Laboratory Fluorometer Operating Manual Page 11 Quick release Lamp View Port TD 700 with Mercury Vapor Lamp Rear View Page 12 SE B The Quartz Halogen Lamp aiches 2 The average halogen lamp life is 2000 hours e o o lo EB e 1 e e J 2 LO TD 700 with Halogen Lamp Rear View 1 AN Check the lamp if The instrument is not responding even though the unit is plugged in and the power is on The readings are very low unstable or drifting To replace the quartz halogen lamp Turn off the instrument and remove the lamp access panel by pulling on the two black quick release latches on the rear panel of the instru ment Unplug the power jack that connects the lamp panel assembly to the instrument The lamp access panel should now pull free of the instru ment DO NOT TOUCH THE LAMP BULB with an ungloved hand the bulb may be hot and or the oils from your hand will affect the lamp s transmittance and cause incorrect readings If the bulb is burned
12. three terms are synonymous and are used to indicate the TD 700 fluorometer s sensitivity on a scale of 1 to 100 Sens Factor is the term used in the software during calibration Sens 96 can be found on the diagnostic screen and sensitivity level is the term used on the printout The Sensitivity Factor is directly proportionalto the voltage on the photomultiplier tube The higher the Sensitivity Factor the greater the sensitivity and typically the greater the noise of the readings The basic operating level of the instrument It is related to the detection limits the dynamic range and the resolution The electronic sensitivity is set during calibration using the keypad Sensitivity can also be adjusted mechanically using different cuvette sizes APPENDIX 8 Error Messages and Notes During calibration the TD 700 will display an error message if incorrect values are entered or if the readings may result in an inadequate calibration A Invalid Input This screen will appear when you enter a value that is outside the range for the parameter For example if you enter a number greater than 1000 for the Max range you will see Invalid Input Errori The concentration entered for the standard Std is greater than the maximum concentration entered for the Maximum Range Press ENT and enter a lower concentration for the Std OR press ESC then 1 to abort the calibration use a lower concentration standard or enter a
13. 227 9 Calibration Curve Data Conc Range Fluor Range Slope O BLK 1 500 0 00 6 710 0 223 1 500 6 200 6 710 29 09 0 210 6 200 50 00 29 00 2279 0 220 Slope RSD 3 127 An Explanation of the Printout The first four lines of the printout display the TD 700 s current parameters the mode the cal procedure the units and the instrument sensitivity level The sensitivity level a number between 1 and 100 is a measure of the photomultiplier voltage Next is the calibration that is stored in the unit s memory The maximum concentration range entered during the calibration sequence is displayed here Following this is the Calibration Standard Summary which displays each standard its concentration and its corre sponding raw fluorescence value Concentration and raw fluorescence values are organized in the Calibration Curve Data table so that the slope of the line between each data point is displayed The percent relative standard deviation RSD is displayed below this The RSD is an indicator of how well each of the slope values correlate to one another A consistent slope a slope that is almost the same for all data points will result a low RSD A nonlinear set of data will result in a high RSD The RSD can be thought of as an overall indicator of the data quality A RSD of less than 25 is considered by many to be acceptable while a RSD of less than 5 indicates excellent linearity Both the individual slopes and av
14. Laboratory Fluorometer Operating Manual 10 TD 700 Controls 1 10 11 Display Shows the screens and readout for samples It is lit continuously when the unit is operating Except during calibration the contrast of the display can be adjusted on any screen by pressing the up or down arrow keys Keypad The keypad is used to enter new values and to move through the software screens Sample Chamber Samples are placed inside the chamber for reading It houses the Filter Cylinder and the cuvette adaptor Filter Cylinder The Filter Cylinder has openings for various optical filter combinations It can be removed by grasping the inside rim of the cylinder and pulling it straight up Sample Adapters Adapters are available for various test tube or cuvette sizes The sizes used with the TD 700 are 25 mm round 13 mm round 10 mm square or 100ul minicell Lamp Access Panel To access the lamp remove the panel by pulling on the two black quick release latches on the lamp access panel The lamp provides excitation light at wavelengths specific to the compound to be measured Power Switch ON OFF switch RS 232 Serial Port Connection RS 232 DB9 female from the TD 700 serial port for connecting to a computer or printer Power Plug Socket The power supply connects into the rear panel ofthe instrument Drain Plug The TD 700 is equipped with a non removable plug located underneath the i
15. NT From the HOME screen press 9 to return to the Power up Screen HOME Screen After the countdown period the HOME screen is displayed The HOME screen is where samples are read and data is sent to the printer or computer Press H to return to this screen from any screen except during calibration From the HOME screen several functions can be accessed Press D to send data to a printer or computer Press to Discrete Sample Average See Section X for details Press 0 to Autozero or blank the instrument See Section X for details Press 8 for Diagnostic Screens Press 7 for Data Stream Setup Calibration Screen The Setup Calibration screen accessed from the HOME screen by pressing ENT is a screen where the user selects either to go to the calibration sequence or to review or change the setup parameters The user may also press D from this screen to send the current calibration information to a printer or computer TD 700 Laboratory Fluorometer Operating Manual Page 13 TURNER DESIGNS T7 1B 10 96 600 A Power up Screen XXX ppm ENT Setup amp Cal B HOME Screen 1 Setup 2 Calibration C Setup Calibration Screen Page 14 D Setup Screen This screen is used to choose the calibration procedure for the TD 700 Simple Multi Optional Raw Fluorescence Multi Optional Direct Concentration This screen will have 1 of 3 looks depending upon how the
16. acing the instrument place the solid standard in the filter cylinder with the letter L etched on top of standard holder on your left hand side Standards are marked with the letter L to designate low concentration and H to designate high concentration 4 Press lt gt to read value Record this number on the label located at the bottom of the standard as the low concentration L 5 Now pull standard out of filter cylinder rotate standard 180 degrees and place in filter cylinder again so that the letter is on your left hand side 6 Press lt gt to read value Record this number on the label located at the bottom of the standard as the high concentration 7 Removesolid standard and replace any cuvette adaptors that were removed in step 2 8 Record instrument settings on TD 700 Calibration Record with Solid Standard sheet that has been included for future reference 9 Proceed with analyzing your samples 10 You may use your solid standard at any time to check the stability of the instrument Simply remove any cuvette adaptors and insert your standard Press lt gt to read The value should be similar to what was previously obtained in steps 4 and 6 above 11 Also you may use it to calibrate on future days using the values obtained in steps 4 and 6 above For example if the values obtained for L and were 50 and 100 respectively you can use both to calibrate your instrument on
17. ample XXX y Enter highest Calibration lt ENT gt SETTING Sens conc Std first Complete Send Factor XX y y y Printing Cal y Hi Std Conc XXX X Printing Cal Report pls wait Sensitivity Set 1 9 Change Report pls wait Sens Factor XX Y Hi conc Sample XXX New lt ENT gt lt ENT gt Insert Hi Std Read amp Subtract and press Blank 1 Yes 9 No Y y Press 1 to see Insert Blank and press ENT Y Press ENT to see Press 0 when value stable Y Press 0 to see Reading Blank Wait y y Printing Cal Report pls wait SETTING Sens Send Factor XX y Sensitivity Set Sens Factor XX y Reading Hi Std WAIT y X Std Conc XXX X 1 OK 9 Change y X conc XXX X New lt ENT gt Insert X Std and press lt gt Y Reading X std WAIT TD 700 Laboratory Fluorometer Operating Manual Page 18 Note Calibration data will be stored even after power to the instrument is turned off Notethata separate calibration is stored for each of the three modes For example if you calibrate in the Simple Mode this calibration remains in effect until you recalibrate in the Simple Mode If you calibrate in the Multi Optional Mode Direct Concentration this will not change the cal
18. another day When calibrating enter in 100 for your first standard concentration and 50 for your second concentration L If calibrating with only one of the concentrations enter in the corresponding value 50 or 100 depending on whether L or H is on your left side when facing the instrument TD 700 Laboratory Fluorometer Operating Manual Page 54 Date Instr S N TD 700 Calibration Record with Solid Standard Calibration Std Conc s Units Mode Simple or Multi Cal Procedure Raw or Direct Maximum Range W Reading Reading TD 700 Laboratory Fluorometer Operating Manual
19. ations amp Features 49 APPENDIX 11 Principles of Fluorescence 51 APPENDIX 12 Instructions for Using the Solid Standard 53 TD 700 Laboratory Fluorometer Operating Manual Page 4 NOTES TD 700 Laboratory Fluorometer Operating Manual Introduction Description The TD 700 Fluorometer is a compact laboratory fluorometer designed for discrete sample measurement of various fluorescent materials including chlorophyll rhodamine fluorescein histamine vitamins and other fluores cent compounds When properly calibrated TD 700 will electronically set the optimal sensitivity and range for your samples and will readout the actual concentration of the compound when in the Direct Concentration mode The TD 700 can be set up to detect the specific compound you want to measure simply by changing the lamp and optical filters Inspection and Setup Upon receiving your fluorometer please inspect it carefully and make sure all accessories are present refer to the packing list shipped with the instrument To get started Refer to the Quick View Diagram page 7 1 Locate the instrument on a flat surface with the back of the instrument well ventilated at leastthree inches from a wall 2 Openthe sample chamber lid located on the top of the unit 3 RemovetheFilter Cylinder by grasping the inside rim of the cylinder and pulling up 4 Install the correct optical filters for your application See Section III for details
20. conds TD 700 Laboratory Fluorometer Operating Manual Page 43 Drift The change in measurement of the same sample over time Dynamic Range maximum range The range of concentrations that can be read by the TD 700 atthe current level of sensitivity Since zero is frequently assumed to be the minimum dynamic range in this manual often refers to the maximum concentra tion that can be read The user can select the desired maximum Max Range during calibration in the Direct Concentration calibration procedure Fluorescence The TD 700 Fluorometer measures the concentration of various analytes in samples of interest via fluorescence A fluorescent molecule has the ability to absorb light at one wavelength and almost instantly emit light at a new and longer wavelength A fluorescent molecule has a unique fluorescence signature which allows one substance to be distinguished from another In a fluorometer light from a lamp is passed through an excitation filter that transmits light of a wavelength range specific to the sample compound being measured The light passes through the sample which emits light proportional to the concentration of the fluorescent material present and proportional to the intensity of the exciting light The emitted light is passed through an emission filter that selects for the appropriate wave length range and the resultant light is then detected by aphotomultiplier tube Gain An indicator of the sensiti
21. d do not wish to recalibrate When resetting the blank to zero you are replacing the last blank setting with the new blank setting Again this procedure will not compensate for errors in sample readings due to instrument drift For maximum accuracy you should recalibrate 1 Inserta blank then press 0 from the HOME screen 2 Press 9 to abort the procedure or 1 to Reset Blank to Zero Blank Press 1 and without further prompting the instrument will blank to 1 Yes 9 No zero i 3 If the reading for the blank exceeds the maximum the unit will display BLK XXX ppm an error message Blank level too high If this message should ENT Setup amp Cal appear evaluate your sample to ensure that you are reading the blank and not another type of sample Once you have verified that you are reading the blank try the procedure again If the message persists recalibrate the instrument Data Stream This feature allows you to output data directly to a printer or a computer at regular intervals Only the value seen on the LCD screen will be printed No Data Stream index or raw fluorescence value will be printed Yes No 1 Press 7 from the HOME screen 2 Press c to toggle between Yes and No turns data stream on or off When the data stream feature is on the data rate can be changed The number selected is the time interval between data output Data Rate sec lt I gt 2 5 10 30 60
22. different measurements depending upon their orientation in the sample adaptor This is due to defects in the shape of the test tube that are not visible to the human eye We recom mend that the test tube be marked at the top and positioned in the adaptor the same way each time to minimize error F Data Quality The TD 700 is only as accurate as the standards that are used to calibrate it This is why it is important to take care when preparing standards samples and blanks One should follow good laboratory practices when preparing these standards TD 700 Laboratory Fluorometer Operating Manual Page 33 APPENDIX 2 The Calibration Printout A Printout Capability with a Printer or Computer To use this function with a printer 1 Connectthe TD 700 to a serial printer via the RS 232 port on the back ofthe instrument If you ordered a printer from Turner Designs the appropriate serial cable is included 2 Press D or from the HOME screen and the readout currently displayed will be sent to the printer To use this function with a computer 1 Connectthe TD 700 to your computer via the RS 232 port on the back of the instrument Use the DB9 serial cable included with your instru ment 2 The TD 700 signal is in ASCII format so you can use various communi cations programs to import data to your computer On your computer run the communications program you have chosen For a step by step procedure using Microsoft Windo
23. e the TD 700 will automatically adjust to the optimal range for sample measurement If the sample used is too concentrated or too dilute the instrument may not be able to reach the target sensitivity In these cases a note message will appear indicating that based on the calibration the unit has reached its sensitivity maximum or minimum By pressing lt ENT gt you tell the instrument to accept the maximum or minimum sensitivity value It is recommended however that you adjust your sample concentrations to fall within range and then recalibrate To view the last calibration data set press ENT from the HOME screen then 9 to view your blank and calibration standard value Press H to return to the HOME screen To print the last calibration data set press ENT from the HOME screen then D Press lt H gt to return to the HOME screen To abort the calibration press lt ESC gt at any time during the calibration sequence Press 1 to abort or lt ESC gt to resume TD 700 Laboratory Fluorometer Operating Manual Page 21 NOTES TD 700 Laboratory Fluorometer Operating Manual Page 22 Calibration Multi Optional VIII Raw Fluorescence 1 Setup Calibration Multi Optional Mode Raw Fluorescence 2 Calibration The Multi Optional Mode Raw Fluorescence calibration procedure is a single 1 Mode 2 Cal Procedure Simple lt Multi Optional gt 1 Mode 2 Cal Procedure lt Raw Fluor gt 2
24. e filters excitation and emission filters for the application you want to run Filters are marked on their rim with an identification number c The Filter Cylinder has eight openings for four different filter sets For example the openings for filter set A are marked A EX and A EM filter set B is marked B EX and Select the A B C or D positions to install the filters d Findthe openings for the filter set you want to remove For example if you want to install the filters in position B then remove any filters from the B EX and B EM openings To remove a filter take out the flexible rubber o ring holding the filter in place Use a tool such as a plastic pen cap or plastic nosed forceps to do this Be careful not to scratch the surface of the filter Place your hand over the opening and tilt the cylinder so the filter drops out into your hand Be careful the filters are glass and may break if dropped e Locate the excitation filter to be installed Handle the filter on the edges so as to avoid leaving fingerprints on the filter or wipe off the filter before installing If the filter has one side that is mirrored or highly reflective it should be installed so the mirrored side faces outward from the cylinder toward the lamp Insert it mirrored side out if any in the opening marked with the set letter EX Push the filter in so it rests flush with the back of the opening Then reinsert the o ring and press it
25. e sample chamber Press ENT to proceed to the next screen Default setting raw fluorescence reading is 800 Ifthis is acceptable press 1 to continue If you want to change this value press 9 Type in the new value between 100 950 and press ENT Press 1 to continue The TD 700 will now set its sensitivity as indicated by the SENS FACTOR sensitivity factor based on the final sample value that you accepted Once the sample is set the TD 700 asks whether you want to run a blank If you wish to have blank subtracted press 1 If not press 9 and the calibration sequence will end here and you will be returned to the HOME screen At this point the calibration data will printout automatically to a printer or a computer See Appendix 2 for details regarding the calibration printout TD 700 Laboratory Fluorometer Operating Manual 10 If you choose to run a blank fill a clean test tube or cuvette with the blank solution wipe the outside of the test tube or cuvette dry insert it into the sample adaptor in the sample chamber and press ENT Allow the reading to stabilize and press 0 The unit will read the blank then automatically return to the HOME screen At this point the calibration data will printout automatically to a printer or a computer See Appendix 2 for details regarding the calibration printout Errors After reading the sample the TD 700 will automatically adjust to the optimal range for
26. eadings Moisture on the outside will result in error C Linear Range and Quenching The linear range is the concentration range in which the readout of the TD 700 is directly proportional to the analyte concentration The linear range begins with the smallest detectable concentration and spans to an upper limit concentration that is dependent upon the properties of the fluores cent material the filters used and the path length Beyondthe linear range above the upper concentration limit the fluores cence readings rise at a decreasing rate and a calibration curve is neces sary for accurate readings At even higher concentrations readings will begin to decrease even though the sample concentration is increasing This effect is known as concentration quenching TD 700 Laboratory Fluorometer Operating Manual Page 31 For example Rhodamine WT fluorescent dye provides linear readings from about 10 parts per trillion to approximately 0 1 ppm Using the 25 x 150 mm Sample Adaptor linearity should be checked when measuring con centrations of dye higher than 0 1 ppm At dye concentrations below 0 1 ppm a single point calibration one standard and a blank may be used to calibrate the TD 700 For concentra tions between 0 1 and 0 5 ppm use a multi point calibration with 3 to 5 different standard concentrations Above 0 5 ppm concentration quench ing occurs and samples should be diluted into the linear range before
27. eadings have changed significantly Recalibrate when the accuracy becomes unacceptable for your study TD 700 Calibration Options There are three calibration procedures available on the TD 700 Simple Mode Raw Fluorescence Calibration Multi Optional Mode Raw Fluorescence Calibration Multi Optional Mode Direct Concentration Calibration 1 Simple Mode Calibration In the Simple Mode calibration procedure the instrument range is automatically set based on the typical standard or sample chosen You cannot manually adjust the instrument range when calibrating in this mode The chosen sample will be automatically set to 5096 of the maximum value that can be accurately read by the instrument 500 out of 1000 Also there is no option to read and subtract a blank during this calibration procedure 2 Multi Optional Raw Fluorescence Mode In the Multi Optional Mode Raw Fluorescence calibration you may manually adjust the instrument range by changing the default sample value of 500 to another number from 100 to 950 If you assign a value higher than 500 to your chosen sample you will decrease the maximum sample concentration that can be read and increase the instrument s sensitivity and resolution If you assign a value lower than 500 you will increase the maximum concentration that can be read and decrease the instrument s sensitivity and resolution TD 700 Laboratory Fluorometer Operating Manual Page 19 Direct Concentration Calibrat
28. ed To View the Last Calibration To Abortthe Calibration DOWD Reading Samples A Reading Samples B Resetting Blank to Zero C DataStream TD 700 Laboratory Fluorometer Operating Manual 11 13 18 24 Page 3 APPENDICES APPENDIX 1 Before Calibration or Reading Samples 30 Materials Needed When Handling Samples Linear Range and Quenching Figure 1 Linearity Calibration Curve and Quenching Temperature Considerations Positioning Samples Data Quality OW D gt APPENDIX 2 The Calibration Printout 33 Printout Capability with a Printer or Computer Sample Reading with a Printer or Computer Calibration Printout Using the Direct Concentration Calibration Procedure Calibration Printoutin the Multi Optional Raw Fluorescence Calibration Calibration Printout Using the Simple Mode mo oum APPENDIX Alarms amp Diagnostics 36 A AlarmScreen B Diagnostic Screens APPENDIX 4 Troubleshooting 38 APPENDIX 5 Data Collection 8 APPENDIX 6 Sample and Cuvette Adapters 40 A Installingand Removing Sample Adapters B Minicell Adapter APPENDIX 7 Glossary 42 APPENDIX8 Error Messages and Notes 45 A InvalidInput Error Notes 1 and 2 Error3 Error4 Error5 Error6 Home Screen Displays OVER or Negative Numbers Blank Level Error APPENDIX 9 Maintenance Warranty amp Service 47 A Maintenance B Warranty C Obtaining Service APPENDIX 10 Specific
29. el by viewing through the lamp view port If the lamp access panel is removed the power to the mercury lamp will be cut off This safety feature will prevent the user from being exposed to U V light Do not defeat the interlock switch U V light may cause permanent damage to your eyes Check the lamp view port if 1 Theinstrumentis not responding even though the unit is plugged in and the power is on 2 Thereadings are very low unstable or drifting Note After replacing a lamp you must always recalibrate To replace the mercury vapor lamp 1 Unplugthe instrument and remove the lamp access panel by pulling on the two black quick release latches on the rear panel of the instru ment 2 Beforereplacing a lamp make sure the current lamp is fully seated in both the upper and lower lamp sockets Be cautious when remov ing the lamp It may be hot 3 Toremovethe lamp grasp it carefully and turn it about 90 degrees until the prongs line up with the slots on the lamp sockets Slide it out NOTE To avoid breaking the lamp when replacing it make sure it is fully seated At the bottom lamp socket a metal spring exerts pres sure against the lamp be sure this spring is not preventing proper seating before twisting lamp into place 4 Toinstall a new lamp line up the lamp prongs with the slots on the lamp sockets push the lamp in and turn it about 90 degrees so it is firmly seated Make sure both end caps are properly seated
30. erage slope will change depending on the numbers input during calibration There is no best slope value TD 700 Laboratory Fluorometer Operating Manual Page 35 Note that during calibration the maximum you can enter for the Rangeis 1000 However the actual maximum achieved after calibration may be higher depending upon the reading for the Blank and for the High Standard This is a normal condition the instrument will set to the best sensitivity to obtain the Max Range you specified Calibration Printout in the Multi Optional Raw Fluorescence Mode Example only Your numbers will vary Press D from the Setup Cal screen to send the calibration information to the printer or computer Mode Multi Optional Cal Procedure Raw Fluor Units fsu Fluorescent Signal Units the Sensitivity Level 48 designated units for Raw Fluorescence readings RAW Fluor Sample Blank 500 0 RAW Fluor Blk 123 0 Adj Fluor Sample 377 0 Adj Fluor Blank 0 000 An Explanation of the Printout The first four lines of the printout display the TD 700 s current parameters the mode the cal procedure the units and the instrument sensitivity level Following this is the calibration stored in the unit s memory The calibration stored consists of the raw and adjusted sample and blank value if appli cable expressed in raw fluorescence units Calibration Printout Using the Simple Mode Example only Your numb
31. eroing the instrument to subtract blank Calibration Curve See Linearity By using multiple standards during calibration in the Direct Concentration calibration procedure a calibration curve will be obtained which will provide accurate readings even when the readings are nonlinear Dark Current Asmall amount of current flows in a photomultiplier tube even when the tube is operated in a completely dark state This output is called dark current and the magni tude is greatly dependent upon the amount of voltage applied to the photomultiplier tube i e the greater the Sensitivity Factor the larger the dark current Detection Limit May be used to refer to either the minimum or maxi mum concentration that can be read Limits are specific to the fluorescent material filters lamp and sample or cuvette adaptor used DirectConcentration Refers to the actual concentration of the substance being read Direct Concentration is a user selected TD 700 calibration procedure which after calibration provides the actual concentration of samples in the units selected by the user Discrete Sample Averaging A feature of the TD 700 which allows a sample reading to be averaged over a set period of time which mini mizes operator guesswork temperature effects and inconsistent sample handling When lt gt is pressed from the HOME screen after a 7 second delay the signal willbe averaged for 12 seconds and the average displayed for 5 se
32. ers will vary Press D from the Setup Cal screen to send the calibration information to the printer or computer Mode Simple Cal Procedure Raw Fluor Units fsu Sensitivity Level 48 RAW Fluor Sample 500 0 An Explanation of the Printout The first four lines ofthe printout display the TD 700 s current parameters the mode the cal procedure the units and the instrument sensitivity level Following this is the calibration stored in the unit s memory The calibration stored consists of the sample value only expressed in raw fluorescence units TD 700 Laboratory Fluorometer Operating Manual Page 36 APPENDIX 3 Alarms and Diagnostics A Alarm Screen There are alarms built into the TD 700 to warn of an internal instrument malfunction orlow power Table 1 Alarms Alarm Delay Alarm Condition Normal High Voltage HV Bad 3min 10000r 25V 15010850 Low Power Low PWR 1 min lt 40 100 ALM XXX ppm lt ENT gt Setup amp Cal An alarm will be activated if the abnormal condition is in effect for a specified delay period If an alarm is activated ALM will blink on the HOME screen HV Bad To see what alarm is active from the home screen press lt ESC gt For example if the high voltage HV is outside the specified range you will see HV If the HV alarm is triggered HV Bad is seen when lt ESC gt is pressed from the HOME screen contact Turner Designs If the low power alarm is t
33. est tube or cuvette the sample adaptor and Filter Cylinder from the instrument Completely dry all of these items and the optical filters 3 Wipe up any moisture inside the sample chamber 4 Plugin the unit and turn on the power Allow it to run for a few hours until completely dry inside Periodically wipe off the outside of the instrument with a damp cloth Do not use solvents or abrasive cleaners to clean the TD 700 B Warranty Turner Designs warrants the TD 700 Fluorometer and accessories to be free from defects in materials and workmanship under normal use and service for a period of one year from the time of initial purchase with the following restrictions 1 The instrument and accessories must be installed powered and operated in compliance with the directions in this TD 700 Laboratory Fluorometer Operating Manual and directions accompanying the accessories 2 Damageincurredin shipping is not covered 3 Damageresulting from measurement of samples found to be incom patible with the materials used in the sample system is not covered 4 Damageresulting from contact with corrosive materials or atmosphere is not covered 5 Damagefrom sea water and other moderately corrosive materials that are notpromptly removed from the instrument is not covered 6 Damage caused by modification of the instrument by the customer is notcovered 7 Failure of limited life parts such as lamps is not covered TD 700 Laborat
34. f the fluorescence of a sample reads between point 2 and point 3 the slope of the segment that connects point 2 and point 3 segment 3 is used to calculate the direct concentration of this sample The following equation would be used to calculate sample concentration 8 8 8 B 8 Fluorescence Hesponse Measured 8 8 8 Page 25 Insert Hi Std 6 and press lt gt SETTING sens Sens Factor Sensitivity Set Sens Factor XX Reading Hi Std WAIT X Std Conc XXX X T 1 OK 9 Change Std Conc XX X New ENT Insert X Std and press Reading Insert Blank and press ENT Press 0 when value stable XXX Reading Blank Blank XXX Calibration Complete TD 700 Laboratory Fluorometer Operating Manual Page 26 of a sample that had a fluorescence between points 2 and 3 z F F m C Where F is the fluorescence of the sample F is the fluorescence of standard 2 the slope of the line between points 2 and concentration of standard 2 C ToView the Last Calibration From the HOME Screen press lt ENT gt to reach the Setup Cal screen From the Setup Cal screen press lt D gt to send the current calibration information to a printer or computer Press lt ESC gt to return to the HOME Screen 15 Abort Gal D Abort the Calibration Re To abort the calibration press lt ESC gt at any time during the calibrat
35. g Manual Page 46 G Calibration is bad This may occur if the actual value you entered for a Error6 Bad SE standard does not match its fluorescence reading in comparison with one Calibration or more of the other standards For example you put in a standard you say has an actual value of 10 and it reads 150 and you put in another standard you say has an actual concentration of 5 and it reads 200 Press lt 5 gt then 1 to abort the calibration calibrate again using properly diluted and labeled standards H HOME Screen Displays OVER or Negative Numbers 1 OVER If the HOME screen displays OVER and the lid is closed this means that the sample reading is too high for the instrument s light detector to read at the current level of sensitivity i e sensitivity is set too high raw fluorescence reading 1000 If you see OVER check the reading in the Diagnostics sequence If it is 21000 make sure the proper filters are installed Examine the filters for damage freckling of mirrored surfaces cracks etc If you are reading samples of high concentration you can dilute them until you get an on scale reading Or if you wish to reduce sensitivity calibrate the unit again If you are in the Direct Concentration calibration procedure choose a higher Max range conc or a more concentrated high standard If you are using the Raw Fluor Calibration Procedure use a more concen trated standard a
36. ibration in the Simple Mode Similarly a calibration in the Multi Optional Mode Direct Concentration will not affect a calibration in the Multi Optional Mode Raw Fluorescence Thus in effect the TD 700 can store three separate calibrations at the same time one for each mode This also means that when you change modes as each mode is a separate function you must calibrate in the mode you want touse VI Calibration Overview Why Calibrate The TD 700 calibration procedures setthe instrument s sample range concentration range and sensitivity based on a chosen fluorescent standard or sample In addition calibration in the Direct Concentration mode assigns a digital value to a known standard so that subsequent standards or samples can be easily referenced to the original For ex ample during Direct Concentration calibration you can assign a value of 500 to a sample and know with certain confidence that a sample that reads 250 contains half as much analyte as the original sample assuming that the blank reads zero When to Calibrate For greatest accuracy calibrate before running a new batch of samples Recalibrate if the ambient temperature changes by 5 Recalibrate after changing lamps filters cuvette sizes or the analyte you are measuring Verify the need to calibrate by reading a stable known concentration standard immediately after calibration and again every few hours to see if r
37. in until it is flush against the filter Filter Cylinder Assembly E HO Q TD 700 Side View TD 700 Filter Cylinder Assembly TD 700 Laboratory Fluorometer Operating Manual Page 10 TD 700 Laboratory Fluorometer Operating Manual Locate the emission filter to be installed Handle the filter at the edges so as to avoid leaving fingerprints on the filter or wipe off the filter before installing If the filter has one side that is mirrored or highly reflective it should be installed so the mirrored side faces toward the inside of the cylinder toward the sample Insert it mirrored side in if any in the opening marked with the appropriate set letter EM Push the filter in so it rests flush with the back of the opening Then reinsert the o ring and press in with your plastic pen cap until it is flush against the filter Position the Filter Cylinder in the sample chamber so that the align ment mark for the filter set you are using is aligned with the silver alignment mark on the inside rim of the sample chamber Lamp Installation and Removal There are two types of lamps available for the TD 700 the Mercury Vapor andthe Quartz Halogen The Mercury Vapor Lamp The low pressure mercury vapor lamp comes in various types depending onthe application of interest The average life of a mercury vapor lampis 8000 hours The lamp can be checked without removing the lamp access pan
38. ion sequence Press 1 to abort or lt ESC gt to resume TD 700 Laboratory Fluorometer Operating Manual Page 27 NOTES TD 700 Laboratory Fluorometer Operating Manual Page 28 1 DLY XXX units ENT Setup amp Cal AVG XXX units ENT Setup amp Cal END XXX units ENT Setup amp Cal HOME Screen during Discrete Sample Averaging sequence TD 700 Laboratory Fluorometer Operating Manual X Reading Samples A Reading Samples Fill a clean test tube or cuvette your with sample wipe the outside dry and insert it into the sample adaptor in the sample chamber Close the lid Recordthe sample reading Youhavethree different options for recording sample readings a Discrete Sample Averaging to Printer or Computer Discrete Sample Averaging averages the reading of a sample for 12 seconds This feature minimizes operator guesswork when reading samples provides greater accuracy and reduces error from temperature effects Inserting a sample and pressing lt gt from the HOME screeninitiates the following sequence 7 second delay period for the signal to stabilize 12 second averaging period 5 second display of readout The words DLY AVG and END appear on the upper left of the LCD display and alert the user to the instrument s current status After the sequence is complete the display reads END the reading will freeze for 5 seconds and the readout will automaticall
39. ion Direct Concentration Calibration only available in the Multi Optional Mode is a multi point calibration in which up to five standards and a blank are read The software uses these points to set the optimal instrument range and sensitivity and to calculate the direct concentration of unknowns In a multi point calibration the instrumentgenerates a calibration curve for superior accuracy The TD 700 will display the actual concentration of your samples The display units are user selectable and chosen in the setup menu Note Fluorescence measurements are affected by factors such as temperature linearity and instrument drift Before calibrating or running samples for the first time we recommend reviewing Appendix 1 TD 700 Laboratory Fluorometer Operating Manual Page 20 1 Setup 2 Calibration 1 Mode lt Simple gt Multi Optional 1 Setup 2 Calibration Insert Typical Sample lt ENT gt SETTING Sens Sens Factor XX Sensitivity Set Sens Factor XX Sample XXX ENT CH 2 S 5 o 3 9 o Blank Cal Std XXX X 1 Abort Cal lt ESC gt Resume gt g VI L Calibration Simple Mode The Simple Mode calibration procedure is a single point calibration in which a standard is run in order to set the optimal range and sensitivity of the instrument The chosen sample will be set automatically to one half of the maximum value that ca
40. ives the standard a relative value of 500 on a scale of 0 1000 The instrument uses a preprogrammed absolute zero as the O point Multi Optional Raw fluorescence two point calibration standard blank This calibration allows you to use one standard and the option to blank subtract The user can specify the magnitude in relative fluorescence units for the standard on a scale of 0 1000 As an example if you expect your standard is 75 of the full range for the application see application notes on our website for ranges then call the standard 750 RFUs MultiOptional Direct Concentration Multi point calibration up to 5 standards blank This calibration allows you to use up to 5 different standards plus a blank for the calibration You can choose the units of your standards and define the magnitude of the maximum readable level In the end the output can be in direct concentration so no further calculations are needed TD 700 Laboratory Fluorometer Operating Manual Page 15 NOTES TD 700 Laboratory Fluorometer Operating Manual Page 16 TURNER DESIGNS Power up Screen T7 1B 10 96 600 Countdown from 600 1 Setup XXX units lt ENT gt Setup amp Cal Home Sergen Simple Mode DLY XXX units lt ENT gt Setup amp Cal AVE XXX units lt ENT gt Setup amp Cal END XXX units lt ENT gt Setup amp Cal 0 Blank 1 Yes 9 No ppb lt ppm gt Return to home ppt ug ml screen
41. l Screen To access the calibration sequence press 2 from the Setup Cal Screen The Direct Concentration calibration sequence will appear When the TD 700 calls for the maximum range the maximum concen tration you want to read press 1 to accept the current value or 9 to enter a new value Range for reentry 1 1000 Key in the number of standards you will be using 1 to 5 concentra tions Press ENT When the TD 700 calls for the HiStd Conc press 1 to accept the current value and advance to the next screen Or press 9 to change the value Key in the actual concentration of the highest concentration Standard Hi Std you are using and press ENT The numerical range is 0 1 to 1000 If you are using only 1 standard IT MUST be within the linear range for accurate results see Appendix 1 fora discussion of linear range TD 700 Laboratory Fluorometer Operating Manual 6 Fillaclean test tube or cuvette with the Hi Std highest concentration standard you are using Wipe the outside of the cuvette dry and insert it into the sample adaptor in the sample chamber Press lt gt The unit will adjust sensitivity as shown by the SENS FACTOR to the level appropriate for that standard then read the standard 7 Ifyou are using only 1 standard the TD 700 will prompt you to insert the Blank If you are using 2 or more standards the unit will prompt you to enter the actual concentration of the second standa
42. ly as for Warranty Service above If our service department can assist you by phone or correspondence we will be glad to at no charge Repair service will be billed on a basis of time and materials A complete statement of time spent and materials used will be supplied Shipment to Turner Designs should be prepaid Your bill will include return shipment freight charges Address for Shipment Turner Designs 845 W Maude Ave Sunnyvale CA 94085 TD 700 Laboratory Fluorometer Operating Manual Page 49 APPENDIX 10 Specifications and Features Sample Adaptors 25 mm sample adaptor accommodates 25 x 150 mm test tubes a 13 mm sample adaptor for 13 x 100 mm test tubes and an adaptor for 10x 10 x 45 mm square cuvettes A Range Extending Adaptor with variable apertures for 10 x 10 x 45 mm square cuvettes is also available Sensitivity Less than 20 picograms ml FITC Power External power supply 100 240 VAC 1096 ofthe nominal voltage Max 30 watts Dimensions 23 5 cm W x 28 cm D x 21 cm H 9 25 W x 11 D x 8 25 Weight 5 9 Kg 13 Ibs Operating Temperature 41 104 5 40 C Display 16 x 2 character LCD 3 86 x 86 9 8 cm x 2 18 cm Keypad 4 x 5 keys 3 x 2 7 7 62 cm x 6 86 Data Output 100 ASCII format through a 9 pin RS 232 serial cable at 9600 baud Filter Cylinder Accommodates eight 1 inch round filters four excitation and four emission Software Menu driven microprocessor cont
43. n be accurately read by the instrument You cannot manually adjust the instrument range and sensitivity during this procedure Also there is no option to read and subtract a blank If you do not require these capabilities you may wish to use the Simple Mode calibration as it is expedient and straightforward 1 To choose the Simple Mode press ENT from the HOME screen press 1 for Setup then 1 again for Mode Use lt lt gt gt to choose the Simple Mode Press lt ESC gt twice to return to the Setup Cal Screen Press 2 from the Setup Cal Screen The Simple Mode calibration sequence will appear Fill a clean test tube or cuvette with a sample that is about half the maximum concentration you wish to read You do not need to know the exact concentration you are using it to set the optimal instrument range and sensitivity Wipe the outside of the test tube or cuvette dry and insert itinto the sample adaptor in the sample chamber The TD 700 will now set sensitivity as indicated by the SENS FACTOR sensitivity factor so that the sample you inserted will read 500 half of maximum on the HOME screen Upon finishing the sensitivity adjustment the screen prompts you to acknowledge the value set by pressing ENT Then calibration data will automatically printout to a printer or a computer and return you to the HOME screen See Appendix 2 for details regarding the calibration printout Errors After reading the sampl
44. n the Simple Mode or Multi Optional Mode Raw Fluor Calibration Procedure to see the raw fluorescence for the standard and the blank as set during the last calibration from the Calibration amp Setup menu press 9 and they will appear on the display Operation Hour Oper Shows how many hours the instrument has been operating Power level When at 100 indicates power supply is functioning correctly TD 700 Laboratory Fluorometer Operating Manual Page 38 APPENDIX 4 Troubleshooting Symptom Power ON but readings stay at zero Power switch ON but no power Display reads OVER Display reads negative Drifting readings Erraticreadings Low readings notenough resolution TD 700 Laboratory Fluorometer Operating Manual Solution Check the lamp change if necessary Make sure the sample lid is closed tightly Make sure unit is plugged in securely check power from the wall socket Make sure power supply is functioning Sample reading is too high for the instrument to read at the current sensitivity level Dilute samples to get on scale reading Or recalibrate and reduce sensitivity Reading for Blank is less than zero due to drift reading for samples are less than Blank Reset Blank to Zero or recalibrate The instrument is open loop and readings will drift over a period of time recalibrate to cure Instrument readings will fluctuate somewhat due to instrument noise e
45. nd or chose a lower value for the sample setting if you are inthe Multi Optional Mode NOTE On rare occasions you might see OVER This means that the raw fluorescence reading is less than 1000 This might happen if you calibrated with an extremely high Blank Donotleave the instrument reading OVER for an extended period of time several minutes 2 Negative numbers If the HOME screen is displaying a negative number it means that the sample is reading less concentrated than the Blank This might occur in a case where you calibrated with a high Blank or it might occur if the Blank is reading negative due to instrument drift To cure if you are in the Raw Fluorescence Calibration Procedure you can use the Reset Blank to Zero function Or you can recalibrate Error Blank L Blank Level Error Itoo high If the blank level is too high when the user tries re blank the instrument from the HOME screen this error may occur Most likely this error is a result of using the wrong solution to blank with To remedy use the correct blank solution If the error persists recalibrate TD 700 Laboratory Fluorometer Operating Manual Page 47 APPENDIX 9 Maintenance Warranty amp Service A Maintenance If possible do not spill liquids into the sample chamber However there is adrain plug on the bottom of the instrument underneath the cuvette adaptor If there is a spill 1 Unplugtheinstrument 2 Remove the t
46. nstrument for drainage if there is a spill in the sample compartment Lamp Viewport The mercury vapor lamp access panel has a lamp viewport that allows the user to observe whether the lamp is lit without opening the door 2 with Halogen Lamp T D 700 with Mercury Vapor Lamp Rear View Rear View 10 Optical Filter Installation and Removal The TD 700 Fluorometer musthave two different optical filters to operate correctly an excitation filter and an emission filter TD 700 filters are one inch diameter round colored glass or highly reflective glass The filters are mounted in the Filter Cylinder using flexible rubber o rings The Filter Cylinder is accessed through the sample chamber The Cylinder can hold four different excitation emission filter combinations and is labeled on one rim A and B and labeled on the opposite rim C and D Each of these letters identifies a filter application set Once the filter sets are installed properly in the cylinder changing from one application to another requires a simple repositioning of the cylinder and possibly a lamp change To remove or install optical filters in the Filter Cylinder a Remove any test tubes or cuvettes and sample adaptors from the instrument Then grasp the Filter Cylinder by the rim and pull straight up and out of the unit b Locate th
47. ory Fluorometer Operating Manual Page 48 C Obtaining Service Warranty Service To obtain service during the warranty period the owner shall take the following steps 1 Write or call the Turner Designs service department and describe as precisely as possible the nature of the problem 2 Carry out minor adjustments or tests as suggested by the Service Department 3 If proper performance is not obtained ship the instrument prepaid to Turner Designs with a statement of shipping charges The instrument will be repaired and returned free of charge along with a check to cover shipping charges for all customers in the contiguous continental United States For customers outside of the contiguous continental United States and who have purchased our equipment from one of our authorized distributors contact the distributor If you have purchased direct contact us We will repair the instrument at no charge but we will not pay for shipment documentation etc These charges will be billed at cost NOTE Under no conditions should the instrument or accessories be returned without notice Prior correspondence is needed a To ensure that the problem is not a trivial one easily handled in your laboratory with consequent savings to everyone b To specifically determine the nature of the problem so that repair can be rapid with particular attention paid to the defect you have noted Out of Warranty Service Proceed exact
48. out grab the bulb with a tissue and pull it out of the assembly Using the same tissue insert a new bulb orientation does not matter Plug the power jack back into the main instrument and replace the lamp access panel Push in the black quick release latches to snap the panel in place Turn on the power Check to ensure the lamp is lit by viewing it through the fan opening Also make sure the fan is operating If the fan is not operating check the connector plug You must recalibrate if you replace the lamp TD 700 Laboratory Fluorometer Operating Manual Instrument Parameters Firmware Instrument parameters are setthrough the TD 700 firmware interface Firmware screens are called up using the keypad To see how thescreens fit together see the firmware flowchart on pages 16 amp 17 For details about alarms and instrument diagnostics see Appendix 3 Section A and Section B Power up Screen When the unit is first turned on a screen appears showing the title software version and a 600 second 10 minute countdown Once the 600 second countdown expires the HOME screen will appear The countdown allows the instrument to warm up adequately before measurements begin For optimal stability and accuracy it is recommended that you allow the instrument to complete the countdown period For best results leave the instrument turned on in your laboratory If you wish to bypass the countdown period press lt H gt ESC or E
49. please referto the appropriate Material Safety Data Sheet MSDS for information regarding handling When using potentially hazardous chemicals please use closed cap test tubes whenever possible and limit exposure If closed cap test tubes are not available Parafilm should be used to prevent the spillage of volatile or other potentially hazardous chemicals Chemicals should be used in accordance with local regulations and used ina well ventilated area Do not defeat the lamp interlock switch UV light may cause permanent damage to your eyes D Definition of Symbols Direct Current 12 volts D C Easily touched higher temperature parts Caution Read instruction manual and referto warning text gt E TD 700 Laboratory Fluorometer Operating Manual Page 7 ll Hardware Overview A TD 700 Quick View Diagram Alignment Mark Alignment Mark Filter Cylinder Sample Adapter Silver Alignment Mark Sample Chamber Display Keypad d AC TD 700 Laboratory Fluorometer TD 700 Filter Cylinder and 25 mm Round Sample 10 mm Square Cuvette 13 mm Sample Adapter Adapter Adapter TD 700 Laboratory Fluorometer Operating Manual Page 8 T7 14 08794 2 888 TD 700 Front View 4 b Q TD 700 Filter Cylinder Assembly TD 700 Sample Adapters 13 mm 25 mm and 10 mm TD 700
50. rd 2 Std as in step 5 and to insert the second standard 2 Std as in step 6 then the third 3 Std etc Use a clean rinsed test tube or cuvette and insert the next standard press lt gt then ENT when finished 8 Whenallthe standards have been run the TD 700 will prompt you to insert the Blank Fill a clean test tube or cuvette with the Blank wipe the outside dry and insert it into the sample adaptor in the sample chamber Press ENT 9 Wait for the Blank reading to stabilize then press 0 The TD 700 will read the Blank then automatically return to the HOME screen Calibration data will printout automatically if connected to a printer or computer See Appendix 2 for details regarding the calibration data printout How Sample Concentrations will be Calculated As you have read the Direct Concentration calibration procedure accepts multiple standards and one blank The concentration vs fluorescence plot of both the standard and the blank will result in something close to a straight line assuming the standards are within the linear range of the assay and instrument If you draw a straight line from point to point on the calibration plot and record the slope of the line between each point you can use these slope values to calculate to sample concentrations For example the graph below displays a plot of five standards and a blank Drawing aline between each point will give you five different linear segments I
51. riggered check the power at the source and the power supply Refer to the Diagnostic Screens next section for definitions B Diagnostic Screens There are diagnostic screens which show the status of internal fluorometer functions To access these screens from the HOME screen press 8 then ENT to see the next screen in the sequence Sen 96 XX EE UNS Definitions Oper XXX Hrs Sen 96 Sen is also referred to as Sens Factor in the software and Power level 100 Sensitivity Level on the printout All three terms indicate the fluorometer s sensitivity on a scale of 1 to 100 The Sen is directly proportional to the voltage on the photomultiplier tube The higher the number the greater the photomultiplier tube voltage the greater the sensitivity and typically the greater the noise of the readings TD 700 Laboratory Fluorometer Operating Manual Page 37 Raw The raw fluorescence signal output from the instrument e light detector the photomultiplier tube This is the output the TD 700 uses in conjunction with the actual value entered for the standards and the fluorescence signal for standards and blank to arrive at the readout on the HOME screen It can be used to diagnose problems with the unit NOTE To see the raw fluorescence for the standard s and the blank as set during the last calibration from the Calibration amp Setup menu press D to print or send to a computer NOTE If you calibrated i
52. rolled Detector Factory installed photomultiplier tube Stan dard 300 650 nm Optional Red Sensitive 185 870 nm Lamp Quartz Halogen Lamp 20 watts Lamp Life 2000 hours or Low Pressure Mercury Vapor Lamp 4 watts Lamp Life 8000 hours Printer Optional Seiko Printer 16 0 cm W x 17 0 cm D x 6 7 cm H Material UL approved polyurethane Sheet metal back and base Readout Direct Concentration or Raw Fluorescence Environmental Conditions Indoor use only Altitude specification 0 2000m Transient Overvoltages according to INSTALLATION CATEGORY II Pollution Degree 1 and 2 accordance with IEC 664 TD 700 Laboratory Fluorometer Operating Manual Page 50 Maximum Relative Humidity Calibration Operating Mode Blank Discrete Sample Averaging Warranty Approvals TD 700 Laboratory Fluorometer Operating Manual 75 for temperatures up to 31 C decreasing linearly to 50 relative humidity at 40 C Multi point Calibration for Direct Concentration Measurement or Single Point Raw Fluores cence Calibration Simple for the inexperienced user or Multi Optional forthe advanced user Reads and subtracts Blank Sample readings can be averaged to improve accuracy 7 second delay 12 second signal averaging 5 second display readout One year warranty CE TUV and UL Page 51 APPENDIX 11 Principles of Fluorescence Fluorescence is a physical property of certain
53. rough and excite the sample solution Cuvette The cuvette or sample cell contains the sample of interest The cuvette material must allow the compound s absorption and emission light energy to pass through for example compounds that fluoresce below 300 nm require a quartz cuvette The emission light energy will be emitted from the sample solution at a right angle to the incident light energy The size of the sample cell will also effect your measurement The greater the diameter of the cuvette the lower the concentration that can be read Emission Filter Stray light and any background material fluorescence will also be emitted from the sample solution The emission filter is chosen to screen out these components allowing only wavelengths of light specific to the compound to pass through Light Detector The TD 700 light detector is a photomultiplier tube The light allowed to pass through the emission filter the wavelengths of light specific to the compound are detected by the photomultiplier tube The light intensity which is directly proportional to the compounds concentration is then registered as a digital readout TD 700 Laboratory Fluorometer Operating Manual Page 53 APPENDIX 12 Instructions for Using the Solid Standard with the TD 700 Fluorometer 1 Calibrate your instrument with a known standard as you usually do per sections VII IX 2 Removecalibration standard and any cuvette adaptors from instrument 3 F
54. sample measurement If the sample used is too concentrated or too dilute the instrument may not be able to reach the target sensitivity In these cases an error message will appear indicating that based on the calibration the unit has reached its sensitivity maximum or minimum By pressing lt ENT gt you tell the instrument to accept the maximum or minimum sensitivity value It is recommended however that you adjust your sample concentrations to fall within range and then recalibrate To view the last calibration data set press ENT from the HOME screen then 9 to view your blank and calibrating standard value Press H to return to the HOME screen To print the last calibration data set press ENT from the HOME screen then D Press lt H gt to return to the HOME screen To abort the calibration press lt ESC gt at any time during the calibration sequence Press 1 to abort or lt ESC gt to resume Page 23 Insert Blank and press ENT Press 0 when value stable XXX Reading Blank WAIT Blank XXX Blank X X Cal Std XXX X 1 Abort Cal lt ESC gt Resume TD 700 Laboratory Fluorometer Operating Manual Page 24 Calibration Multi Optional Direct Concentration 1 Setup 2 Calibration 1 3 Units 2 Cal Procedure Simple lt Multi Optional gt 1 Mode 3 Units 2 Cal Procedure Raw Fluor lt Direct Conc gt 1 Setup 2 Calibration
55. specially at high sensitivity and when reading low concentrations If readings are very erratic check the lamp and opticalfilters for damage freckling Sensitivity was not adjusted adequately during calibration recalibrate Or make sure the proper lamp and filters are being used Section To See Section IV Appendix 8 H Appendix 8 H Section VII Section VIII Section IX Section Ill Section IV Section VII Section VIII Section IX Section III Section IV Page 39 APPENDIX 5 Data Collection You can use various communications software programs to import data to your computer The signal is sent from the TD 700 in ASCII format 1 Connectthe 9 pin connector that extends from the TD 700 to the serial communications port of your computer COM1 or COM using a 9 pin serial cable You may need a 9 to 25 pin serial adaptor Obtain this at your local computer store 2 In Window s OS locate the HyperTerminal program in the Accessories directory and double click on the icon to open it If operating in MAC OS use a comparable terminal program to capture the ASCII data 3 Give the New Connection a name such as TD700 4 In the Phone Number window choose Direct to COM 1 or Direct to COM 2 depending on which COM Port that the instrument is connected to This can be the hardest part of the procedure Different computer manufacturers use different conventions 5 Set the Port Settings to the following parameters Bi
56. taking readings Or you can switch to a smaller size test tube or cuvette smaller pathlength and increase the linear range Linearity may be checked by diluting a sample 1 1 or some other conve nient ratio If it is linear the reading will decrease in direct proportion to the dilution Ifthe reading goes up concentration quenching you are beyond the range for a calibration curve If the reading does not decrease in direct proportion to the dilution you are in the range for a calibration curve but still beyond the linear range 4 Concentration Quenching is Observed Range in which Multi Point Calibration is needed 60 80 100 Concentration Figure 1 Linearity Calibration Curve and Quenching Temperature Considerations Fluorescence is temperature sensitive As the temperature of the sample increases the fluorescence decreases For greatest accuracy read blank standard and samples at the same temperature The TD 700 Fluorometer does not compensate for temperature changes NOTE The unit s lamp will heat samples if left in the instrument Thus it is recommended that you use the instrument s Discrete Sample Averaging feature to ensure that samples are read after they have been in the instrument for the same period of time and are at approximately the same temperature TD 700 Laboratory Fluorometer Operating Manual Page 32 E Positioning Samples Sample cuvettes often will give slightly
57. ts per second 9600 Data Bits 8 Parity none Stop Bits 1 Flow Control Xon Xoff 6 To store the data to a file choose the Transfer menu and select Capture Give the captured file a name 7 Stop the Capture when you are done by selecting the Transfer menu and then by selecting Capture Stop 8 Access your data using Excel Word or other programs by opening the file you saved in step 6 TD 700 Laboratory Fluorometer Operating Manual Page 40 APPENDIX 6 Sample and Cuvette Adapters Test tube and cuvette size apertures and the sample cell path length all affect the sensitivity of the instrument In general the greater the diameter of the test tube or cuvette the greater the path length the more sensitive the readings and the lowerthe linear range The smaller the window and path length the less sensitive the readings You must recalibrate the instrument if you change cuvette adapters or if you change path lengths or attenuators A Installing and Removing Sample Adapters Available for the TD 700 are a variety of sample adapter to cater to the sample size needs of researchers These are 25 mm test tube adapter P N 7000 982 standard with TD 700 model 7000 000 13 mm test tube adapter P N 7000 981 standard with TD 700 model 7000 000 10x10 mm square cuvette adapter PN 7000 988 standard with TD 700 model 7000 009 Minicell adapter P N 7000 951 fits inside 10x10 cuvettes adapter P N 7000 009
58. vity of the instrument usually expressed in powers of 10 Linear Range The concentration range in which standards correlate with a straight line Usually expressed as a concentra tion maximum such as 100 ppb Noise The amount of fluctuation in the instrument s measure ment due to the instrument or detector itself Optical Path Length length of sample through which light passes This is defined by the cuvette size and adaptor used Quenching A phenomenon which occurs when the fluorescence measurements decrease even though analyte concen tration is increasing Raw Fluorescence Refers to the relative fluorescence of a substance being read rather than the actual concentration Raw Fluorescence is a user selected TD 700 calibration procedure which after calibration provides the fluores cence intensity of samples relative to the standard and to each other TD 700 Laboratory Fluorometer Operating Manual Page 44 Resolution Sens Factor Sen 96 Sensitivity Level Sensitivity TD 700 Laboratory Fluorometer Operating Manual Resolution is related to sensitivity but refers to the detail which can be read When the instrument sensitivity level is high lower concentrations can be read with ease high resolution but higher concentra tions can not be read on scale When the sensitivity is low higher concentrations can be read but the detail is not as good at low concentrations low resolution These
59. ws terminal see Appendix 5 3 Press D or lt gt from the HOME screen and the readout currently displayed will be sent to the computer with an index number B Sample Reading with a Printer or Computer When D or is pressed from the HOME Screen the TD 700 sends out an index marker 1 999 the concentration readout of the sample and the Raw Fluorescence all in ASCII format Note that the column headings will appear only before the first index and the index will begin again from 1 every time you calibrate or turn off the instrument An example of the indexing feature is illustrated by the left column in the sample printout below In the Direct Concentration Calibration Procedure for example only Samples Conc Raw Fluor 1 32 01 42 80 2 16 00 21 40 3 8 002 10 70 For the Simple and Raw Fluorescence modes for example only Samples Raw Fluor 1 42 80 2 21 40 3 10 70 TD 700 Laboratory Fluorometer Operating Manual Page 34 C Calibration Printout Using the Direct Concentration Calibration Procedure Example only Your numbers will vary Press D from the Setup Cal screen to send the calibration information to the printer or computer Mode Multi Optional Cal Procedure Direct Conc Units ppb Sensitivity Level 48 Current Calibration Entered Max Range 220 00 Calibration Standard Summary Sid Conc Raw Fluor BLK 0 0 Std 3 1 50 6 706 Std 2 6 20 29 09 Hi Std 50 00
60. y be sent to your computer or printer The letters disc will appear next to the reading on the printout to indicate that the discrete sample averaging feature was used on this measurement Read Samples from Display Your second option is to read samples directly from the display without using the Discrete Sample Averaging function Allow the reading to stabilize then press D to send the reading to a computer or printer or write down the result Data Stream Instantaneous readings are reported out to a printer or computer at a preset interval See Section X C for futher details 3 Rinse the cuvette 3 times preferably with the sample you wish to measure Repeat steps 1 through 3 until all samples are read Note Do not leave highly concentrated samples in the instrument for an extended period of time Unstable readings may result Page 29 Resetting Blank to Zero This feature allows you to reset the blank to zero from the HOME screen when in the Multi Optional Mode Although running a calibration is the best method to minimize sample error due to instrument drift a quick and easy way to reduce the effects of dark current and blank fluorescence is to autozero or reset the blank to zero from the HOME screen However be sure to run at least one calibration before using this feature Resetting the blank to zero from the HOME screen is quite useful if you wish to read multiple sets of blank and relative sample readings an

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