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OPERATOR`S MANUAL

1. This menu is analogous to that for the AO1 range described above Wndw Pressing the Wndw button brings up the following screen Si o e Ti me Wi n do w Set Hel p Show Bac k 1 Set Pressing the Set button allows you to select the growth rate time window allowed values are Imin 2min 4min 8min 16min 32min 8min is the default setting This setting determines the range of time over which fitting is performed when estimating the exponential growth rate constant 2 Show Selecting the Show key brings up a screen in which the current setting is displayed for 3 sec Communication Select Comm BE2100 base units only Pressing the button below Comm brings up the following screen C e E i c ati om le u in RS 28 p2 Show Baek m S m US B 73 h i ii Selecting USB or RS232 results in that communication port becoming active The default setting is USB When USB is selected RS 232 communication will still function as long as the USB cable is not also plugged in When RS232 is selected USB communication is disabled Show Selecting the Show key brings up the fo
2. STEP 3 Press the arrowhead beneath the word Ave The display should now look like 11 Ti me Constant Sel ect See t el p Show Bac A STEP 4 Press the arrowhead beneath the word Help A brief description of the function flashes onto the screen after which the screen reverts to that shown above in Step 3 STEP 5 Press the arrowhead beneath the word Show The currently selected time constant is shown along with the default time constant STEP 6 Press the arrowhead beneath the word Set The display should now look like Sel ebct Ti me Constant S e c 0 Se Mor Bac Pressing the arrowhead below the word More would lead you to menus with other choices for time constants Time Constants between 0 seconds and 8 minutes can be selected Note Use of the 0 sec time constant is generally not advised except when very fast sensor response is required or as a temporary measure such as when initially positioning the sensor on a crowded bioreactor vessel When the time constant is set to 0 seconds digital filtering is turned off and only analog filtering with a time constant of approximately 3 seconds is applied to the reported sensor data STEP 7 Press the arrowhead beneath 30Sec The screen will briefly show the new value that has been set
3. 12 13 Using the Voltmeter measure the DC voltage across the 500 Q resistor Record this as V low Replace the low calibration cup with the high calibration cup Secure the high calibration cup against the sensor as detailed in step 4 above The biomass reading on the bottom line of the base unit should now be greater than 100 Bug Units Using the Voltmeter measure the DC voltage across the 500 Q resistor Record this as V high Return the sensor averaging time constant range baseline correction and user calibration settings to their prior settings Convert V low and V high from voltages V to currents mA by multiplying by 2 Ilow 2 V low high 2 V high Use I low and I high as calibration inputs for your analog reading device e g bioreactor controller This step will vary depending on the control software you are using but typical linear calibration inputs are offset and span values in units of current and corresponding biomass units The table below shows the correspondence between offset and span and the values you just measured Table 1 Example Linear Analog Calibration Inputs Current mA Biomass Offset I low 0 00 Span I high I low BE2x00 base unit Range setting Note that the Range value you use for the Biomass Span should be the Range setting that you will select during your bioreactor run not the Range setting just used during the calibration procedure
4. Bug Units are generated at the time of sensor manufacture by linearly scaling the measured optical 850 nm reflectance from a calibration solution of Saccharomyces cerevisiae in a 14L glass fermentor to Optical Density OD measured at 600 nm in a 1 cm cuvette in a spectrophotometer Perkin Elmer model Lambda 9 with appropriate dilution Absorbance lt 0 5 and scaling by the dilution factor Due to the differences between the OD measured between different commercially available spectrophotometers and the cell size dependence of the correlation between biomass and optical scattering see the Theory of Operation section for further details when an absolute reading of biomass is needed it is recommended to calibrate the BE2100 sensor to the particular organism you are growing using your preferred reference biomass method e g dry cell weight OD etc The high linearity of the BE2100 sensor makes performing such calibrations quite simple see User Calibration However in many applications where only measurements of the growth rate or growth trend are needed it is unnecessary to calibrate the BE2100 sensor and measurements in Bug Units will suffice As with spectrophotometry correction for the baseline scattering by the media alone is often useful in distinguishing the biomass from the media As described in the following section a baseline correction function for the BE2100 sensor is provided for this purpose E
5. Overall Length without connectors 13 2cm 5 2 Overall Height without feet Base Unit Environmental Operating Temperature 8 2m 3 2 0 to 40 C 30 to 100F Operating Environment BE 2400 FN D AS 2 N A ENSOR SENSOR ENSOR 3 SOR 4 64 dry location use only S N 240003 uss RS 233 4 BE USB Features and Specifications User Interface Software Features CD included with BE USB Real time graphical and numerical display of sensor data Event marking on graph both pre defined and user defined Baseline setting and subtraction User calibration of sensor output to any reference units Access to all sensor settings User Interface Software Requirements Windows XP Vista 7 8 32 or 64 bit Operating System Minimum of 256 MB of RAM Minimum 200 MB free hard disk space 1024 by 768 resolution or higher video adapter Available USB or 9 pin RS 232 COM port Microsoft compatible mouse CD reader required only at time of software installation BE USB Electrical DC Power In 5V 90mA Digital Output USB Connectors USB B female and BE2100 sensor female 4 wire threaded USB Communications Cable included Connectors USB A male computer and USB B male BE USB Length Standard 2m 6 Custom up to 15m 50 BE USB Physical Overall Width 2 8cm 1 1
6. Step iic Press the arrowhead beneath the word More 20 Step iii Press the arrowhead below Chk The top line of the display should now read Sensor Check Mode Step iv Press the arrowhead below the word Run The top line of the display should now read Attach Low Cal Cup Step v Once the low calibration cup has been securely attached to the sensor see step 2 above press the arrowhead below the word Ready Step vi The instrument will make 25 measurements lasting about 1 second each on the low calibration cup The standard deviation of the 25 measurements is used to assess stability If the measurements were unstable the message Low Cup unstable RPT will be displayed If this occurs make sure that the front faces of the sensor and calibration cup are clean and dry and that the calibration cup is securely attached to sensor see step 2 above You will not be allowed to proceed to measurements with the high calibration cup until stable measurements on the low calibration cup have been collected Step vii Once stable measurements on the low calibration cup have been collected the message Continue to Hi Cal will be displayed Press the arrowhead beneath the word Yes The top line of the display should now read Attach High Cal Cup Step viii Replace the low calibration cup with the high calibration cup Make sure the cup is clean and securely attached to the sen
7. If you change the range setting on the BE2x00 base unit you will also need to update the calibration in your control software Also the table above assumes that baseline correction will be applied to the BE2100 sensor result so that when media alone is measured the biomass reading is zero If baseline correction is not applied and the biomass reading for media alone is not zero you should use this baseline reading as the Biomass Offset and subtract this value from the span However baseline correction via the BE2x00 instrument is generally recommended since this can be applied without having to update the calibration inputs for your control software 91 Appendix V Trouble Shooting Sensor Trouble Shooting Observation Disagreement between BE2100 sensor and offline reference measurement Possible Causes 1 Incorrect calibration selected 2 Incorrect baseline 3 Interference from objects within the bioreactor 4 Cell lysates are contributing significantly to the measured optical reflectance Suggested Remedies 1 See User Calibration 2 Collect a baseline reading on the media alone prior to inoculation See Setting the Baseline 3 See Setting up for a Bioreactor Run Step 4 4 See Principles of Operation Sensor Check test failed 1 Incorrect calibration cups were used 2 Dirt on window or calibration cups 3 Laser aging 4 Laser burned out due to excessive temperature e g
8. LSB MSB 0x55 Quadratic Coeff Cubic Coeff 4 Bytes 4 Bytes LSB MSB LSB MSB MinX MaxX Transform Method 4 Bytes 4 Bytes 1 Byte LSB MSB LSB MSB the coefficients appear as a continuous message this table has been formatted to fit on the page Sending an empty packet U command will return the u message displaying the currently set array Key Offset Linear Coeff 1 Byte 4 Bytes 4 Bytes w Ox75 LSB MSB LSB MSB 87 Quadratic Coeff Cubic Coeff 4 Bytes 4 Bytes LSB MSB LSB MSB Minx MaxX 4 Bytes 4 Bytes LSB MSB LSB MSB the coefficients appear as a continuous message this table has been formatted to fit on the page 13 V 0x56 Sensor Embedded Software Version This command reads the version of the embedded software that exists in the sensor The data field is comprised of a 32 bit floating point value The only format for the V command is an empty V command Key 1 Byte V 0x56 Sending an empty packet V command will return the v message displaying the current setting Key Sensor S W version 1 Byte 32 bits y 0x76 LSB MSB 14 W 0x57 Growth Rate Window The W command determines the time window in seconds to be used when estimating the
9. Note The BE2100 is an optical instrument and will only operate on transparent cylindrical glass bioreactors or bioreactors with a glass viewing port that makes contact with the fermentation solution Configuring the Power Adapter for BE2100 and BE2400 Base Units International Version only Note When sensor connectivity is provided through a BEIUSB adapter power is provided to the sensor via the USB connection ie no separate power adapter is required oe y aie al Continental North American B Europe Plug Plug Adapter UK Ireland Australia New __ gt Plug Adapter Zealand Plug Adapter Temporary Plug Insert International Power Adapter Plug Adapter Release Switch STEP 1 Slide the Plug Adapter Release Switch into the release position towards the power cord and remove the temporary plug insert STEP 2 Select the Plug Adapter appropriate for the country in which the instrument is to be used and insert it while holding the Plug Adapter Release Switch in the release position STEP 3 Release the Plug Adapter Release Switch and check that the Plug Adapter is held securely in the power adapter Connecting the Sensor and Base Unit or BEJUSB Adapter STEP 1 Connect the sensor cable s to the rear of the Base Unit or to the sensor connector on the BEIUSB adapter Make sure the 4 pins on the sensor connector match up with the 4 holes on the Base Unit or BEIUSB connector Screw the connector in
10. The new value is immediately saved into sensor memory and will be recalled even if you switch the instrument off and on again The screen then returns to the 2 Configuration menu as shown above in Step 2 STEP 8 Press the arrowhead beneath the word Ave again The display should now look like Ti mie Const aint Stel ect Sle tt Hel p S h o w Bac A A A 4 STEP 9 Press the arrowhead beneath the word Set The display should now look like Sel ect Ti me Constant sec 0 Se Mor Bac A A STEP 10 Press the arrowhead beneath the word More The display should now look like Sel ect Ti me Constant 1Mi 2 Mi Mor Bac A STEP 11 Press the arrowhead beneath 2Min or select whatever time constant is most appropriate for your measurement conditions The screen will briefly show the new value that has been set The new value is immediately saved into sensor memory and will be recalled even if you switch the instrument off and on again The screen then returns to the 2 Configuration menu as shown above in Step 2 STEP 12 Press the arrowhead beneath the word More Pressing the arrowheads beneath AO1 and AO2 would lead you into sub menus that would allow you to set paramet
11. accurately If the sensor is operated during conditions where the OD of the fermentation is below the range of the internal Bug Units calibration less than 0 001 Bug Units the BE2100 will post the following error message Below Range This error is automatically resolved when the OD of the fermentation reaches the lowest OD of the calibration 32 If the sensor is operated during conditions where the OD of the fermentation is above the range of the internal Bug Units calibration greater than 1000 Bug Units the BE2100 will post the following error message Above Range The Base Unit will inform you if the sensor has become disconnected Sensor Disconnected 33 PRINCIPLES OF OPERATION The BE2100 sensor employs a near infrared 850 nm laser to non invasively measure back scattering from biomass within liquid cultures The laser is directed through the wall of transparent glass or plastic vessels or through a glass port window of stainless steel bioreactors When the laser light is scattered by the cells or microorganisms it creates a glow ball of monochromatic light The intensity and size of the glow ball is dependent on the biomass within the liquid culture At early stages of growth when the biomass is low the glow ball will be large in size and weak in intensity As the cells or microorganisms grow and divide the density will increase and the glow ball will reduce in size and increase in inten
12. rights e Software Installation and Use You may install and use two copies of the BE2100 software on two different computers e Back up Copy You may make one back up copy solely for archival purposes 2 DESCRIPTION OF OTHER RIGHTS AND LIMITATIONS e Limitations on Reverse Engineering Decompilation and Disassembly You may not reverse engineer decompile or disassemble the BE2x00 software e Rental You may not rent lease or lend the BE2x00 software e Termination Without prejudice to any other rights BugLab may terminate your rights under this EULA if you fail to comply with the terms and conditions of this EULA In such an event you must destroy all copies of the BE2x00 software e Trademarks This EULA does not grant you any rights in connection with any trademarks or service marks of BugLab or its suppliers 57 3 COPYRIGHT All title and intellectual property rights in and to the BE2x00 software are owned by BugLab You may not copy the printed materials accompanying the BE2x00 software All rights not specifically granted under the EULA are reserved by BugLab Do not make illegal copies of this software 58 BE2100 Sensor Specifications Sensor Performance Range of OD Sensitivity 0 to gt 300 OD units OD Accuracy 15 OD 0 5 300 typical RMSE in user calibrated mode 0 1 OD OD lt 0 5 Averaging Time Constant 3 sec 8 min Performance Verification Recalibration calibration cup
13. the USB driver software may not have been installed correctly The following procedure describes how to check and if necessary re install the USB communication driver software This procedures assumes that you have already installed the User Interface software if not first follow the steps in Software Installation 1 Check to make sure that the driver was installed properly by checking the status in Device Manager a Make sure that your BE2x00 instrument is plugged into a USB port on your computer b Press the windows Start button and choose Control Panel 94 2 3 4 5 G Locate and double click on the Device Manager icon d Scroll down the devices to Ports COM amp LPT and view the listed devices by clicking on the windows expansion arrow e One of the listed ports should be USB Serial Port COMx where x is an integer e g USB Serial Port COM8 If no such device is listed skip to step 3 below f Right click on the USB Serial Port and select Properties Click on the General tab The Device status window should be displaying This device is working properly If not skip to step 2 below h Click on the Driver tab The driver that is listed should be FTDI version 2 12 0 0 or higher If it is not proceed to step 2 below If the device is not working properly or the wrong FTDI version is listed uninstall the driver software a Disco
14. you to menus with other choices for time constants Time Constants between 0 seconds and 8 minutes can be selected Note Use of the 0 sec time constant is generally not advised except when very fast sensor response is required or as a temporary measure such as when initially positioning the sensor on a crowded bioreactor vessel When the time constant is set to 0 seconds digital filtering is turned off and only analog filtering with a time constant of approximately 3 seconds is applied to the reported sensor data STEP 7 Press the arrowhead beneath the word 30Sec The screen will briefly show the new value that has been set The new value is immediately saved into sensor memory and will be recalled even if you switch the instrument off and on again The screen then returns to the 3 Configuration menu as shown above in Step 2 STEP 8 Press the arrowhead beneath the word Ave again The display should now look like Tli me Constant Sel ect Sie t Hel p S how Bac A A STEP 9 Press the arrowhead beneath the word Set The display should now look like Al AJ LA ia STEP 10 Press the arrowhead beneath the word More The display should now look like Sel ebct Ti me Constant 1
15. 4 Introduction to the BE2100 Base Unit Functions 5 Introduction to the BE2400 Base Unit Functions 6 Verification of Sensor Performance C Setting up on a Bioreactor 1 Attaching the Sensor Head 2 Operation 3 Baseline Correction D Working with the BE2100 and BE2400 Base Units 1 Analog Output 2 Keypad Lockout 3 Display Scrolling BE2400 only 4 Password Locking and Unlocking 5 Warning Messages 6 Error Messages 7 Principles of Operation BE2x00 Software A Minimum System Requirements B Conventions and Shortcuts C Software Installation D Setting Up and Configuring E Data Acquisition 1 Initiating Data Collection 2 Setting the Baseline 3 Modifying the Data Acquisition Window 4 Recording Events during Data Collection 5 Editing Annotations 6 Simultaneous Data Collection from Mult Sens 7 Terminating Data Collection F User Calibration 1 Collecting Calibration Data 2 Editing Generating and Saving a Calibration 3 Running in Calibrated Mode G Data Viewer 1 Opening Viewing and Resaving Data Files 1 Retrospective Baseline Adjustment 2 Retrospective Calibration Adjustment 10 10 14 19 23 26 27 29 30 31 31 31 32 34 37 37 38 39 42 44 45 47 48 49 49 49 50 52 53 53 55 End User License Agreement Appendix I BE2x00 Specifications Appendix II Descript of Base Unit Display Menu and Funct Appendix III Serial Command Set USB RS 232 Appendix IV Analog Output Calibrati
16. Base is pressed the Baseline Correction mode is entered Bas el i ne Opti ons Omn S t a r S h o B a c i On The button under On toggles baseline correction between On and Off In the On state the baseline is used to correct the value displayed during normal operation Once pressed the Baseline On or Off command is sent to the sensor no further user confirmation is required ii Start Pressing the button under Start begins the baseline data collection process Once pressed the word Start will be replaced with End Baseline data collection continues until either the End or the Back button is pressed If the End button is pressed the new baseline value is displayed on the VED for 3 seconds after which the above Baseline Mode screen is then again displayed The new baseline value is saved in the sensor no further user confirmation is required Bas el i me Se t t i ngeer K IX K xX Bu g Uni t s iii Show Pressing the Show button results in the current baseline value being displayed on the VFD for 3 seconds same screen as if the End button was pressed in step b above If Show is pressed while gathering is in progress the top line of the display will read Sensor Busy iv Ba
17. Base Unit S N 1 Byte 4 Bytes 3 0x33 MSB LSB 89 Appendix IV Example Procedure for Calibrating the Analog Outputs on a BE2100 or BE2400 Base Unit The analog outputs AO on BE2100 and BE2400 base units provide current outputs that are proportional to biomass BE2100 and BE2400 or growth rate BE2100 only The nominal range of the current is 4 to 20 mA However due to component variation the actual minimum and maximum currents vary somewhat between base units Therefore when using the AO outputs it recommended that the actual minimum and maximum currents be measured and used as calibration inputs for the AO reading device such as an analog to digital converter on your bioreactor controller The following example provides a step by step method for measuring the minimum and maximum AO currents Tools needed DC Voltmeter with measurement range of 1 mV to at least 10 V Miniature flat head screw driver blade size 2 5 mm Analog output terminal block 4 or 8 position respectively as provided with BE2100 and BE2400 base units 500 Q resistor 1 precision as provided with BE2x00 base units low and high calibration cups as provided with BE2100 sensors Procedure 1 Set the averaging time constant on the base unit to zero see Introduction to the BE2x00 Base Unit Functions for a step by step procedure 2 Set the Range for the analog output you wish to calibrate to 100 see Appendix II Key Pad
18. Communication section v for further details 3 Turn Off both Baseline Correction and User Calibration and then exit back out of the configuration menu The bottom line of the base unit should now show read in Bug Units indicating that User Calibration is Off and the letter C should not be displayed at the end of the line indicating that Baseline Correction is Off see Appendix II Key Pad Communication for further details 4 Remove all of the magnetic adapter sleeves from the sensor Place the Low calibration cup over the front of the sensor Make sure that the arrow marked on the calibration cup is pointing towards the arrow marked on the sensor Connect the buckle components attached to opposite ends of the strap Cinch the sensor against the calibration cup by pulling on the free end of the strap that protrudes from the buckle Next close the latch on the back of the sensor this step should provide the final pressure needed to firmly seat the sensor against the calibration cup 5 The biomass reading on the bottom line of the base unit display should now read 0 00 Bug Units 6 Using a miniature flat head screwdriver attach a 500 Q 1 precision resistor across the and terminals of the analog output terminal block for the analog output that you wish to calibrate 7 Plug the analog output terminal block into the mating connector at the rear of the base unit 90 8 9 10 11
19. Data This section describes how to collect a calibration file for your BE2100 sensor Your calibration will provide the most accurate results if the positioning of the sensor on the bioreactor and the conditions inside the 49 bioreactor are similar between the calibration run and subsequent runs in which the calibration is applied STEP 1 Run a fermentation as you normally would with the BE2100 sensor properly attached and monitoring Follow the directions in the earlier section of this manual entitled Initiating Data Collection STEP 2 At representative points during the fermentation collect calibration samples Right click over the graph each time a calibration sample is removed You will note a special Annotation type called Calibration Sample Removal Each time you record a Calibration Sample Removal Event the software records the time the event number and the raw sensor output Note that the off line reference value and units can also be recorded now in the Value and Units fields However more commonly the off line values will be entered at some later time as they become available Click on the Add Annotation button Notice that calibration events are marked on the graph in blue Use the event number to keep track of the samples you collect for off line analysis STEP 3 Repeat step 2 until a full calibration set has been collected The calibration file must contain a minimum of one point but we strong
20. Mi 2 Mii Mor Bac A STEP 11 Press the arrowhead beneath 2Min or select whatever time constant is most appropriate for your measurement conditions The screen will briefly show the new value that has been set The new value is immediately saved into sensor memory and will be recalled even if you switch the instrument off and on again The screen then returns to the 2 Configuration menu as shown above in Step 2 STEP 12 Press the arrowhead beneath the word More Pressing the arrows beneath the words Range or Disp would lead you into sub menus that would allow you to set parameters associated with the 4 20 mA analog outputs available on the rear of the Base Unit or the display brightness respectively The analog output provides a signal that is proportional to the signal displayed on the front of the Base Unit when not in the Configuration menus The correspondence between the analog output and the signal value is determined by the range setting available in these sub menus STEP 13 Press the arrowhead beneath the word More Pressing the arrowhead beneath the words Pswd would lead you into a sub menu that allows you to password restrict access to the keypad functions Pressing the arrowhead beneath the word Reset would allow you to reset all parameters to their factory default values STEP 14 Press the arrowhead beneath the word Exit This will exit you out of the configuration m
21. Mounting Device 36 BE2x00 Software Minimum System requirements Windows XP Vista 7 8 8 1 32 or 64 bit Operating System Minimum of 256 MB of RAM Minimum 200 MB free hard disk space 1024 by 768 resolution or higher video adapter Microsoft compatible mouse Available 9 pin RS 232 or USB communications port OF Oe OE The BE2x00 software runs in the LabVIEW operating environment Two separate programs are provided 1 the BE2x00 Virtual Instrument and 2 the BE2x00 Data Viewer The Virtual Instrument software gives you the ability to chart the progress of your fermentation in real time and annotate important events Most importantly the software allows you to calibrate your BE2100 sensor to the units of your choice This calibration can then be written into sensor memory allowing the BE2x00 instrument to run in calibrated mode without being connected to a computer The Data Viewer software allows you to open view manipulate and re save data files that were previously acquired with the Virtual Instrument software The Data Viewer software does not communicate with BE2x00 instruments and can be run at the same time as the Virtual Instrument software Conventions and Shortcuts 1 Bold text is used to indicate menu items and buttons that you may select with your mouse key combinations that you may execute on your keyboard and names of control and indicators on the graphical user interface 2 It
22. Overall Length 5cm 2 Overall Height 2 3cm 0 9 BE USB Environmental Operating Temperature 0 to 40 C 30 to 100F Operating Environment dry location use only 65 Appendix II Description of Base Unit Display Menu and Functions i ii iii Startup screen 1 displayed at startup for 5 seconds B ju g IL ja b L IL C Mio in SW Wie lr x x x x Ix x Startup screen 2 displayed for 5 seconds following o screen For BE2400 mux base units the startup screen will be repeated for each of the up to 4 ports to which a sensor is attached If no sensor is connected screen 2 will display the following and the monitor will check periodically for sensor connection If a sensor is connected subsequently the sequence will start from step l a ii above Pee Pe ee eee eee If the above message is displayed when working with a BE2400 base unit but a sensor is connected the instrument may be attempting to connect to a sensor port that is disconnected Press any of the buttons on the keypad and select Scrol to make the display scroll through all available sensors Or select Lock and then choose an available sensor to which you want to lock the display After the startup timer expires the screen reverts to the Normal Operating screen 1 Line 1 Reserved for error warning messages see below If keypad access is bloc
23. Sending the high value hex FF for a byte will preserve the current setting in that byte Byte values other than 0 1 2 3 4 5 and FF are ignored Key AO1 amp 2 Range 1 Byte 1 Byte 1 Byte R 0x52 AOI AO2 664 99 r Sending an empty packet R command will return the r message displaying the currently set values Key AO1 amp 2 Range 1 Byte 1 Byte 1 Byte r 0x72 AOI AO2 The above description applies only to the standard configuration where a sensor is connected to a BE2100 base unit If a BE2400 multiplexing base unit is used instead only one analog output is available for each sensor AO1 changing the range setting for AOZ2 has no effect If the sensor is connected via a BEIUSB adapter no analog output is available so the range command serves no purpose 10 S 0x53 Sensor Serial Number This command reads the Sensor Serial Number The field data is represented as a 32 bit unsigned integer Sending an empty packet S command will return the s message displaying the currently set values It is also sent automatically when the device is first powered up following the send of the Base Unit or BEIUSB embedded software version data Key Sensor S N 1 Byte 4 Bytes s 0x73 MSB LSB 11 T 0x54 User Calibration Units Null terminated string of up to 11 characters describin
24. USB Vacuum Fluorescent Display VFD brightness level Disp The VFD brightness can be adjusted between 4 levels The default VFD brightness level is 3 Password Protection Pswd Allows the user to set a password and lock others out of changing settings unless they enter the correct password Reset Resets all of the above adjustable parameters to their factory default settings 2 Settings b e are stored in sensor memory and settings a and f j are stored in base unit memory The settings are maintained across power cycling of the instrument vi If any of the Soft keys are pressed during Normal Operation the system configuration menu is activated 1 Scroll Lock BE2400 Base Units only The opening screen of the BE2400 configuration menu provides the option of putting the display into scrolling or locked display mode or proceeding to the sensor specific configuration menus a Scrol The display will sequentially scroll between all available sensors at 5 second intervals 69 b Lock The display will be locked to one particular sensor An additional screen is displayed to allow selection of the sensor to which the display will be locked c More Provides entry into the sensor specific configuration menus An additional screen is displayed to allow selection of the sensor to which the display will be locked 2 Sensor specific settings a Baseline Mode Base If the button below
25. arrowhead beneath the word Exit would return you to the normal display screen STEP 2 Press the arrowhead beneath the word More The display should now look like Conf i gurati on Menu 3 Ch k Av e Mor E x ji A Pressing the arrowhead below the word Chk would lead you into the Sensor Check function This function allows you to verify sensor performance and if necessary update calibration coefficients using measurements made on the Low and High calibration cups STEP 3 Press the arrowhead beneath Ave The display should now look like Ti mie Constant Sel ect Sie t el p Show Bac A STEP 4 Press the arrowhead beneath the word Help A brief description of the function flashes onto the screen after which the screen reverts to that shown above in Step 3 STEP 5 Press the arrowhead beneath the word Show The currently selected time constant is shown along with the default time constant STEP 6 Press the arrowhead beneath the word Set The display should now look like 16 Sel ect Ti me Constant Sec 0 S e Mor Bac A Pressing the arrowhead below the word More would lead
26. data is read from the sensor displayed in graphical form and saved to file data is always saved to file as soon as it is acquired The default setting for the Sampling Interval is 60 seconds If you wish to change the sampling interval type in a new value in units of seconds Notice that the sampling interval is individually configurable for all available sensors STEP 6A Select the Device Configuration tab at the top of the page The table column labeled Ave Time 2 min determines the averaging time constant setting for the BE2100 sensor data The averaging time constant determines how quickly the sensor responds to change The larger the time constant the smoother the data and the slower the response The default setting is 2 minutes To set the averaging time to a new value click on the current value and select from among the choices ranging from 0 seconds to 8 minutes Note that as soon as you select a new value it is written into sensor memory The new value will persist across power cycling and will automatically be loaded into the table if you restart the program The settings are individually configurable for each BE2100 sensor STEP 6B OPTIONAL The Device Configuration Table column labeled Growth Window 8 min determines the time window over which the determination of growth rate is performed The growth rate is determined from a linear fit to the natural logarithm of the biomass vs time During expon
27. ec Mio r e B a e k Pressing the More button brings up screens with additional time constants Pressing the button beneath any of the times 72 lil will set that as the new time constant 2min is the default setting Selecting the Show key brings up a screen in which the current setting is displayed AO1 Range Pressing the button below AO1 for BE2100 base units or Range for BE2400 base units brings up the Analog Output Range menu li 5 Amal o g Out put Ria n g Set Hel p Show Bac k Set Pressing the Set button allows the user to toggle between the allowable selections for this field 0 01 0 1 1 0 10 100 1000 where 100 is the default setting Show Selecting the Show key brings up a screen displaying the current range value for 3 sec after which the screen returns to the main AO1 Range screen AO Range 2 AO2 BE2100 base units only If the button below AO is pressed the Analog Output2 Menu is displayed ii Amal o g Ou t u t 2 Me n u Ria n g je Win d w Baek Range Pressing Range button brings up the following screen Anal og Output 2 Ra n g Set Hel p Show Baek 5
28. is displayed for each sensor on the All Sensors graph is determined by the data type that was selected on the individual sensor plots Thus if the Calibrated data type was selected in the Sensor 1 graph window the data that will be displayed for Sensor in the All Sensors window will also be Calibrated The sensors that are displayed in the All Sensors graph can be controlled by using the On Off column in the Multi Plot Settings Table The relative positioning of the different sensors can be editing the X Offset Y Offset and Y Scaling values in the Multi Plot Settings Table Annotations can also be added to the All Sensors graph However these annotations are only saved in temporary memory and are not written to file In order to save annotations into permanent record they must be marked on the individual sensor graphs Terminating Data Collection Selecting STOP terminates data collection USER CALIBRATION If you wish to convert the BE2100 sensor data into reference units other than Bug Units you will need to apply a calibration Your BE2x00 software can help you to collect and then apply a custom calibration file When applying your calibration file you will no longer need to perform aliquot extraction or any other classical method in order to determine biomass or related quantities as the BE2100 sensor will now do that for you Collecting Calibration
29. left on bioreactor during sterilization cycle 1 Make sure that the last 2 digits of the calibration cup serial numbers match with that of the sensor 2 Clean the Sensor Window and or Verification Cup See Verification of Sensor Performance 3 At the end of the Sensor Check procedure update the sensor coefficients See Verification of Sensor Performance 4 The sensor will need to be returned to BugLab so that the laser can be replaced and the sensor re calibrated Biomass readings are not stable 1 Sensor position needs to be optimized 2 The sensor averaging time constant needs to be optimized 1 See Setting up for a Bioreactor Run Step 4 2 Set the sensor averaging time constant to the highest value allowed by the growth rate of the culture See step 6A of Setting Up and Configuring 92 Sensor Calibration Trouble Shooting Observation Extrapolating Cal Possible Causes The displayed biomass is outside the range of the user generated calibration Suggested Remedies This message is provided for informational purposes No action is required Below range The displayed biomass is below the range of the internal sensor calibration This message is provided for informational purposes As the biomass increases this message will automatically disappear User cal error Calibration transform is set to Log Log and negative
30. only allowed if the serial number of the currently selected BE2100 sensor matches that in the file header of the previously written data file Also all settings are read from the old data and event file and used to update the sensor configuration before the appended data acquisition commences Selecting Change will return you to the Select a filename for data storage Window STEP 3 At this point you are now actively collecting data and it will begin to appear on the graph window for the selected sensor number New data points will appear at the time interval you previously chose see Setting up and configuring step 5 so do not be alarmed if you do not immediately see new data points appearing on the graph When data collection is started the Biomass Readings table is automatically selected in the top portion of the program window Each time a new reading is taken by the sensor this table is updated with the new time stamp and biomass reading The biomass reading displayed in this table is processed according to the configuration settings if baseline correction is on then this result will be baseline corrected and if user calibration is on the biomass will be reported in user calibrated units A numerical Error Code and its interpretation are also displayed in the table An error code of 0 and the error code interpretation Normal Operation are displayed when the sensor is operating in the normal range If the sensor is operati
31. or zero valued data encountered 1 Make sure the baseline is correctly set AND 2 Wait for the biomass to increase above 0 OR 3 Switch to the linear transform method See Editing Generating and Saving a Calibration Base Unit Trouble Shooting Observation The base unit keypad is un responsive Possible Causes Base unit is in remote mode Suggested Remedies 1 Exit out of the BE2x00 Virtual Instrument software 2 If the keypad is still un responsive turn the base unit off and back on again The base unit is requesting a password but the password is lost A prior user has turned on password protection of the configuration settings See Password Locking and Unlocking Multiple sensors are plugged into the BE2400 base unit but only 1 sensor result is being displayed on base unit screen Scrolling mode is Off 1 If operating in local mode use the keypad to set the display mode to Scrolling See Introduction to the BE2400 Base Unit Functions 2 If operating in remote mode go to the Device Configuration window and check the Base Unit Display Scrolling settings Analog output is not responsive to changes in sensor readings Range setting is not optimal Make sure the AO range setting matches with the maximum anticipated biomass reading See Working with the BE2100 and BE2400 Base Units Analog Output 93 BE2x00 Software Communi
32. packet whose value indicates that the system is busy error code 8 Table 1 Commands Recognized by the BE2100 Optical Sensor Head and passed through by BE2100 Base Units BE2400 Base Units and BEIUSB adapters when connected to sensors Key PW Command Description Key Return Data Description A R W Averaging Time a Averaging Time Constant Constant B R W Baseline Value b Baseline Value K R W Start Query End k Baseline status Baseline L R W Sensor Check 1 Sensor Check Function Control Function Status M R Get data m Data output O R W On Off settings for o On Off settings for Base Corr and User Base Corr and User Cal Cal S R Sensor Serial Numbers S Sensor Serial Numbers T R W User Cal Units t User Cal Units U R W User Cal Coefficients u User Cal Coefficients V IR Sensor embedded SW v Sensor embedded SW Version Version W R W Slope Window w Slope Window Command not recognized Table 2 Commands Recognized by BE2100 Base Units BE2400 Base Units and BEIUSB adapters Key Return Data Description Command Description IR Get Base Unit 2 Base Unit embedded embedded SW version SW version R Base Unit Serial 3 Base Unit Serial Number Number 78 Table 3 Commands Recognized only by BE2100 and BE2400 Base Units Key PW Command Description Key Return
33. selected all of the data will always be stored to your selected data file Note autoscaling can also be turned on and off by clicking on the lock symbol within the scale legend You can use the cursor to read the value of a specific data point First select the cursor tool which is depicted with cross hairs and is located to the bottom left of the graph Next use the cursor positioning tool four diamond shapes located at the bottom center of the graph to move the cursor to the desired location The cursor can also be moved with the mouse by left clicking the mouse over the cursor and dragging the cursor to a new location whiling keeping the mouse button depressed The X and Y values of the data point where the cursor is located are indicated within the cursor box at the bottom right of the graph By right mouse clicking over the cursor box several other options can be accessed The cursor can be centered within the screen by selecting the Bring to Center option Additional cursors can also be created or deleted You can change the displayed data type by selecting the switch at the upper left of the graph to Raw Baseline Corr Calibrated Growth Rate or Error Code Note that the unless otherwise selected the displayed data type will be set according to the Device Configuration table if User Cal is On then the data type will be Calibrated if User Cal is Off and Baseline Corr is On then the data type will be B
34. the BE2x00 Virtual Instrument software and wait for the list of detected BE2x00 devices to finish updating gt o END 95
35. the front panel of the Base Unit vs time in hours The growth rate time window variable determines the data set to be used for the slope calculation The range settings for AO1 and AO2 and the time window for AO2 can be set by 3 different methods 1 the keypad interface on a BE2100 or BE2400 Base Unit 2 the Virtual Instrument Software and 3 by sending serial commands to the Base Unit Further description of the keypad interface can be found in Appendix II of this manual Part II of this manual describes how the Virtual Instrument Software can be used to 29 change the settings Appendix III of this manual describes the serial commands R and W that can be used to programmatically set these parameters Note 1 The screw terminal for connecting the analog outputs is removable It is supplied separately from the base unit and can be inserted by simply pushing it into the slot provided in the rear on the Base Unit This feature makes it easier to connect wires to the screw terminals Note 2 The maximum resistance that the Base Unit can encounter to drive a full 20 mA is 500Q By connecting a 500Q resistor across the and terminals the analog output can be converted from a nominal 4 20 mA current source to a nominal 2 10V voltage source Two 500Q resistors are provided as accessories for this purpose Note 3 The 4 and 20 mA current levels are described above as nominal values because the actual
36. then the Next button STEP 5 A summary of the software components that are about to be installed is next displayed Click on the Next button to begin installation Installation may take several minutes STEP 6 If the software has successfully been installed you should see an Jnstallation Complete window Clicking on the Next button STEP 7 The driver for the USB virtual communication port device in the OD Scanner instrument is next installed Ifa windows pop up message asks whether you want to allow changes to your computer select Yes A window entitled FTDI CDM drivers should open Select Extract Once extraction is complete the Device Driver Installation Wizard should automatically start Select Next and wait for a confirmation that the driver was successfully installed Select Finish to exit out of the Installation Wizard 38 Setting Up and Configuring STEP 1 Configure the power settings on your computer so that sleep mode is disabled Step i Select Start Control Panel in Windows 8 from the desktop simultaneously press the Windows and the C key and select Settings and then Control Panel Step ii Select Hardware and Sound Power Options Edit Plan Settings Step iii Choose Never for the Sleep setting Step iv Press the Save Changes button and then exit out of the Control Panel window Note Serial communication is interrupted when a windows c
37. unit press the More button then select the sensor number for which you want to set the baseline At this point 27 the bottom line of the display should read Base Cal More or Exit Note 1 If the Virtual Instrument software is currently communicating with the BE2100 base unit as indicated by Remote Operation on the top line of the display keypad access is locked out You will either need to suspend communication or perform the baseline correction from within the Virtual Instrument software section II E 2 Note 2 If you are not the 1 user of the instrument and a prior user has turned on password protection before you can reach the above configuration menu you will be prompted to enter a password You may wish to consult the prior user to obtain the password Alternatively if the password has been lost or forgotten the password may be reset via the user interface software If this is the case skip this section for now and come back to it later after you have installed the user interface software and reset the keypad password STEP 1 Press the Base button STEP 2 Select the Start button and wait at least a few seconds before selecting the Stop button Between the time that the Start and Stop buttons are pressed the sensor readings will be averaged The averaged values will be used to determine the new baseline setting After the Stop button has been pressed the ne
38. values will vary slightly from instrument to instrument For best accuracy when working with the analog outputs it is recommended to measure the actual currents or voltages produced when the signal is at 0 and when it is at or above the maximum determined by the range setting An example calibration procedure is provided in Appendix IV of this manual Note 4 The digital to analog converters DAC in the base unit provide 12 bit precision meaning that the minimum step size is 4 WA In order to ensure best performance it is important to match the Range setting to the maximum anticipated biomass reading Keypad Lockout Many functions of the BE2x00 instrument may be accessed both by keypad interface on a Base Unit and programmatically through serial interface commands In order to prevent conflict between these different methods of accessing the same functions it is helpful to establish a prioritization between them When the BE2x00 is being controlled remotely through the Virtual Instrument software access to the keypad is restricted For users writing their own interface code it is recommended that access be similarly restricted using the J command described in Appendix IIT When the Virtual Instrument software is running the message Remote Operation will be displayed on the top line of the BE2100 or BE2400 Base Unit display In this state the keypad configuration menus cannot be accessed The Base Unit always starts up
39. window as is provided in the BE2x00 Virtual Instrument program See the section Editing Generating and Saving a Calibration for a full description of how to operate the controls in this window Note To view calibrations without having to first open a data set use the Cal Window button provided in the User Calibration tab in the top portion of the program window Within this 55 window you can modify and save new calibrations without applying these changes to a data file STEP 4 Ifthe Calibration control if set to Off turn it On Press the OK button to put your changes into effect If you want to save your changes to file press the Resave button select a new filename and then press the OK button 56 End User License Agreement IMPORTANT READ CAREFULLY This End User License Agreement EULA is a legal agreement between you either individually or a single entity and BugLab LLC BugLab By installing copying or otherwise using the BE2x00 software you agree to be bound by the terms of this EULA If you do not agree to the terms of this EULA Buglab is unwilling to license the BE21x00 software to you In such an event you may not use the BE2x00 software and should contact BugLab for instructions on the return of the product for a full refund Software Product License The BE2x00 software is licensed not sold 1 GRANT OF LICENSE This EULA grants you the following
40. 1 will read Extrapolating Cal and line 2 will continue to report the quantitative result 2 Error conditions a b If ambient light is high enough to prevent a valid measurement then line 1 will read Signal Saturated and line 2 will be blank If the sensor is disconnected from the base unit then line 1 will read Sensor Disconnected and line 2 will be blank If the sensor measures signals that are below the internal calibration range then line 1 will read Below Range If the sensor measures signals that are above the internal calibration range then line 1 will read Above Range If there is an error converting the data into BugUnits then line 1 will read Data Error and line 2 will be blank Additional errors and warnings are shown in the table on the following page 67 Error and Warning Codes Code Typeof Useage Condition Notes Event 10 Error C Check Sum Sent check sum does not match computed Error check sum 9 Error C Value out of This is a serial communication error where range the data packet following a command contains values outside of the allowed range 8 Error C Network busy This general error code applies when any of the 3 communicators sensor monitor or PC is busy with another task when any other of the communicators is attempting to talk with it 7 Error G User Attempted to fit negative or zero value
41. 100 sensor is provided with a pair of low and high calibration cups with unique serial numbers Due to small variations between the calibration cups the same calibration cups should always be used with the sensor with which they were provided STEP 1 Check that the front face of the sensor is clean and free of fingerprints The sensor face may be cleaned with commercial window cleaner ethanol or isopropanol using lint free tissue DO NOT USE ACETONE as it may cause irreparable damage to components used in the sensor Likewise check and if necessary clean the surface of the Low and High calibration cups that come in contact with the front face of the sensor If you have more than one BE2100 sensor note that the calibration cups provided for testing sensor performance are not identical and should be matched with the corresponding sensor The last 2 digits of the serial numbers printed on the sensor and low and high calibration cups should all match each other STEP 2 Remove all of the magnetic adapter sleeves from the sensor Place the Low calibration cup over the front of the sensor Make sure that the arrow marked on the calibration cup is pointing towards the arrow marked on the sensor Connect the buckle components attached to opposite ends of the strap Cinch the sensor against the calibration cup by pulling on the free end of the strap that protrudes from the buckle Next close the latch on the back of the
42. Data J R W Base Unit keypad j Base Unit keypad lockout state lockout state Q R W Base Unit password q Base Unit password set reset unlock status R R W Range for Analog Out r Range for Analog Out Table 4 Commands Recognized only by BE2400 Base Units Key PW Command Description Key Return Data Description R W Sensor port switch state 5 Active sensor port Notes for Tables 1 4 1 The symbols in the PW column indicate the type of access that is available Read R Write W or both R W 6 Command Descriptions 1 A Ox41 Averaging Time Constant The time constant in seconds used in the sensor for averaging the raw detector data The value is represented as an unsigned integer16 Allowed values 0 30 60 120 240 480 Any non allowed value entered is ignored Default value is 120 Key Time Window 1 Byte 2 Bytes A 0x41 Sending an empty packet A command will return the a message displaying the current setting Key Time Window 1 Byte 2 Bytes a Ox61 2 B 0x42 Baseline Value The offset value applied to the sensor ouput in order to compute the Baseline Corrected sensor output Values expressed as 32 bit floating point number Default is 0 0 79 Key Baseline to Set 1 Byte 4 Bytes B 0x42 LSB MSB Sending an emp
43. E2400 Base Unit has 4 analog current outputs one for each sensor These analog outputs provide the same functionality as AO1 on the BE2100 Base Unit AO reflects whatever corrections are applied to the data being displayed on the base unit So if baseline correction is on then AO1 will also be baseline corrected Similarly if user calibration is on then AO1 will be in user calibrated units The growth rate calculation used to determine AO2 is independent of the units in which biomass is expressed However the baseline setting will affect this calculation so particularly at low biomass it is critical that the baseline is set so that a baseline corrected sensor reading of zero corresponds to zero biomass A signal of zero will always correspond to a nominal value of 4 mA on AO1 while a growth rate of zero will always correspond to a nominal value of 4 mA on AO2 The minimum signal that corresponds to a nominal value of 20 mA on analog output 1 will be determined by the analog output range variables see key pad communication Appendix II section 1 v for further details Any signal higher than this minimum signal level will cause AO1 to output a nominal value of 20 mA The minimum growth rate that corresponds to a nominal value of 20 mA on AO2 is determined by the range setting for AO2 The rate of change reflected in AO2 is determined by a linear least squares fit to the natural logarithm of the data type currently being displayed on
44. Lab provides no warranties whatsoever used in connection with any BugLab device express or implied Neither does it guarantee software compatibility with any off the shelf software package or any software program that has not been written by BugLab Intended use of this system must be followed within the guidelines of this manual In no event will BugLab be liable for any damages caused in whole or in part by any customer or for any economic loss physical injury lost revenue lost profits lost savings or other indirect incidental special or consequential damages incurred by any person even if BugLab has been advised of the possibility of such damages or claims The optical designs and circuit board designs in the BE2x00 products are proprietary to BugLab The user may not copy any of the designs either in whole or in part without written permission from BugLab Windows is a registered trademark of Microsoft Corporation The BE2x00 software is written in the LabVIEW development environment Copyright 2015 National Instruments Corporation All Rights Reserved Copyright BugLab LLC 2015 All Rights Reserved Cautions Viewing the laser output with certain optical instruments for example eye loupes magnifiers and microscopes may pose an eye hazard gt The low voltage BE2100 sensor is water resistant but it is not water proof Keep the sensor the Base Unit the cables and power supply clean and dry Do no
45. TM bug lab OPERATOR S MANUAL BE2x00 Noninvasive Biomass Monitor User manual for the following BugLab products BE2100 sensor BE2100 base unit single sensor BE2400 base unit 4 sensor multiplexing BEIUSB adapter Includes Instructions for BE2x00 Virtual Instrument and Data Viewing Software BugLab LLC www buglab com info buglab com Last updated February 17 2015 Notice This publication and its contents are proprietary to BugLab LLC BugLab and are intended solely for the contractual use of BugLab customers While reasonable efforts have been made to assure the accuracy of this manual BugLab shall not be liable for errors contained herein nor for incidental or consequential damage in connection with the furnishing performance or use of this material BugLab reserves the right to revise this manual and make changes from time to time without obligation by BugLab to notify any person of such revisions or changes BugLab does not assume any liability arising out of the application or use of any products circuits or software described herein Neither does it convey a license under its patent rights nor the patent rights of others This publication and its contents may not be reproduced copied transmitted or distributed in any form or by any means radio electronic mechanical photocopying scanning facsimile or otherwise or for any other purpose without the prior permission of BugLab Bug
46. al Instrument and Data Viewing software are provided in Section 2 of this manual 7 Optional Connections Required Connection BE USB offers USB connectivity only GETTING STARTED Unpacking the Instrument The BE2100 sensor consists of Optical Sensor Head with cable 2 m Sensor Adapter Sleeves 10 15 and 20mm Calibration Cups Low and High Additional components provided with a BE2100 base unit BE2100 Base Unit Power Adapter RS 232 and USB cables 2 m Analog output terminal block 4 position Resistors 500 Q for optionally converting 4 20 mA analog outputs into 2 10 V outputs BE2x00 Virtual Instrument and Data Viewing Software Additional components provided with a BE2400 base unit BE2400 Base Unit Power Adapter RS 232 and USB cables 2 m Analog output terminal block 8 position Resistors 500 Q for optionally converting 4 20 mA analog outputs into 2 10 V outputs BE2x00 Virtual Instrument and Data Viewing Software Additional components provided with a BEIUSB adapter BEIUSB adapter USB cable 2 m BE2x00 Virtual Instrument and Data Viewing Software 8 Optional accessories Extended 7 5 m RS 232 cable Extended 5 m USB cable Screw mounting Sensor Attachment Kit Sensor Adapter Sleeve 5 mm Sensor cable extender 4 m Sensor strap extender 2 m Note Unpack and inspect all of the components to assure that they have not been damaged in shipping
47. alic text is used to indicate window names 3 Bold italic text is used to indicate sections of this manual 4 The sign is used to indicate sub levels of menu commands For example Start Settings Control Panel Add Remove Programs means select the 4 Start menu then select the Settings sub menu further select the Control Panel sub menu and then finally select the Add Remove Programs sub menu 37 Software Installation Note Before plugging a BE2x00 device into your computer make sure you have installed the software first STEP 1 Insert the BE2x00 software Compact Disk CD into your computer STEP 2 If a Windows AutoPlay message pops up under Install or run program from your media choose Run setup exe If the installation does not start automatically browse the CD for the setup exe file and double click on it If Windows asks Do you want to allow the following program setup exe to make changes to this computer click on the Yes button STEP 3 You will be prompted to choose destination directories for both the BE2x00 BugLab software and the LabView National Instruments run time engine used by the BE2x00 software Click on the browse buttons if you want to install these programs somewhere other than the default directories shown Otherwise click on the Next button STEP 4 Review the software license agreements If you agree then click on the I accept buttons and
48. aseline Corr if both User Cal and Base Corr are Off then the data type will be Raw By selecting the Growth Rate data type you can observe a real time estimate of the exponential growth rate of your organism in units of inverse hours By 46 selecting the Error Code data type you can quickly identify whether any data points were collected under conditions of error negative error codes warning positive error codes or normal operation error code 0 Note In all of the display views data points that were collected under normal conditions are displayed in light green Data points collected under warning or error conditions are depicted in yellow or red respectively 4 By right clicking on the data markers within the plot legend to the upper right of the graph you can change many aspects of the graph including the plot style marker color and marker symbol Recording Events during Data Collection A helpful tool provided in the data acquisition graph windows is annotation or event marking As you alter conditions during a bioreactor run you can easily note them and they will be automatically time stamped Note Data acquisition must be started before the create annotation feature becomes active in the individual sensor graphs STEP 1 To record an event simply right click on the area of the plot where you wish to record an annotation and select Create Annotation Several pre defined event types ar
49. aseline Adjustment The Data Viewer program also provides the capability to retrospectively adjust the baseline that is applied to the data This feature is useful when you want to apply the same baseline setting across the entire file If the baseline was changed one or more times during data acquisition with the Virtual Instrument program the changes were applied prospectively In such cases different segments of data have different baseline settings The retrospective baseline adjustment feature allows you to equalize the baseline setting across all data segments STEP 1 From within one of the File tabs press the Open button select the file you want to work on and then press OK 53 STEP 2 From within the same File tab press the Adjust Base button The Set Baseline Retrospectively vi window will open STEP 3 Set the Baseline Method control to one of the three settings Set manually If you select this option you should type the new baseline value directly into the Baseline Value control Retrieve from config table If you select this option the value that is in the Device Configuration table in the top half of the program window is written into the Baseline Value control Compute from graph If you select this option the baseline value is determined by averaging all points between the Baseline Start and Baseline End annotations If
50. at the front faces of the sensor and calibration cup are clean and dry and that the calibration cup is securely attached to sensor see step 2 above It will be necessary to repeat the low calibration cup step 11 above measurement before proceeding again with the high calibration cup measurement If the readings were stable a Pass Fail assessment of sensor performance will be made This Pass Fail assessment is based on a comparison of the present measurements to prior measurements made on the calibration cups during manufacture STEP 4 Whether Pass or Fail is indicated at the end of the Calibration Cup measurements you will be given the option of updating the sensor calibration coefficients based on the measurements just completed If the update or Set New Coeff in the Virtual Instrument software option is selected new sensor coefficients will be written into sensor memory The new coefficients are determined by linearizing the newly completed calibration cup measurements to the original calibration cup measurements collected at the time of sensor manufacture These new sensor coefficients will persist even across power cycling of the instrument However it is important to realize that selecting the Reset function either via the keypad or the Virtual Instrument software will reset sensor coefficients to their original factory settings SETTING UP ON A BIOREACTOR Attaching the Sensor Head STEP 1 Posi
51. ation button at the bottom left of the pop up window STEP 5 You have now defined the start and end points for baseline determination so you are ready to set the baseline Right click the mouse button anywhere over the graph From the drop down list that appears select Create Annotation gt gt Baseline Set STEP 6 The Set Baseline Interactively Window now appears When this window is brought up the current baseline value is automatically read from the sensor and is displayed in the Baseline Value box To define a new baseline based on the start and end points you have just selected choose Compute from graph in the box labeled Baseline Method The newly computed value is displayed in the Baseline Value box Alternatively if you wished to set the baseline manually you could have selected Set manually as the Baseline Method Then you could manually enter a new value into the Baseline Value box Note that when the Set manually option is selected the current sensor reading is written into the Baseline Value window but you may edit this value if desired Select the OK button at the bottom left of the pop up window You have now set a new baseline value The time at which the new baseline value was set is recorded and displayed in the text box in the main window In addition the annotation is numbered and marked on the graph Notice that the baseline is applied only pros
52. by hand until it reaches a stop Do not use tools to further tighten the connector STEP 2 Connect the power adapter cable to the Base Unit or the USB cable to the BEIUSB adapter STEP 3 Plug the power adapter into a power source and switch the unit on or plug the USB cable coming from the BEIUSB adapter into your computer and the sensor will automatically be powered on Introduction to the BE2100 Base Unit Functions The keypad interface on BE2100 and BE2400 base units provides access to many functions a full description of which can be found in Appendix II Because the BE2400 base unit connects to up to 4 sensors the keypad interface is a little different than that of the BE2100 Skip to the next section Introduction to the BE2400 Base Unit Functions if you are using a BE2400 base unit All of the functions available via the keypad interface are also available through the Virtual Instrument Software If you are using a BEIUSB adapter or plan to operate the instrument solely through the Virtual Instrument software you can skip this section However if you wish to operate a BE2100 base unit in stand alone mode without a computer the following tutorial may be helpful This tutorial will give you familiarity with the basic functioning of the keypad interface by leading you through the steps required to modify one parameter the sensor response time 10 STEP 1 Press any of the 4 arrowhead keys on the front of the Base U
53. calibration data yet the Calibration value will be 1 Add the off line reference values to the Calibration column Make sure that the Event corresponds to the correct sample The data displayed on the graph will be updated as soon as you enter it into the table STEP 3 If necessary adjust the Raw Sensor and Calibration measurement offsets by using the Adjust Baseline Fixed Intercept and Intercept controls The value in the Baseline column of the calibration table is subtracted from the Raw Sensor readings before correlating them to the Calibration values By pressing the Adjust Baseline button you can simultaneously change the Baseline setting for all measurements in the Calibration Table Individual Baseline values can be adjusted by directly editing the values in the Baseline column of the Calibration Table When the Fixed Intercept control is checked the linear fit is forced to intersect with the y Intercept at an OD Scanner reading of zero It is recommended that you keep the Fixed Intercept control checked unless you have entered Reference values that span both the low and high range of the biomass units into which you are calibrating When the Intercept is fixed generally it is recommended to keep it fixed at zero unless the reference method has a built in offset that you haven t already accounted for e g if you measured OD in a complex medium and didn t zero the spectrophotometer using the media alone then you shoul
54. cation Trouble Shooting Observation Possible Causes Suggested Remedies BE2100 sensor not 1 Sensor USB or RS 232 1 See Setting up and detected by BE2x00 cable not connected Configuration Virtual Instrument 2 Base Unit is in local 2 Exit out of the configuration software mode menus 3 RS 232 and USB 3 If communicating via RS 232 communications are in make sure the USB cable is conflict unplugged from the base unit If 4 USB driver was not communicating with a BE2100 correctly installed base unit via USB make sure the 5 BE2x00 device not comm port setting is USB See recognized by a USB hub Setting Up and Configuring 4 See USB Driver Trouble Shooting at the end of this table 5 Re boot your computer Computer 1 The computer went into 1 See Step 1 of Setting Up and communication with the sleep mode Configuring BE2x00 device was 2 Power to the BE2x00 2 Consider connecting the interrupted device was interrupted BE2x00 device to a power source 3 Connecting through a that can provide uninterrupted USB hub device power e g battery back up 3 If possible connect your BE2x00 device directly to a USB or RS 232 port on your computer rather than connecting through a USB hub which we have found to be less reliable USB Driver Trouble Shooting If you are connecting the BE2x00 instrument to a computer via USB but the instrument is not recognized by the User Interface software
55. ck The Back button allows the user to step back in the menu b Calibration Mode Cal If the button below Cal is pressed the Calibration mode is entered Seer eee ere pierce Eee tess eae ese ete AJ A AJ 4 The buttons in this mode will have the same functions as in the baseline mode 70 C Sensor Check Chk Ifthe button below the Chk is pressed the Sensor Check mode will be entered Sems or Che c k Mio de Riu in He l p B a e k i Back Ifthe Back button is pressed here and in all of the following screens in Sensor Check mode unless otherwise noted the previous main configuration menu will be displayed again ii Run Pressing the button under Run begins the following programming sequence At t a c h Lo w Cal Cup Re a d y Hell p B a ce kk iii Ready Pressing the Ready button leads to 25 measurements 1 sec each being made The mean and standard deviation are computed During the measurement the base unit displays the message Meas uri neh i xX o 25 where X is the current measurement number 1 24 and is updated following each measurement except for the 25 iv Once the 25 measurements are complete the
56. comparison of how much light is extinguished within a sample when the chromophore is present at different concentration levels e g zero and a known concentration By contrast in a scattering measurement light is deviated from its path rather than being extinguished As a result the measured amount of scattering will be dependent on the area and angle of scattered light that is captured by the detection system Since the detector size and geometric arrangement is not standardized between different commercial spectrophotometers the Optical Density determined for biomass samples can vary significantly e g 50 or more variation between different spectrophotometer models For this reason if you would like the BE2100 sensor to report results in OD units it will be necessary to calibrate to the specific spectrophotometer that is used for the off line OD measurement A simple step by step guide for generating a custom calibration for the BE2100 Sensor is described below User Calibration Section II G The relationship between back scattered light intensity as measured by the BE2100 and biomass such as dry cell weight is weakly dependent on the size of the scattering particles For this reason it is recommended that separate calibrations be used for organisms with grossly different cell sizes such as Escherichia coli typical cell diameter 0 5 1 um and Saccharomyces cerevisiae typical cell diameter 5 10 um For mono disperse cell cult
57. d data Calibration in log log space Error 6 Error G User Insufficient number of calibration points to Calibration compute calibration coefficients Error 5 Error D G Signal Ambient light is so high that it is preventing Saturated measurement 4 Error D G Sensor Sensor not plugged into monitor Disconnected 3 Error D G Below Range Signal is below the internal cal range 2 Error D G Above Range Signal is above the internal cal range 1 Error D G Data Error Error converting the individual detector data into BugUnits 0 None D G Normal Operation 1 Warning D G High Ambient Ambient light is high but a measurement can Light still be made 2 Warning D G No The calibration switch is on but no Calibration calibration data is available 3 Warning D G Extrapolating The measurement is outside of the user Cal calibration range Notes for Table 1 1 D base unit display G graphical user interface C serial communication error 68 v 4 key communication 1 The variables and functions in the order in which they will appear on the menu are a k Scroll Lock BE2400 base unit only In scroll mode the results for attached sensors are sequentially displayed for 5 seconds each In locked mode only the results for the selected sensor are displayed Baseline Base The user will have the ability to set a baseline and turn baseline correction On or Off In the On state the most rec
58. d enter the OD of the medium alone as the Intercept value STEP 4 Enter the type of reference data into the Calibration Units box at the top left of the graph It is recommended that you include means of identifying both the organism and the reference method e g e coli g L But be aware that if you type in a name that is longer than 10 characters including spaces it will automatically be truncated to 10 characters STEP 5 Choose the data transform method Generally the Linear Linear is recommended If your samples were collected 51 at logarithmic intervals the Log Log transform may work best However be aware that an error will be reported if the baseline corrected Bug Units value is less than or equal to zero STEP 6 Choose the Polynomial order that will be used to fit the data We recommend using the lowest polynomial order that adequately fits the data Due to the high linearity of the BE2100 sensor response to biomass a linear fit polynomial order 1 is generally recommended The root mean squared RMS error and linear correlation coefficient R for the fit are shown below the graph RMS Error indicates the root mean squared difference between the linear fit and the actual data so the smaller the RMSE the better the fit R values can range between 0 and 1 with 1 indicating perfect linear correlation The polynomial coefficients Poly Coeff resulting from the fit are also displayed bel
59. d ignored by the sensor Sending an empty L command packet will return the I message along with a data packet The first Byte of the packet holds a status message The meaning of the status messages are shown below 1 Values Meaning 0x00 No data Ox11 Busy with Low cal cup measurement 0x12 Busy with High cal cup measurement 0x13 Busy with setting new Variable Sensor Coefficients 0x21 Low cal cup measurement was unstable 0x22 High cal cup measurement was unstable 0x31 Low cal cup measurement was stable but Combined Result failed 0x32 High cal cup measurement was stable but Combined Result failed 0x41 Low cal cup measurement was stable and Combined Result passed 0x42 High cal cup measurement was stable and Combined Result passed 0x53 Error new Variable Sensor Coefficients could not be set 0x63 New Variable Sensor Coefficients were successfully set If the Sensor Check routine has just been started either through the interactive keypad or the user interface software and no calibration cup measurements have been initiated yet sending an empty L command to the sensor will result in an I value of 0x00 No data being returned If a calibration cup measurement is in progress when an empty L command is sent to the sensor a Busy 0x11 or 0x12 I value will be returned to the sender When a calibration cup measurement has been completed sending an empty L command to the sen
60. different combination of adapters or none In such cases the adapters should be selected so that the sensor front face is in intimate contact with the port window while the adapter s provides a stable mounting surface on the window flange Note that alternate and custom screw mounting options as opposed to the strap mounting method described here are also available for some bioreactor types Inquire at BugLab info buglab com for more details Pass the black mounting straps around the bioreactor If the bioreactor has external metal rods thread the straps under them if possible so that the straps are only in contact with the body of the bioreactor Avoid twists in the straps the strap should lie flat around the perimeter of the bioreactor Close the loop around the bioreactor by connecting the two buckle components attached to opposite ends of the strap note if the strap is not long enough strap extenders are available Cinch the sensor against the bioreactor by pulling on the free end of the strap that protrudes from the buckle Next close the latch on the back of the sensor this step should provide the final pressure needed to firmly seat the sensor against the bioreactor Do not use pliers or any gripping tools that would exert excessive force The goal is to snug the sensor down on its black flexible gasket once the latch is closed so that the sensor is as close to the surface of the bioreactor as possible If by pushing on t
61. e available including Inoculation Calibration Sample Removal Sparge Rate Change Agitation Rate Change Foam Breaker Rate Change pH Change Temperature Change Nutrient Addition Induction Harvesting Baseline Start Baseline End Baseline Set Delete All Annotations 47 You also can enter a User Defined event for non standard events STEP 2 Select one of the event labels from the list STEP 3 In the Add Annotation window various options can be set and recorded depending on what event is selected Additional comments can be added within the text box labeled Edit the annotation text below if desired A Value and Units associated with the event can also be entered For example if the agitation rate was set to 500 rpm 500 could be entered into the Value box and rpm could be entered into the Units box The positioning of the annotation on the graph is determined by the Positioning Method selector box The annotation can be added to the end of the dataset corresponding to the moment in time when Add Annotation was initiated or it can be added at a manually selected point in time time can be specified under the Graph Position label or it can be added at the cursor position where the mouse was clicked STEP 4 When all parameters have been satisfactorily edited click on the Add Annotation button The annotation is numbered and marked on the graph as wel
62. e overall organization and appearance of the program is much like that of the BE2x00 Virtual Instrument Previously acquired files are opened using the Open button from within the File tabs located in the bottom half of the program window Files can be re saved using the Resave button but a new filename must first be selected A suggested filename is automatically generated that consists of the original filename appended with the date and time at which it was resaved Resaving with the original filename is disabled in order to protect against accidental overwriting of data This is particularly important when the Data Viewer program is used to open a file into which data is still actively being acquired by the Virtual Instrument program The data that is displayed in the Data Viewer program is a copy of the data that was present at the time the file was opened the viewed data file is not automatically updated as new data points are collected As with the Virtual Instrument program the last tab in the files tabs contains a graph in which data from multiple files can be overlaid The configuration tabs in the top half of the Data Viewer screen are also similar to those within the Virtual Instrument program However in the Data Viewer the configuration settings cannot be modified read only These configuration settings are as read from the header of the data files that have been opened Retrospective B
63. e sent as a Command Response pair The response key is the lower case complement of the command key except in the case where the command is not recognized If a command is not recognized by the receiver then a special ASCII character hex 0x21 is returned to the sender with a 1byte data packet Empty command packets will be used to prompt for the current settings to be returned Non empty command packets are used to set the parameters for the specified field value provided the packet and parameters for the field meet specifications If the data in a non empty command packet is successfully received the response will include a duplication of the data in the command packet If the data in a non empty command packet is out of the allowed range of values then the response will contain a 1 byte data packet with a value 9 that indicates that the data was Out of Range If the computed check sum does not match the sent check sum then the response will contain a 1 byte data packet with a value 10 that indicates that there was Check Sum Error If data packet is both out range and contains a check sum error the response data packet will indicate Check Sum Error In either case other than responding with the error message no action will be taken by the receiver in response to the command sent Any 32 bit floating point value is represented in IEEE 754 format unless otherwise specified 3 Data Protocol Packe
64. eatures Access to individual sensor and base unit settings through interactive Keypad Password control optional of keypad access Lighted display with variable brightness control Digital USB and RS 232 and analog 4 20mA or 2 10V outputs biomass and growth rate User Interface Software Features Real time graphical and numerical display for the sensor Event marking on graph both pre defined and user defined Baseline setting and subtraction User calibration of sensor output to any reference units Access to all sensor and base unit settings User Interface Software Requirements Windows XP Vista 7 8 82 or 64 bit Operating System Minimum of 256 MB of RAM Minimum 200 MB free hard disk space 1024 by 768 resolution or higher video adapter Available USB or 9 pin RS 232 COM port Microsoft compatible mouse CD reader required only at time of software installation F i Feed a _ 147mm 132 mm 4 J Core 1 8mm tes CE cy YY we 61 BE2100 Base Unit Specifications Base Unit Electrical DC Power In Both US and International plug adapters available 9V 1A Certifications CE marked Tested for compliance to EMC standards EN55011 and EN61000 and safety standard EN 61010 Sensor Input One BE2100 sensor Connector BE2100 se
65. ed within 15 seconds the display will revert to the Password Protect screen Otherwise you will be prompted to re enter the new 6 digit password If the password is successfully re entered the screen will display the message New password accepted Reset Pressing the Reset button bring up the following menu Res e t al il settings Rje s e t H je p Baek i Reset Pressing the Reset button brings up the following confirmation screen Clo nf i r m ries et Ye fs N jo Selecting Yes resets all variables to their factory default values including the sensor gains and offsets Selecting No returns the screen to the Reset All Settings window without changing any settings 75 Appendix III Serial Protocol Specifications 1 Scope Remote Connection to the BE2x00 Instrument via USB or RS 232 Applies to communication between the BE2100 Sensor and BE2100 Base Unit BE2400 Base Unit or BEIUSB adapter their communication with each other as well as communication protocol with a host PC 2 General Specifications 1 Serial data is transmitted over either a RS 232 serial port or over USB Serial data is transmitted at 19200 baud rate No Parity 8 data bits 1 stop bit Data transmission is non streaming only All data will b
66. eger 2 Raw Sensor Measurement Result in Bug Units 32 bit floating point 3 Baseline Corrected Measurement Result 32 bit floating point 4 User Calibrated Result 32 bit floating point 5 Growth Rate Constant 1 hours 32 bit floating point 6 User Calibration Units up to 11 ASCII bytes Key Error Raw Result Base Corr User Cal Growth Rate User Code Result Result 1 hrs Cal Units 1 Byte 1 4 Bytes 4 Bytes 4 Bytes 4 Bytes up toll Byte ASCII Bytes mM LSB MSB LSB MSB LSB MSB LSB MSB 0x6D Note the total length of the data packet is variable due to the variable length of the User Cal Units string The User Cal Units string includes a terminating null character as its last byte Therefore the maximum allowed number of non null characters in the string is 10 7 O Ox4F On Off Setting for Baseline Corr and User Cal The O command is used to turn On or Off Baseline Correction and User Calibration Values are represented as 8 bit unsigned integers Allowed values are 0 Off and 1 On with the default value being 0 Off for both functions The first byte sets the On Off state for Baseline Correction The second byte sets the On Off state for User Calibration Sending the highest value for a byte hex FF will preserve the current setting in the byte Values other than 0 1 and F will be ig
67. en properly set If the settings are not correct follow the steps in the Calibration section of this manual Once the calibration coefficients and units are properly set return to the Device Configuration table and set User Cal to On Initiating Data Collection STEP 1 Select the Sensor Tab lower part of the window corresponding to the sensor number for which you want to begin data collection Press the Start button STEP 2 You will be prompted to enter a file name for your data A default filename is automatically created which consists of the year month day hour minute and second at which you pressed the Start button Choose the directory where you wish to store the data and then modify the file name as desired and press OK The bug extension is added automatically Data will automatically be 42 saved to this file as soon as it is transmitted from the BE2x00 device If you selected a file name that already exists you will have the options of choosing another file name overwriting the existing file or appending to the existing file Note that selecting Overwrite will result in the old data file s being deleted so choose this option carefully Selecting Append allows you to continue a previously aborted experiment If you choose this option the old data will be loaded and the new data will be added to it The time expired between the earlier and the current experiment will be automatically accounted for Appending is
68. ential growth the slope of 41 this fit corresponds to the growth rate of the organism provided in units of inverse hours The Growth Window setting determines the time window over which the fit is performed If you plan to use this feature and want to change the Growth Window setting click on the current setting and select from among the allowed settings ranging from 1 to 32 minutes STEP 6C OPTIONAL The Device Configuration Table columns labeled Base Corr Off and Base Val 0 0 determine respectively whether baseline correction is applied to the data and the value of this correction Under normal operation it is recommended that the baseline correction be determined after initiating data collection see the section Setting the Baseline STEP 6D OPTIONAL The Device Configuration Table columns labeled User Cal Off determines whether a custom user calibration is used to transform the sensor data into units other than standard Bug Units Step by step procedures for generating a new calibration are described later in the Calibration section of this manual If you have not already generated a custom calibration set User Cal to Off if it is currently On If you have already generated a custom calibration and wish to now apply it to the new data you are about to collect select the Device Calibration tab near the top of the screen and check that the calibration coefficients and units have be
69. ently set baseline is subtracted from the Bug Units In the Off state the baseline is ignored The default baseline setting is Off Calibration Switch Cal The user will have the ability to view results in either calibrated or un calibrated mode The default Calibration Switch setting is Off Sensor Check Chk The user will be able to verify that the sensor is working correctly by placing standard calibration cups onto the sensor and if necessary change the sensor calibration coefficients offsets and gains Averaging time constant Ave Determines the time response of the sensor to changing signals The default averaging time constant is 2 minutes Range for Analog Output 1 AO1 on BE2100 Rnge on BE2400 Determines the signal level corresponding to the maximum current 20 mA on analog output 1 The default analog output 1 range is 100 Range and Time Window for Analog Output 2 AO2 BE2100 base units only The Range setting determines the rate of change per hour corresponding to the maximum current 20 mA on analog output 2 The default analog output 2 range is 100 The Wndw setting determines the time window over which the growth rate is computed for analog output 2 The default setting is 8 min Comm port switch Comm BE2100 base units only Determines whether the RS 232 or USB port is active on the monitor The default setting is
70. enus and return you to the normal display window Note that communication via the USB or RS 232 ports is disabled while the configuration menu is active so make sure to exit the configuration windows before trying to run the BE2100 User Interface software Now that you have a basic familiarity with the operation of the keypad interface you may want to experiment with other features Alternatively a detailed step by step description of the keypad features is available in Appendix II Once you have selected settings that are best for your application you may wish to prevent accidental changes to these settings by turning on the password protection feature Verification of Sensor Performance Before using the BE2100 sensor to measure biomass in a liquid culture you may wish to verify that it is performing as expected This step is recommended when you are unpacking and using the instrument for the first time but may also be used to periodically check sensor functionality In addition to providing a verification of performance the option of recalibrating the sensor is also provided The following procedure describes how to run the Sensor Check function via the keypad interface or the Virtual Instrument Software If you plan to operate the instrument solely via the Virtual Instrument Software you may wish to return this section later after you have installed the software described in Part II of this manual Note that each BE2
71. ers associated with the 4 20 mA analog outputs available on the rear of the Base Unit AO1 provides a signal that is proportional to the signal displayed on the front of the Base Unit when not in the Configuration menus AO2 provides a signal that is proportional growth rate in I hours of the AO1 signal The correspondence 13 between the analog outputs and the signal values is determined by the range settings available in these sub menus The time window over which AO2 is computed can also be selected within these sub menus STEP 13 Press the arrowhead beneath the word More Pressing the arrowhead beneath Comm and Disp would lead you into sub menus that would allow you to respectively change the communication settings and display brightness STEP 14 Press the arrowhead beneath the word More Pressing the arrowhead beneath Pswd would lead you into a sub menu that allows you to password restrict access to the keypad functions Pressing the arrowhead beneath the word Reset would allow you to reset all parameters to their factory default values STEP 15 Press the arrowhead beneath the word Exit This will exit you out of the configuration menus and return you to the normal display window Note that communication via the USB or RS 232 ports is disabled while the configuration menu is active so make sure to exit the configuration windows before trying to run the BE2x00 Virtual Instrument so
72. exponential growth rate contant The data field is comprised of an unsigned integer16 Any value that is below the Averaging Time Constant see the A command is set to the Averaging Time Constant for windowing Allowed values are 60 120 240 480 960 and 1920 The default value is 480 Key Slope Window 1 Byte 2 Bytes W 0x57 MSB LSB Sending an empty packet W command will return the w message displaying the current setting 88 Key Slope Window 1 Byte 2 Bytes w 0x77 MSB LSB 15 0x40 Base Unit Embedded Software Version This command reads the version of the embedded software that exists in the Base Unit This data is sent automatically when the device is first powered up The data field is comprised of a 32 bit floating point value The only format for the command is an empty command Key 1 Byte 0x40 Sending an empty packet command will return the 2 message displaying the current setting Key Base Unit S W version 1 Byte 32 bits 2 0x32 LSB MSB 16 0x23 Base Unit Serial Number This command reads the Base Unit Serial Number The field data is represented as a 32 bit unsigned integer Sending an empty packet command will return the 3 message displaying the currently set values Key
73. ff password protection by sending a l byte value for a Q command Sending a 0x00 turns password protection Off Sending a 0x01 turns password protection On Sending a 0x02 resets the password to 1 1 1 1 1 1 and turns Off password protection Sending a 0x03 resets the password to 1 1 1 1 1 1 and turns On password protection Note that the values for 1 1 1 1 1 1 are represented in hex as a six element array of 0x31 Key Q Command 1 Byte 1 Byte Q 0x51 66 99 Sending an empty packet Q command will return the q message displaying the currently set status Key q Value 1 Byte 1 Byte q 0x71 Valid return values are locked 0x00 or unlocked 0x01 This command is only useful for BE2100 and BE2400 base units in the case of the BEIUSB adapter no keypad access is provided so this command serves no purpose 9 R 0x52 Range for Analog Output Sets the range for the analog output terminals of BE2100 and BE2400 Base Units Values are represented as 8 bit unsigned integers Allowed values are 0 5 with a default value of 4 The table below shows the correspondence between the integer representation and the range value Integer Representation Range Value 85 0 0 01 1 0 1 2 1 0 3 10 0 4 100 0 default 5 1000 0 The first byte is the range for Analog Out 1 The second byte is the range for Analog Out 2 BE2100 Base Units only
74. ftware Now that you have a basic familiarity with the operation of the keypad interface you may want to experiment with other features Alternatively a detailed step by step description of the keypad features is available in Appendix II Once you have selected settings that are best for your application you may wish to prevent accidental changes to these settings by turning on the password protection feature Introduction to the BE2400 Base Unit Functions If you are using a BE2100 base unit or a BEIUSB adapter or plan to operate the instrument solely through the Virtual Instrument software you can skip this section However if you wish to operate a BE2400 base unit in stand alone mode without a computer the following tutorial may be helpful This tutorial will give you familiarity with the basic functioning of the keypad interface by leading you through the steps required to modify one parameter the sensor response time STEP 1 Press any of the 4 arrowhead keys on the front of the Base Unit The display should now look like Conf i gurati on Menu 1 cr jo Loc Mor E xk ii A The Scrol 1 and Lock options on the BE2400 base unit provide the ability to scroll the display through all available sensors at 5 second intervals or to lock the display to one particular sensor The factory default setting is scrolling mode Press the More butto
75. g the user calibration units e g mg L e coli Default setting is Cal Units Non printable ASCII characters are ignored Key User Cal Units 86 1 Byte up to 11 ASCII Bytes T 0x54 yas Sending an empty packet T command will return the t message displaying the current string Key User Cal Units 1 Byte up to 11 ASCII Bytes t 0x74 D The last byte is to be used for the null termination character so the maximum useful string length is 10 characters Note The response string contains 1 more character than the command string this last response character should be ignored 12 U 0x55 User Calibration Coefficients Six 32 bit floating point values and one unsigned integer offset linear coeff quadratic coeff cubic coeff MinX and MaxX and Transform Method Min and Max X are the minimum and maximum values of the Sensor Measurement Results used in generating the calibration coefficient These values are useful for determining whether the calibration is being extrapolated beyond the range of the calibration data Transform Method is either linear value 0 or log value 1 Defaults are defined as follows 0 00 1 00 0 00 0 00 0 00 0 00 Key Offset Linear Coeff 1 Byte 4 Bytes 4 Bytes U LSB MSB
76. hat is empty or that contains water or transparent media a sensor reading of less than 0 1 Bug Units is desired but this is not always possible in small e g 1L and less and crowded bioreactors Once the best sensor location has been located and the sensor is tightly secured return the sensor averaging time constant to its normal setting the recommended setting for most applications is 2 minutes Important Reminders A Do not expose the sensor to high temperatures A Remove the sensor during any type of bioreactor autoclave cycle 25 Operation Your BE2100 sensor operates in the near infrared portion of the optical spectrum at 850 nm and is therefore invisible or only slightly visible to the human eye You will not readily observe light emanating from the sensor Normal operation is determined from the display on the Base Unit or the Virtual Instrument software When operating a BE2100 sensor connected through a BE2100 or BE2400 base unit changing the configuration settings and observation of the biomass readings can be performed either in stand alone mode with the keypad and display provided on the base unit or through the Virtual Instrument software when the base unit is connected to a personal computer Sensors connected through a BEIUSB adapter are operated solely via the Virtual Instrument software The BE2100 sensor results are reported in raw Bug Units baseline corrected Bug Units or User Calibrated units
77. he back of the sensor you can bring it closer to the bioreactor the strap tension is insufficient In this case open the latch on the back of the sensor cinch up the free strap end and close the latch again We strongly discourage the use of tapes or adhesives of any kind as they tend to foul the optical surfaces Please contact BugLab at 925 208 1952 or info buglab com for further advice if you are having trouble 24 STEP 3 Inspect the attached sensor to be sure that it is firmly and securely attached to the bioreactor Readjust the mounting strap as required to achieve a firm fit Note a few extra minutes adjusting the sensor for excellent fit will ensure especially reproducible results STEP 4 OPTIONAL STEP RECOMMENDED FOR CROWDED BIOREACTORS Temporarily set the sensor averaging time constant to 0 and observe the reported sensor reading For sensors connected via a BE2100 or BE2400 base unit this can be performed in stand alone mode as already described For sensors connected via a USB adapter the BE2x00 Virtual Instrument Software is required for configuration of and data acquisition from the sensor In this case install the software set the time constant and begin data acquisition following the instructions for operating the Virtual Instrument Software provided in Section II of this manual Re adjust the position of the sensor on the bioreactor to minimize the reported sensor reading Ideally on a bioreactor t
78. in Local Mode when powered on While the keypad configuration menu is active the keypad access will remain in Local Mode until the configuration menu has been exited 30 by the user For this reason you should make sure to exit out of the configuration menus before attempting to start the Virtual Instrument software Display Scrolling BE2400 Base Units only When operated in local mode without a computer the BE2400 display can be set to scroll through all attached sensors or to lock onto 1 particular sensor see Introduction to the BE2400 Base Unit Functions When operated in remote mode the scrolling mode is controlled through the Device Configuration table The last column in the Device Configuration table controls which sensor are included On or excluded Off from display scrolling Password Locking and Unlocking When operated in local mode the BE2100 and BE2400 base units have a password protection feature that can be turned on to prevent inadvertent changing of configuration settings Password protection can be turned On via the keypad by selecting the Pswd option Setting a 6 digit password Set and then turning password protection On Once the configuration menu has been exited subsequent entry into the configuration menus will first require password entry The BE2x00 Virtual Instrument software provides a means of Unlocking the password restriction if the password has be lost or forgotten Thi
79. isplay brightness screen Current bri ght nes s X def aul t EIB P a s s lw r id Prot ec tt o On S e it Baek On Pressing the On button toggles between the On and Off states of password protection The default setting is Off When password protection is On keypad entry into the configuration menus is password protected as shown below Ent er password 1 2 BY Al AJ Al A 74 If the password is not successfully entered within 10 seconds the display reverts to Normal operation If you attempt to turn on password protection without first setting a password the following screen is displayed for 3 seconds followed by a return to the Password Protect menu Pas s wor d mu js t be S e it bef of e selecti ne On If you are locked out of keypad access the BE2x00 Virtual Instrument software provides a command for turning Off the password protection see Working with the BE2100 and BE2400 Base Units Password Locking and Unlocking ii Set Pressing the Set button brings up the following screen Ent er mn e w 6 di i tt IP W 1 2 3 4 If 6 digits are not enter
80. ked due to remote operation by a PC line 1 will display Remote Operation If errors or warnings are detected then the error warning message will be displayed 2 seconds duration alternating with the Remote Operation message 3 seconds duration 2 Line 2 Sensor readings a Ifthe Cal setting is off K IX kK x Bu g Uni t s b Ifthe Cal setting is on The display format and units will be determined by the user e g g L E coli The monitor will poll the sensor approximately once per second and update the screen with the latest information received The letter C will appear at the end of the second line of the display when baseline correction has been turned On On BE2400 base units there is the option of scrolling the display between all attached sensors or locking to the display to one particular sensor When in scrolling mode the results for each sensor are sequentially displayed for approximately 5 seconds each 66 iv Error and warning messages 1 Warnings a If ambient light is higher than desirable but is not preventing a valid measurement then line 1 will read High Ambient Light and line 2 will continue to report the quantitative result If the calibration switch is set to the On position and a user calibration has been stored but extrapolation is required in order to report the result line
81. l as being displayed in a text box to the left of the graph for future reference Note Like the sensor data the event data is saved to file as soon as it is generated The filename into which the event data is saved is the same as the sensor data file except the extension is evt instead of bug Editing Annotations Annotations can be edited by double clicking on cells within the event table and typing in a new value or text string Modifications made to the event table are saved to file as soon as you finish typing them in and hit the enter key or exit the table cell you were editing Events may also be deleted by right clicking the mouse over an annotation and selecting Delete Annotation This action will result in removal of the annotation marker from the graph deletion of the event from the event table and deletion of the event from the saved event file 48 Simultaneous Data Collection from Multiple Sensors Following the same procedures described above data acquisition can be initiated on up to 6 BE2100 sensors simultaneously If a new BE2x00 device has been connected since the last search was performed select the Search Again button within the BE2x00 Device window top portion of the screen Performing a new search will not interrupt data acquisition on devices that are already active Data from multiple sensors can be overlaid and viewed by selecting the All Sensors tab The data type that
82. llowing screen for 3 sec after which the screen returns to the main Communication select screen Curr ent acti Wwe c o m m XXXIX X def aul t USB where XXXXX is either USB or RS232 Disp If the button below Disp is pressed the Display Brightness menu is shown Pswd If the button below Pswd is pressed the Password Protection menu is displayed Display bri ght mnes s Set Hel Show Bac k Set Pressing the Set button allows you to toggle between the allowable selections for the display brightnes 1 2 3 and 4 3 is the default setting as follows Bri ght mess Il ewe ll X Baek 3 Selecting the or keys respectively raises or lowers the brightness level and changes the displayed value of brightness X 1 4 If the display is already at its maximum setting pressing the key has no effect likewise for the key when the display is already at the minimum brightness setting Selecting the Back button brings up the main Display Brightness screen Show Selecting the Show key brings up the following screen for 3 sec after which the screen returns to the main D
83. ly active 39 T 8E2x00 Virtual Instrument w whenever USB power is sensed When communicating via RS 232 make sure that the USB cable is unplugged STEP 2 Connect the RS 232 or USB cable between your Base Unit or BEIUSB Adapter and your computer If this is the first time you are connecting the instrument to the computer via USB you should see a Windows Installing Device Driver Software message pop up If so wait until a second message pops up saying Your device is ready to use before proceeding to the next step Time hrs Value _ Units _ Event Description BE2x00 Devices Biomass Readings Software Settings Device Configuration Device Calibration Calibrated f Pito AB Time Hrs ajasa OD 600nm HY nn 0 01 0 02 0 03 0 04 0 05 0 06 0 07 0 08 0 09 0l Time Hrs BE2x00 Virtual Instrument software main window STEP 3 Launch the BE2x00 software In Windows 7 and earlier select Start All Programs BugLab BE2x00 BE2x00 Virtual Instrument exe In Windows 8 Go to the start screen right click over an empty space and then select All apps lower right hand corner of the screen Scroll through the applications until you find BE2x00 Virtual Instrument Click once on the application STEP 4 When the program is started up a search is automatically initiated f
84. ly attached to the sensor see step 2 above press the button labeled Start Sensor Check and then press Start Low Cal Cup Meas Step iii The instrument will make 25 measurements lasting about 1 second each on the low calibration cup The standard deviation of the 25 measurements is used to assess stability If the measurements were unstable the message The Low Cal Cup measurement was unstable will be displayed If this occurs make sure that the front faces of the sensor and calibration cup are clean and dry and that the calibration cup is securely attached to sensor see step 2 above You will not be allowed to proceed to measurements with the high calibration cup measurements until stable measurements on the low calibration cup have been collected Step iv Once stable measurements on the low calibration cup have been collected the control button will read Start High Cal Cup Meas Replace the low calibration cup with the high calibration cup Make sure the cup is clean and securely attached to the sensor in the same manner as for the low calibration cup see step 2 above Press the button labeled Start High Cal Cup Measurement then the OK button in the pop up screen that appears Step v 25 readings will now be collected on the high calibration cup If the readings were unstable the message The High Cal Cup measurement was unstable will be displayed If this occurs make 22 sure th
85. ly suggest more It is recommended that calibration points spanning the lowest and highest biomass are recorded Collection of calibration points during the exponential growth phase prior to stationary phase usually results in the most successful calibration STEP 4 Press STOP when a full calibration set has been collected Editing Generating and Saving a Calibration STEP 1 Once you have completed a fermentation run during which you collected calibration samples select Cal Window within the sensor graph window for which you want to generate a calibration The User Calibration Window appears The data table on the left side of the screen is automatically populated with the calibration samples from the Event List currently loaded If you wish to read in the calibration samples from a different file select Read from Event File Alternatively if you wish to bring up a previously saved calibration file select Read from Cal File 50 STEP 2 The data table contains four columns Event Raw Sensor Baseline and Calibration The Sensor column contains the data reported by the BE2100 sensor at the time at which the calibration sample was extracted The Baseline column contains the baseline value that was used to collect the BE2100 sensor data The Calibration column contains the matching off line reference data that will be used to generate the calibration If you have not entered any
86. n to access all other configuration options The BE2400 base unit display should now look like Sel ect sensor number 1 2 3 4 A A Note if less than 4 sensors are plugged into the BE2400 base unit only those sensor numbers that are plugged will be shown Note If you are not the 1 user of the instrument and a prior user has turned on password protection before you can reach the above configuration menu you will be prompted to enter a password You may wish to consult the prior user to obtain the password Alternatively if the password has been lost or forgotten the password may be reset via the Virtual Instrument software If this is the case skip this section for now and come back to it later after you have installed the user interface software and reset the keypad password Select the arrowhead button below one of the sensor numbers Note In this exercise you will only be changing the time response for the sensor number that you select If you wish to change settings for more than one sensor each sensor attached to the BE2400 base unit must be individually configured The BE2400 base unit display should now look like Conf i gurati on Menu 2 a s e Ca l Mor E x ji A Pressing the arrows below the words Base or Cal would lead you respectively into sub menus controlling the Baseline and User Calibration features Pressing the
87. ng data acquisition using the Virtual Instrument program such changes are applied prospectively The Data Viewer program allows you to apply the calibration uniformly across the entire data file In other situations you may wish to see the effect of applying different calibrations to the same data set STEP 1 From within one of the File tabs press the Open button select the file you want to work on and then press OK STEP 2 From within the same File tab press the Adjust Cal button The Set Calibration Retrospectively vi window will open STEP 3 Set the Calibration Update Method control to one of the four settings Set Manually If you select this option you should type the new calibration settings directly into the Calibration Settings control This option is useful in circumstances in which you have already performed a calibration fit in an external program e g Microsoft Excel Retrieve from Cal Table If you select this option the settings that are currently in the User Calibration table in the top half of the program window are written into the Calibration Settings control Retrieve from Cal File This option is useful if you have previously saved a calibration file and want to apply the same calibration settings to the currently open data file Open Cal Window Selecting this option will open the User Calibration window This is the same
88. ng outside of its normal range warning or error messages will be displayed See Part I Working with the BE2100 and BE2400 Base Units in this manual for more details on the types of warning or error messages that may be displayed The Display Mode selector to the upper left of the graph allows you to select the data that is displayed on 43 the graph either Raw Baseline Corr Calibrated or Growth Rate Setting the Baseline The Baseline setting is used to subtract a constant offset from the raw and or calibrated sensor data For example when running in Calibrated mode using a calibration file that converts the raw Bug Units into OD units setting the baseline is just like zeroing your spectrophotometer with a reference measurement In a typical experiment the baseline is set near the beginning of a fermentation run just prior to inoculation as described below To set the baseline the user normally specifies a range start and end point as selected by right mouse button and the baseline is determined by averaging over the specified range Alternatively the baseline can also be set manually The following step by step description shows how to use either method STEP 1 While running an experiment the Baseline can be set by positioning the mouse arrow over the graph window at the time position at which you would like to start baseline averaging and clicking on the right mouse button From the drop down lists that appear ch
89. nit The BE2100 base unit display should now look like Conf i gurati on Mendu 1 ass e Cal Mo r E x i Pressing the arrowhead below the words Base or Cal would lead you respectively into sub menus controlling the Baseline and User Calibration features Pressing the arrowhead beneath the word Exit would return you to the normal display screen Note If you are not the 1 user of the instrument and a prior user has turned on password protection before you can reach the above configuration menu you will be prompted to enter a password You may wish to consult the prior user to obtain the password Alternatively if the password has been lost or forgotten the password may be reset via the Virtual Instrument software If this is the case skip this section for now and come back to it later after you have installed the user interface software and reset the keypad password STEP 2 Press the arrowhead beneath the word More The display should now look like Conf i gurati on Menlu 2 Chk AW e Mor Exi Pressing the arrowhead below the word Chk would lead you into the Sensor Check function This function allows you to verify sensor performance and if necessary update calibration coefficients using measurements made on the Low and High calibration cups
90. nnect the USB cable from the BE2x00 instrument and the computer b Under the Driver tab select Uninstall c Inthe Uninstall window that pops up click on Delete the driver software for this device and hit OK Install the driver a If you haven t done so already disconnect the USB cable from the BE2x00 instrument and the computer b Go to the root directory where the BE2x00 software was installed By default this is C Program Files BugLab BE2x00 c Within this folder double click on the file named CDM v2 12 00 WHQL Certified exe d In the pop up window answer Yes to allow changes to your computer A window entitled FTDI CDM drivers should open Select Extract The Device Driver Installation Wizard should automatically start Select Next and wait for a confirmation that the driver was successfully installed Select Finish to exit out of the Installation Wizard Plug in the BE2x00 instrument a Make sure a BE2100 sensor is connected to your base unit or BEIUSB adapter b If you are working with a BE2100 or BE2400 base unit make sure it is plugged in turn on the base unit and wait for the sensor to be detected c Connect a USB cable to the BE2x00 instrument and to the computer b You should see a windows message Installing device driver c Wait until a new message appears Device driver software installed successfully Start up
91. nored Key Baseline Correction User Calibration On Off On Off 1 Byte 1 Byte 1 Byte O 0x4F Sending an empty packet O command will return the o message displaying the currently set values Key Baseline Correction On Off User Calibration On Off 1 Byte 1 Byte 1 Byte o Ox6F 84 8 Q 0x51 Base Unit Password Settings You may wish to set up password protected access to the keypad functions on the Base Unit When in the locked state entry to the Base Unit functions is enabled by entering a six number value via the keypad Only values from 1 to 4 are valid for each number If the six numbers ASCII entered on the keypad match the currently stored password the system enters an unlocked state until the user exits the configuration menu If the numbers entered do not match the currently stored password access to the configuration menus is denied and returns to the normal display The value of the Base Unit password is stored in the Base Unit The default password is 111111 In addition to changing the password via the keypad on the Base Unit the password may be set through the Q command Key P Valuel P Value2 P Value3 P Value4 P ValueS P Value6 1 Byte 1 Byte 1 Byte 1 Byte 1 Byte 1 Byte 1 Byte Q 0x51 The Q command may also be used to reset and turn On or O
92. nsor female 4 wire threaded Analog Output 4 20mA 5000 max or 2 10V using provided 500 Q resistors Resolution 12 bits 0 004 mA or 2 mV Range Settings 6 settings logarithmically spaced 0 01 1000 Number of Outputs 4 and screw terminal positions for sensor signal and exponential growth rate Digital Output RS 232 DB9 and USB Communications Cable optional Connectors DB9 M F or USB A B Length Standard 2m 6 Custom Base Unit Physical Overall Width up to 15m 50 15 9cm 6 2 Overall Length without connectors 13 2cm 5 2 Overall Height without feet Base Unit Environmental Operating Temperature 8 2m 3 2 0 to 40 C 30 to 100F Operating Environment BE 2100 62 dry location use only BE2400 Base Unit Features Base Unit Features Access to individual sensor and base unit settings through interactive Keypad Password control optional of keypad access Lighted display with variable brightness control Digital USB and RS 232 and four analog 4 20mA or 2 10V outputs User Interface Software Features Real time graphical and numerical display for each sensor Event marking on graph both pre defined and user defined Baseline setting and subtraction User calibration of sensor output to any reference units Acces
93. of cell growth Cell lysis results in a dramatic change in the average particle size Once significant cell lysis has begun such as typically occurs during the stationary phase of cultures there will no longer be a linear relationship between biomass and optical measurements of scattering by either conventional spectrophotometry or by the BE2100 sensor Attempts to apply a non linear fit to accommodate more than one particle size at a time such as happens due to cell lysis is not likely to be reliable because there is insufficient information to distinguish between changes in particle size vs changes in number of particles For this reason when generating new calibrations we recommend only using data collected prior to the transition between logarithmic and stationary phases of cell growth What about the effects on measurement accuracy of vessel size shape wall thickness and material The BE2100 sensor instrument is designed to provide accurate results over a wide range of vessel types This is accomplished by limiting the optical measurement volume and restricting the allowed geometry between the vessel and instrument Through a combination of these methods the BE2100 sensor has been demonstrated to provide high accuracy lt 15 over a wide range of vessel types and wall thicknesses up to 13 mm References 1 U S Patent 8 603 772 Particle Sensor with Wide Linear Range 2 US Patent 7 100 462 Self Adjusting Sensor
94. omputer goes into sleep mode This will cause the BE2x00 Virtual Instrument software to lose it s connection with the sensor STEP 2 If you are working with a BE2100 Base Unit select the communication port type on the base unit as follows Step i Press any of the 4 arrowhead keys on the front of the Base Unit Step ii Press the arrowhead beneath the word More three times The top line of the display should now read Configuration Menu 4 Step iii Press the arrowhead below Comm The top line of the display should now read Communication Select Step iv Press the arrowhead below USB or RS232 depending on the type of communication port you are using Your new selection along with the default value will be displayed briefly on the screen Step v Press the arrowhead beneath the word Exit Note 1 Virtual instrument communication with the base unit is locked out while the configuration menus are active Make sure you have returned to the normal screen before proceeding to the next step Note 2 When USB is selected at the communication type RS232 communication is still functional as long as a USB cable is not also connected However when RS232 is selected as the communication type USB communication is disabled Note 3 If you working with a BE2400 Base Unit you will notice that there is no Comm setting On BE2400 Base Units USB communication is automatical
95. on Example Appendix V Trouble Shooting 57 59 66 76 90 92 INTRODUCTION This User Manual describes the operation of the BE2100 non invasive biomass sensor The sensor measures biomass in liquid cultures using laser optical reflectance at 850 nm The optical reflectance is measured through the vessel wall of glass fermenters or through transparent port windows provided in stainless steel bioreactors The sensor must be placed below the liquid air interface in order to provide accurate measurements Calibration tools are provided so that results can be reported in whatever biomass units are desired such as dry cell weight g L optical density OD or cell density cells mL Specific calibration to the particular type of organism being measured is recommended Tools are also provided for baseline correction of media reflectance as measured in the absence of biomass There are currently three options for connecting to the BE2100 sensor as depicted in the Figure on the next page these different options are collectively referred to in this manual as the BE2x00 instrument In the standard configuration the BE2100 sensor is connected to a BE2100 base unit and operated either in conjunction with a personal computer PC or in stand alone mode The B2100 base unit provides a connection for one BE2100 sensor The sensor configuration may be set up either through a keypad interface or via the provided Virtual Instrument software Bioma
96. oose Create Annotation gt gt Baseline Start STEP 2 The Add Annotation Window now becomes active Under the box labeled Positioning Method notice that the selected method is Cursor This means that the Baseline Start will be marked at the position at which you right clicked the mouse Alternatively if you wish to start the baseline averaging at the most recently collected data point select Add to End Click on the Add Annotation button at the bottom left of the pop up window STEP 3 Notice that a red marker and number have been added onto the graph indicating the start of baseline averaging An event has also been added to the Event List located to the left of the graph Continue the data acquisition process until you would like to define the end point for baseline determination Position the mouse arrow over the graph window at the time position at which you would like to end baseline averaging and clicking on the right mouse button From the drop down list that appears select Create Annotation gt gt Baseline End STEP 4 The Add Annotation Window again becomes active Leave the Positioning Method set to Cursor Position if wish 44 to mark the Baseline End at the point where the mouse was right clicked Alternatively if you wish to start the baseline averaging at the most recently collected data point select Add to End Click on the Add Annot
97. or all connected BE2x00 devices For those BE2x00 devices that are identified all configuration settings are read from instrument memory and put into program memory This process may take several minutes but once complete a list of all available devices will be shown in the BE2x00 Devices page at the top of the screen All configuration settings as read from instrument memory can also be viewed in the Device Configuration and Device Calibration pages 40 The bar separating the top and bottom portions of the window can be adjusted over a limited range by clicking and holding the mouse on the bar and moving the bar to the new desired position before releasing the mouse again Notice that Sensor Tabs at the bottom of the page are enabled according to the number of BE2100 sensors that were identified The sensor number on the sensor tabs corresponds to the sensor columns in the configuration tables at the top of the page If you wish to change the order in which the sensors are numbered you can do so by entering the desired order into the New Order column of the BE2x00 Devices page and then selecting the Rearrange button The sensor numbering will be automatically updated in all configuration tables and in the Sensor Tabs at the bottom of the page STEP 5 Select the Software Settings tab at the top of the page The Sampling Interval column in the table determines the frequency with which
98. orbance and scattering As a result a highly linear relationship is maintained between biomass concentration and the measured optical reflectance However it is also important to realize that for biomass containing strongly visible light absorbing chromophores e g photosynthetic algae the chromophore absorbance may affect the agreement between a conventional OD measurement and the result reported by the BE2100 sensor The OD measured in the visible range by a spectrophotometer will be influenced by both chromophore absorbance and cell scattering whereas the OD reported by the BE2100 sensor will be based only on cell scattering In such situations if the relationship between chromophore concentration and biomass is relatively fixed it still may be possible to 34 generate a strong correlation between OD measured by conventional methods and that reported by the BE2100 sensor However it is important to be aware that changes in chromophore concentration that are not accompanied by biomass change would not affect the result reported by the BE2100 sensor but would skew the results determined by conventional visible spectrophotometry Another important consideration when comparing BE2100 sensor results with conventional spectrophotometry is the optical design of the spectrophotometer Most spectrophotometers are designed to make accurate measurements of absorbance but not scattering A determination of chromophore absorbance requires only a
99. orrection Baseline correction provides a means of subtracting off signals emanating from reflectance sources in the bioreactor that are not of interest For example in a typical application the baseline will be measured just prior to inoculation thereby subtracting off the contribution of media constituents from the reported Bug Units This function is similar to zeroing of a spectrophotometer using only medium prior to performing an OD measurement Before collecting a new baseline it is important to establish that the signal is stable Viewing the signal in graphical format such as provided in the Virtual Instrument software can be helpful for this purpose If the baseline appears to be wandering excessively it can sometimes be helpful to temporarily reduce the sensor averaging time constant to 0 while adjusting the sensor position and or the bioreactor conditions see Step 4 of Setting Up for a Bioreactor Run The baseline correction function can be accessed both via the keypad interface on BE2100 or BE2400 Base Units or via the Virtual Instrument Software Use of the keypad interface is described here For the Virtual Instrument Software refer to section II E 2 of this manual In the following it is assumed that the sensor has already been attached to the bioreactor and that a stable reading has been achieved STEP 0 Press any of the 4 arrowhead keys on the front of the Base Unit If you are using a BE2400 base
100. ow the graph The coefficients are listed from lowest to highest polynomial order offset linear quadratic and cubic STEP 6 If you are satisfied with the fit select the Accept Fit button You will be prompted to choose a path and name for the calibration file to be saved Following file saving the new calibration is automatically written into sensor memory The calibration coefficients will be remembered across power cycles The instrument can now be run in calibrated mode without the need to be hooked up to a computer Running in Calibrated Mode Now that you have saved a calibration into sensor memory if you wish to collect calibrated data you simply need to turn On calibration Calibration can be turned On within the Virtual Instrument software from the User Cal column of the Device Configuration table tab at the top of the screen Alternatively the keypad interface on the Base Unit can be used to turn User Calibration Cal On or Off Note When running in calibrated mode the Error Code Interpretation column of the Biomass Readings table will display a warning message if the raw sensor data falls outside the calibrated range In the event of this condition the message Warning Extrapolating Beyond Calibration will be displayed 52 DATA VIEWER Opening Viewing and Resaving Data Files A separate BE2x00 Data Viewer program is provided for displaying previously acquired data Th
101. pectively data points that were acquired prior to setting the baseline are not affected by the new baseline When a new baseline is set via this create annotations method the baseline correction is automatically turned On Baseline correction can also be manually turned On or Off within the Device Configuration table If you are using a BE2100 or BE2400 Base Unit the display will include a C at the end of the second line when baseline correction is On Modifying the Data Acquisition Window Several options are available to allow you to customize the manner in which the data is displayed in the Data Acquisition Window 1 You can change the scaling of the graph axes 45 By right clicking over the graph the autoscaling of both the X and Y axes can be turned on or off When auto scaling is turned on the BE2x00 software will select axis limits that best display all of the data collected since the data acquisition was begun When autoscaling is turned off you can change the values on the axes by clicking one or more of the extreme values of the grid labels and changing their values The pan depicted as a hand and zoom depicted as a magnifying glass features at the left bottom of the graph can also be used to change the range of the graph that is displayed If you pan or Zoom while autoscaling is on the graph will rescale whenever new data is added to the graph that falls outside of the current window Regardless of the mode
102. ry On Off Baseline This is a command from the Base Unit BEIUSB adapter or PC to the sensor to start restart end or query the baseline status Key K Value 1 Byte 1 Byte K 0x4B 80 K Value Meaning 0x01 start baseline gathering 0x00 stop baseline gathering OxFF cancel baseline gathering If the sensor is not gathering baseline data setting the K value to 0x01 will command the sensor to start gathering a baseline reading Setting the sensor to baseline gathering mode will put the sensor into a mode where the averaging time constant is approximately 1 sec If the sensor is currently gathering baseline data the sensor is put in a sensor busy mode This prevents the sensor from sending messages or receiving commands for the duration of the progress with the exception of an interrupt baseline command An error code is sent back with the value of system busy If the baseline is gathering data setting the K value to 0x00 will command the sensor to stop gathering a baseline reading Upon completion of gathering a baseline the baseline B b value is updated and the Averaging Time Constant is restored to the original value Note that the B message is not automatically sent upon completion of calculation of the baseline If the baseline is not gathering data setting the K value to 0x00 will have no effect If the baseline is gathering data setting the K value to OxFF will command the sensor
103. s 2 Calibration to external reference standards via user interface software Sensor Electrical DC Power In 5V 90 mA Certifications CE marked Tested for compliance to EMC standards EN55011 and EN61000 and safety standard EN 61010 Sensor communication with base unit or Serial digital protocol BE USB available on request Sensor Cable Connector BE2100 sensor male 4 wire threaded Length Standard Extension cable avail as accessory Sensor Environmental and Safety Operating Temperature 0 to 50 C 30 to 120F Environmental Seals Yes Splash Proof Laser Product Classification 1M 59 Sensor Physical Front Face Width 15mm 0 59 Front Face Height 41mm 1 61 Overall Length excluding latch 100mm 3 9 Active Optical Window length x width 25mm 1 0 x 6mm 0 23 Min Vessel Diameter 5 3cm 2 1 Max Vessel Diameter std Strap 34cm 13 5 Max Vessel Diameter extended Strap Unlimited Max Window Depth 31mm 1 2 Functions with Flat Surfaces Yes External Materials Body Aluminum Gasket PVC Filter Acrylic Strap Nylon Latch Stainless Steel Fasteners Stainless Steel Cable Strain Relief Polyurethane BE2100 Sensor Dimensions 30 mm C 15mm 94mm 44 4mm 41 mm ADJUSTABLE OTO 31 mm 60 BE2100 Base Unit Features Base Unit F
104. s accomplished within the Device Configuration table by selecting the table cells in the column labeled Base Unit Password Lock Status Four options are available Unlock ed Turns Off password protection Lock ed Turns On password protection Reset and Unlock Turns Off password protection and resets the password to 111111 Reset and Lock Turns On password protection and resets the password to 111111 Warning Messages It is possible to overwhelm the BE2100 sensor if ambient light conditions are extremely high You may see the warning shown below occasionally If it is infrequent it may be safely ignored The BE2100 always becomes vastly more resistant to high ambient light as your organism reaches higher optical density If the sensor is exposed to direct sunlight halogen 31 lamps at close range or similar conditions the following warning condition may be posted High Ambient Light 248 1 Bug Units The signal displayed is accurate but the user is encouraged to lower ambient lighting conditions If User Calibration is turned on but the present measurement is outside of the calibration range the following warning is displayed Extrapolating Cal 248 1 Bug Units Error Messages If the ambient light conditions are so high as to prevent accurate measurements the BE2100 will post the following error message Signal Saturated You must lower ambient lighting conditions in order to measure
105. s to all sensor and base unit settings User Interface Software Requirements Windows XP Vista 7 8 82 or 64 bit Operating System Minimum of 256 MB of RAM Minimum 200 MB free hard disk space 1024 by 768 resolution or higher video adapter Available USB or 9 pin RS 232 COM port Microsoft compatible mouse CD reader required only at time of software installation F i mad a m 147 mm 132 mm Jt A aor I 1 8mm Fa x r 63 BE2400 Base Unit Specifications Base Unit Electrical DC Power In Both US and International plug adapters available 6V 1A Certifications CE marked Tested for compliance to EMC standards EN55011 and EN61000 and safety standard EN 61010 Sensor Input Up to 4 BE2100 sensors Connector BE2100 sensor female 4 wire threaded Analog Output 4 20mA 5000 max or 2 10V using provided 500 Q resistors Resolution 12 bits 0 004 mA or 2 mV Range Settings 6 settings logarithmically spaced 0 01 1000 Number of Outputs 8 and screw terminal positions for each sensor Digital Output RS 232 DB9 and USB Communications Cable optional Connectors DB9 M F or USB A B Length Standard 2m 6 Custom Base Unit Physical Overall Width 15 9cm 6 2
106. sensor this step should provide the final pressure needed to firmly seat the sensor against the calibration cup STEP 3 Start the Sensor Check function A VIA THE KEYPAD INTERFACE ON BE2100 OR BE2400 BASE UNITS Step i Press any of the 4 arrowhead keys on the front of the Base Unit Note 1 If the Virtual Instrument software is currently communicating with the base unit as indicated by Remote Operation on the top line of the display keypad access is locked out You will either need to suspend communication or perform the sensor check from within the Virtual Instrument software described below under B Note 2 If you are not the 1 user of the instrument and a prior user has turned on password protection before you can reach the above configuration menu you will be prompted to enter a password You may wish to consult the prior user to obtain the password Alternatively if the password has been lost or forgotten the password may be reset via the user interface software If this is the case skip this section for now and come back to it later after you have installed the user interface software and reset the keypad password Step iia BE2400 base units only Press the arrowhead beneath the word More The top line of the display should now read Select sensor number Step iib BE2400 base units only Select the sensor number for which you want to run the Sensor Check function
107. single message of sensor disconnected to the serial or USB port 5 Data Protocol Message Data 1 Command keys recognized by all BE2x00 devices are shown on the left side of Table 1 The response keys to these commands are shown on the right side of Table 1 The commands are all ultimately received and responded to by a BE2100 sensor When sent to a base unit or BEIUSB adapter the commands are automatically relayed to and from the connected BE2100 sensor s Further details of these commands can be found in section 6 below Command keys recognized BE2100 Base Units BE2400 Base Units and and BEIUSB adapters but not by BE2100 Sensors are shown on the left side of Table 2 The response keys are shown on the right side of Table 2 Comands recognized only by BE2100 and BE2400 Base Units are shown in Table 3 Commands recognized only by BE2400 Base Units are shown in Table 4 While the Base Unit menu configuration menu is active activated by pressing any of the 4 keys on the pad any commands sent to the Base Unit will be responded to with the lower case complement of the command plus a 1 byte data packet whose value indicates that the system is busy error code 8 Commands sent to the sensor while it is busy e g in the middle of running the sensor check routine or while it is in the middle of collecting 77 baseline data will likewise be responded to with the lower case complement of the command sent plus a 1 byte data
108. sity The detectors within the BE2100 sensor are sensitive to the intensity of light back scattered from within the glow balls Further the photo sensor is arranged such that there are multiple separation distances between the detectors and light sources This is of critical importance to the BE2100 technique The sensor firmware automatically arbitrates among the source detector pairs selecting that pair with the most linear response to biomass at the particular biomass currently being measured By combining the signals from the multiple detectors due to the multiple light sources the biomass in the liquid culture is determined with a dynamic range that is orders of magnitude larger than immersion transmission techniques Although classical Optical Density OD measurements in a spectrophotometer require dilution in order to accurately determine biomass greater than about 0 5 OD 600 nm the BE2100 sensor is able to determine biomass from 0 1 to 300 OD 600 nm without dilution or any liquid handling The high linearity of the BE2100 sensor allows growth rate to be accurately and rapidly assessed whether or not the sensor has been user calibrated biomass over a very wide range of response One of the advantages of using an infrared laser source for measuring biomass is the avoidance of light absorbance by colored media components and colored vessel materials This allows for the measurement of true scattering rather than a combination of abs
109. sor in the same manner as for the low calibration cup see step 2 above Press the arrowhead beneath the word Ready Step ix 25 readings will now be collected on the high calibration cup If the readings were unstable the message Hi Cup unstable RPT will be displayed If this occurs make sure that the front faces of the sensor and calibration cup are clean and dry and that the calibration cup is securely attached to sensor see step 2 above It will be necessary to repeat the low calibration cup step v above measurement before proceeding again with the high calibration cup measurement If the readings were stable a Pass Fail assessment of sensor performance will be made This Pass Fail assessment is based on a comparison of the present 21 measurements to prior measurements made on the calibration cups during manufacture B VIA THE VIRTUAL INSTRUMENT SOFTWARE Step 0 If you have not done so already follow the steps under Setting Up of the BE2x00 Virtual Instrument Software section II of this manual Step i Start up the software and wait for the initial Device Search to complete In the lower half of the screen select the tab of the sensor for which you wish to run the Sensor Check function Press the Check Sensor button in the lower left of the screen Note The Sensor Check function cannot be run while data acquisition is active Step ii Once the low calibration cup has been secure
110. sor response The set new variable sensor coefficients command should only be sent when stable measurements have been completed on both the Low and High Calibration Cups When the cal cup measurement is interrupted by sending an L value of OxFF the calibration cup measurement data will be cleared from sensor memory with no change to the Variable Sensor Coefficients Sending an empty packet L query to the sensor after sending and L command with an L Value of either the 0x00 clear data or OxXFF interrupt and resume normal mode will result in an l value of 0x00 no data being returned Returned I packet in normal mode Key l Value Measured Combined Stored Result Combined Result 1 Byte 1 Byte 4 Bytes 4 Bytes TP 0x6C MSB LSB MSB LSB When the Sensor Check routine is running in the sensor all commands other than an L command will be responded to with a system busy 1 byte error code 6 M 0x4D Data Request This command is used to request a data packet from either the sensor or the Base Unit Returns the sensor data output The only format for the M command is an empty M command Key 1 Byte M 0x4D Sending an empty packet M command will return the m message along with a data packet The data packet has 6 components 83 1 Error Code 8 bit signed int
111. sor will result in one of 6 I values being returned The returned l value can be used to determine the type of calibration cup that was measured low or high whether the standard deviation of the 10 calibration cup measurements was above unstable or equal to or below stable a threshold value In the case that the 82 measurement was stable the 1 value further indicates whether or not the sensor result when compared to the result computed from the stored Calibration Cup Coefficients had an absolute error that exceeded failed or was equal to or below pass a threshold value If calculation and saving of new Variable Sensor Coefficients is in progress when an empty L command is sent to the sensor a Busy 0x13 L value will be returned to the sender When new Variable Sensor Coefficients have been computed and saved sending an empty L command to the sensor will result in an 1 value of 0x63 being returned When an error is encountered during the calculation or saving of new Variable Sensor Coefficients sending an empty L command to the sensor will result in an 1 value of 0x53 being returned In addition to the 1 Byte 1 value 2 32 bit floats are returned in response to an empty packet L command The first float value is the sensor response in Bug Units measured on the calibration cup the second float value is the stored sen
112. specially when working with complex media that may vary in turbidity from batch to batch baseline correction on the media alone is recommended When a User Calibration is used to transform the Bug Units data into some other reference the transform is applied to the baseline corrected result For this reason it is particularly important to consistently apply the baseline correction when applying User Calibrations 26 The BE2100 can be sensitive to changes in sparge and stir rate particularly at low OD s If possible establish ideal sparge and stir rates and leave them constant throughout the monitored fermentation If changes are required in sparge or stir rate note them The BE2100 baseline may change as sparge or stir rate are changed but it will still precisely track growth The effect of glass wall thickness between different types of bioreactor vessels has been tested and has a nearly negligible effect on BE2100 sensor response in the range of 6 to 13 mm this range encompasses most glass vessels and well as flat glass ports on stainless steel reactors The sensor incorporates a patented automatic mechanism to maintain a constant distance between the optical components and the outside surface of the bioreactor However this mechanism only functions if the sensor is held snugly to the bioreactor For this reason it is critical to ensure that the strap is tightly secured as detailed in Setting Up On a Bioreactor Baseline C
113. ss results are visually displayed on the BE2100 base unit screen and are also available in digital RS 232 or USB and analog 4 20 mA or 2 10 V format The BE2400 base unit offers the same functionality as a BE2100 base unit but with the ability to connect to up to 4 BE2100 sensors A third sensor connectivity option is the BEIUSB adapter This small adapter allows the BE2100 sensor to be plugged directly into a PC With this option all configuration setting and data communications are performed via the PC The BE2x00 Virtual Instrument software provided with the instrument works with all of the above described hardware configurations Configuration of calibration of and data collection from up to six sensors can be simultaneously performed from within the same program window At program start up a search is automatically performed for all connected BE2100 sensors All configuration settings are displayed in tables and the settings can be modified with a simple mouse click Data acquisition is graphically displayed in separate tabbed windows for each sensor Important events that occur during the bioreactor run can be marked directly on the graph and are displayed in a summary table for each sensor A combined graph window also allows the results for all sensors to be overlaid A separate BE2x00 Data Viewing program allows previously acquired data to be viewed manipulated and resaved Detailed instructions for operating both the BE2x00 Virtu
114. standard deviation SD is computed for each of the detector signals 1 If the SD is less than Threshold 1 for all of the signals the base unit displays Cont i n ufe t jo H ii Cal P Y le s IN jo a Ifthe Yes button is selected the routine continues to the next screen b Ifthe No button is selected the Sensor Check routine is terminated and the previous Configuration screen is displayed 2 Else the monitor displays the message Uns tables Repeat Y le s IN jo a If Yes is selected the routine continues back at step c il b If No is selected the routine is terminated and the previous Configuration screen is displayed 71 vi vii viii High Cal Cup Ifthe Low Cal Cup measurement is successfully completed the High Cal Cup screen is next displayed At tach Hi g h Cal Cup Re a d y Heil p B a e k The subsequent High Cal Cup screen sequences will be the same as those shown above for the Low Cal Cup sequence If both the Low Cal and High Cal measurements are successfully completed they will be converted into Bug Units Then the absolute value of the errors for the cal cup measurements are computed relative to the expected val
115. t 1 Serial data is sent in packets 2 The packet starts with a header byte OxSA ASCII character Z 76 ue The second byte in the packet is the length byte Length byte is the total length of the data payload min length of 0 max length of 127 The third byte in the packet is the key byte command and response keys provided in Tables 1 4 The next byte s up to but not including the checksum are the data bytes The data bytes can be zero bytes empty packet or up to 127 bytes of data The length byte described in 4 c above refers to the total length of these data bytes The next to last byte is the checksum The checksum chosen is a mod256 checksum byte calculated using the key byte data bytes and length byte The checksum is formed by adding the hex value of all bytes and applying a modulus 256 to the sum The last byte is the footer byte 0x3C ASCII character lt 4 Packet Send amp Receive Specifications 1 2 3 Each packet has a single message only On Power on of the Base Unit the Base Unit will send a single Base Unit version packet to the serial or USB port If no sensor is connected at Base Unit power on the Base Unit will send a single message of No sensor connected to the serial or USB port On detection of a sensor the Base Unit will send a single sensor version packet to the serial or USB port On detection of a sensor disconnect status the Base Unit will send a
116. t immerse the optical sensor Do not drop or shake the sensor gt B Do not expose the sensor to direct sunlight or high temperatures A Remove the sensor during any type of bioreactor autoclave cycle Q There are no user serviceable parts inside the sensor or the main unit a Do not leave fingerprints or dirt on the optical sensor surface The sensor is a precision optical device It may be cleaned with a light application of commercial window cleaner ethanol or isopropanol and gentle wiping with paper or cloth towels Optical grade lens cleaning tissue is preferred Do not use acetone to clean the sensor Some of the materials used in the sensor will rapidly degrade if exposed to acetone AN Important note to users with more than one BE2100 sensor The calibration cups provided for testing sensor performance are not identical and should be matched with the corresponding sensor The last 2 digits of the serial numbers printed on the sensor and low and high calibration cups should all match each other A Important note to BE2x00 software users Before plugging the BE2100 into your computer make sure you have installed the software first Configure the power settings on your computer to never go into sleep mode Table of Contents Part I BE2x00 Biomass Monitor Part II A Introduction B Getting Started 1 Unpacking the Instrument 2 Configuring the Power Adapter 3 Connecting the Sensor and Base Unit
117. tion the Optical Sensor Head on your bioreactor so that it will get a good view of the liquid culture Most bioreactors are quite crowded with impellers sparge tubes pH probes etc This effect is exacerbated in very small bioreactors Try to find a location relatively free of internal obstructions that would block or reflect emitted sensor light inside of the bioreactor This effect is most important at low biomass levels Your BE2100 sensor will achieve highest immunity to internal obstructions at highest biomass levels near harvest time The BE2100 sensor has maximum sensitivity to objects at a distance of about 2 cm from the front face of the sensor However at very low biomass reflectance from farther objects can sometimes influence the readings 23 STEP 2 Although your BE2100 Optical Sensor Head will adapt to virtually any diameter bioreactor including flat glass viewing ports because of the myriad bioreactor styles that the BE2100 may encounter this step sometimes requires a dry run When mounting to all glass cylindrical vessels select and apply a combination of magnetic sensor adapters so that when the front face of the sensor is depressed the sensor front face can be made flush with the adapters The best combination of adapters for this application is generally the 2 longest adapters 15 and 20 mm stacked on top of each other When mounting to flat recessed ports with a narrow aperture it may necessary to use a
118. to interrupt and cancel the gathering of a new baseline reading Upon canceling the baseline B b value is not updated and the Averaging Time Constant is restored to the original value If the baseline is not gathering data setting the K value to OxFF will have no effect Sending an empty packet K command will return the k message displaying the currently set values Key K Value 1 Byte 1 Byte k Ox6B The returned values are either 0x01 currently collecting baseline data or 0x00 not collecting baseline data 5 L 0Ox4C Sensor Check Function Control This is a command from the Base Unit BEIUSB adapter or PC to the sensor to start end or apply the results of the Sensor Check function The Sensor Check function runs from within the sensor embedded software The purpose of the Sensor Check routine is to determine if the sensor is performing correctly and if necessary reset the internal 81 sensor calibration A one byte value following the L command provides the control for the Sensor Check routine Key L Value 1 Byte 1 Byte L Ox4C L Values Meaning 0x00 Clear data 0x01 Initiate Low cal cup measurement 0x02 Initiate High cal cup measurement 0x03 Use cal cup measurements to set new Variable Sensor Coefficients OxFF Interrupt measurement and resume normal mode Note values not defined above are illegal an
119. ty packet B command will return the b message displaying the current setting Key Current Baseline Value 1 Byte 4 Bytes b 0x62 LSB MSB 3 UJ O0x4A Base Unit Keypad Lockout This command is used to lock J value 0x01 or unlock J value 0x00 access to the Base Unit keypad configuration menus The default is unlocked The state is always reset to unlocked at power up of the Base Unit When a J1 lock base unit command is sent to a BE2400 multiplexed base unit the base unit is automatically taken out of the scrolling state The scrolling state prior to sending the J1 command is saved in memory and automatically restored when the base unit is unlocked Key J Value 1 Byte 1 Byte P 0x4A Sending an empty packet J command will return the j message displaying the currently set value Key j Value 1 Byte 1 Byte J O0x6A When communicating with BE2100 or BE2400 base units it is recommended that the base unit keypad be set to the locked state prior to performing any other communication steps This will prevent conflict between serial communication commands and manually initiated keypad access This command is only useful for BE2100 and BE2400 base units in the case of the BEIUSB adapter no keypad access is provided so this command serves no purpose 4 K 0x4B Start End Que
120. ue If the absolute errors are less than or equal to Thresholds 3 and 4 respectively for the Low Cal and High Cal measurements the monitor displays Slns ot Plats s edh Upda ft e Hel p Bat k 1 Ifthe user selects Update the new offsets and gains are determined based on the low and high cal cup measurements The low cal cup reading on each detector is the offset value for each detector The high cal cup reading on each detector is divided into a reference reading for each detector stored in sensor memory to determine the gain for each detector The new sensor calibration coefficients are then written to memory 2 The Back button allows the user to step back in the menu Otherwise the monitor displays the message Sl ns ot Fai ledh Upda t fe Hel p Bat k The Update and Back functions produce the same result as in the Sensor Passed case section h above d Averaging Time Constant Selection Ave Pressing Ave button displays the Time Constant selection menu li Ti mee Cons t abt Select Set Hel p Show Bac k Set Pressing the Set button brings up the following screen Sel ect Ti me Constant 0 S e c B 0 S
121. ures these cell size differences can be compensated using a single multiplicative factor calibration slope Note that the 10 fold difference in cell diameters of these 2 microorganisms only has about a 2 fold effect on the BE2100 calibration to biomass For this reason minor variations in cell diameter such as are observed in different stages of growth or between different strains of the same organism will have a relatively minor effect on the BE2100 sensor accuracy Note that correlation between biomass and OD measurements performed using conventional spectrophotometry are also cell size dependent and that this cell size dependence is somewhat different than for the BE2100 sensor due to the difference between the optical measurement geometries e g transmission vs reflectance 35 More serious consideration must be given to organisms that do not grow as mono disperse cells e g filamentous growth The relationship between OD whether determined by conventional means or the BE2100 sensor and biomass is non linear for organisms that are not mono disperse in a liquid culture As a result OD can only be expected to provide an accurate measure of biomass for mono disperse liquid cultures For organisms that grow in clusters OD can only be expected to provide a qualitative estimate of biomass Correlation between biomass and BE2100 sensor measurements will generally be highest during the exponential aka logarithmic phase
122. w baseline is briefly displayed on the screen The new value can be viewed again by pressing the Show button STEP 3 If the lower left of the screen shows Off press the first arrowhead key so that the display changes to On Even though the new baseline has been set it will not be applied to the displayed data unless the correction is On STEP 4 Press the last arrowhead key twice to exit out of the configuration menus The value now displayed on the screen is baseline corrected and will continue to be until baseline correction is turned Off A letter C displayed at the end of the 2 display line indicates that baseline correction is currently being applied Note Both the baseline value and the On Off state will be remembered by the instrument across power cycles However selecting the Reset function will turn baseline correction Off and reset the baseline value to zero 28 WORKING WITH THE BE2100 and BE2400 BASE UNITS Analog Output Two analog current outputs are available on the rear panel of the BE2100 Base Unit Both analog outputs produce a standard nominal range of 4 to 20 mA The first analog output AO1 is proportional to the signal being displayed on the front panel of the Base Unit The second analog output AO2 is proportional to rate of change of AOI in log space When AOI is increasing exponentially AO2 thus corresponds to the growth rate of AO in units of inverse hours The B
123. you have not already created these annotations you can do so by hitting the Cancel button right clicking over the graph where you want to start the baseline averaging selecting Create Annotation gt gt Baseline Start End STEP 4 If the Baseline Correction control if set to Off turn it On Press the OK button to put your changes into effect If you want to save your changes to file press the Resave button select a new filename and then press the OK button Note 1 Whenever changes are made the Unsaved Changes button in the Software Settings tab is activated turns from grey to red If you try to exit the program without saving your changes you will first be prompted to make sure you are aware that your changes have not been saved Note 2 The Data Viewer program does not communicate with BE2x00 instruments Changing the baseline setting from within the Data Viewer program affects only the data read from and or saved to file but does not affect the baseline settings stored in sensor memory If you want to change the instrument settings the BE2x00 Virtual Instrument program should be used instead 54 Retrospective Calibration Adjustment The Data Viewer program also provides the capability to retrospectively adjust the user calibration that is applied to the data This feature may be useful if the calibration settings were changed midway through a fermentation run Duri

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