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TD-700 Laboratory Fluorometer Operating Manual

Page 19

3.

Direct Concentration Calibration:  

Direct Concentration Calibration,

only available in the Multi-Optional Mode, is a multi-point calibration in
which up to five standards and a blank are read.  The software uses
these points to set the optimal instrument range and sensitivity, and to
calculate the direct concentration of unknowns. In a multi-point
calibration, the instrument generates a calibration curve for superior
accuracy.  The TD-700 will display the actual concentration of your
samples.  The display units are user-selectable and chosen in the
setup menu.

Note:  Fluorescence measurements are affected by factors such as
temperature, linearity, and instrument drift.  Before calibrating or
running samples for the first time, we recommend reviewing Appendix
1.

Summary of Contents for TD-700

Page 1: ...TD 700 Laboratory Fluorometer Operating Manual Page 1 TD 700 Laboratory Fluorometer Operating Manual Dated 5 8 2002 Version2 0 PartNumber7000 998 ...

Page 2: ...TD 700 Laboratory Fluorometer Operating Manual Page 1 TD 700 Laboratory Fluorometer Operating Manual Dated 5 8 2002 Version2 0 PartNumber7000 998 ...

Page 3: ...uartzHalogenLamp V Instrument Parameters Firmware 13 A Power upScreen B HOMEScreen C Setup CalibrationScreen D Setup Screens E CalibrationScreens F FirmwareFlowChart VI Calibration Overview 18 A WhyCalibrate B WhentoCalibrate C TD 700CalibrationOptions VII Calibration Simple Mode 20 VIII Calibration Multi Optional Mode Raw Fluorescence 22 IX Calibration Multi Optional Mode Direct Concentration 24 ...

Page 4: ...OptionalRawFluorescenceCalibration E Calibration Printout Using the Simple Mode APPENDIX 3 Alarms Diagnostics 36 A AlarmScreen B DiagnosticScreens APPENDIX4 Troubleshooting 38 APPENDIX 5 Data Collection 39 APPENDIX 6 Sample and Cuvette Adapters 40 A InstallingandRemovingSampleAdapters B MinicellAdapter APPENDIX7 Glossary 42 APPENDIX 8 Error Messages and Notes 45 A InvalidInput B Error1 C Notes 1 a...

Page 5: ...TD 700 Laboratory Fluorometer Operating Manual Page 4 NOTES ...

Page 6: ...id located on the top of the unit 3 Remove the Filter Cylinder by grasping the inside rim of the cylinder and pulling up 4 Install the correct optical filters for your application See Section III for details 5 Check that the correct lamp is installed for your primary application See Section IV for details 6 Check to see that the sample adaptor is the correct one for your needs Refer to Appendix 6 ...

Page 7: ...t MSDS forinformationregardinghandling When using potentially hazardous chemicals please use closed cap test tubes whenever possible and limit exposure If closed cap test tubes are not available Parafilm should be used to prevent the spillage of volatile or other potentially hazardous chemicals Chemicals should be used in accordance with local regulations and used in a well ventilated area Do not ...

Page 8: ...y Fluorometer TD 700 Filter Cylinder and 13 mm Sample Adapter 25 mm Round Sample Adapter 10 mm Square Cuvette Adapter II Hardware Overview A TD 700 Quick View Diagram AlignmentMark AlignmentMark SampleAdapter FilterCylinder SilverAlignmentMark SampleChamber Display Keypad ...

Page 9: ...asping the inside rim of the cylinder and pulling it straight up 5 Sample Adapters Adapters are available for various test tube or cuvette sizes The sizes used with the TD 700 are 25 mm round 13 mm round 10 mm square or 100µl minicell 6 Lamp Access Panel To access the lamp remove the panel by pulling on the two black quick release latches on the lamp access panel The lamp provides excitation light...

Page 10: ... want to run Filters are marked on their rim with an identification number c The Filter Cylinder has eight openings for four different filter sets For example the openings for filter set A are marked A EX and A EM filter set B is marked B EX and B EM Select the A B C or D positions to install the filters d Find the openings for the filter set you want to remove For example if you want to install t...

Page 11: ...rored side faces toward the inside of the cylinder toward the sample Insert it mirrored side in if any in the opening marked with the appropriate set letter EM Push the filter in so it rests flush with the back of the opening Then reinsert the o ring and press in with your plastic pen cap until it is flush against thefilter g Position the Filter Cylinder in the sample chamber so that the align men...

Page 12: ...p access panel by pulling on the two black quick release latches on the rear panel of the instru ment 2 Before replacing a lamp make sure the current lamp is fully seated in both the upper and lower lamp sockets Be cautious when remov ing the lamp It may be hot 3 To remove the lamp grasp it carefully and turn it about 90 degrees until the prongs line up with the slots on the lamp sockets Slide it ...

Page 13: ... the instrument The lamp access panel should now pull free of the instru ment 3 DO NOT TOUCH THE LAMP BULB with an ungloved hand the bulb may be hot and or the oils from your hand will affect the lamp s transmittance and cause incorrect readings If the bulb is burned out grab the bulb with a tissue and pull it out of the assembly Using the same tissue insert a new bulb orientation does not matter ...

Page 14: ...complete the countdown period For best results leave the instrument turned on in your laboratory If you wish to bypass the countdown period press H ESC or ENT From the HOME screen press 9 to return to the Power up screen B HOME Screen After the countdown period the HOME screen is displayed The HOME screen is where samples are read and data is sent to the printer or computer Press H to return to th...

Page 15: ...e user through the process Depending upon the calibration mode and procedure chosen in the Setup screens the steps will be different See firmware flowchart on pages 16 17 Here is a description of the 3 differentcalibrationprocedures Simple Onepointcalibration This calibration allows you to use one standard or sample and no blanks The instrument gives the standard a relative value of 500 on a scale...

Page 16: ...TD 700 Laboratory Fluorometer Operating Manual Page 15 NOTES ...

Page 17: ... ENT Setup Cal END XXX units ENT Setup Cal 1 Mode 3 Units 2 Cal Procedure 1 Mode 2 Cal Procedure Simple Mode Direct conc Mode Raw Fluor Mode 1 2 3 K 0 7 8 9 Use the button on the keypad to toggle between options The option in brackets is the selected option 1 Power up Screen Countdown from 600 Home Screen Use this screen to choose between the Simple mode and the other 2 modes Use this screen to ch...

Page 18: ...able XXX Reading Blank Wait Blank XXX Printing Cal Report pls wait Enter highest conc Std first Max conc XXX X New ENT Hi conc XXX X New ENT Number of Stds 1 5 X ENT Max range XXXX 1 OK 9 Change Hi Std Conc XXX X 1 OK 9 Change Insert Hi Std and press K SETTING Sens Send Factor XX Sensitivity Set Sens Factor XX Reading Hi Std WAIT X Std Conc XXX X 1 OK 9 Change X conc XXX X New ENT Insert X Std and...

Page 19: ...omatically set based on the typical standard or sample chosen You cannot manually adjust the instrument range when calibrating in this mode The chosen sample will be automatically set to 50 of the maximum value that can be accurately read by the instrument 500 out of 1000 Also there is no option to read and subtract a blank during this calibration procedure 2 Multi Optional Raw Fluorescence Mode I...

Page 20: ...nstrument range and sensitivity and to calculate the direct concentration of unknowns In a multi point calibration theinstrumentgeneratesacalibrationcurveforsuperior accuracy The TD 700 will display the actual concentration of your samples The display units are user selectable and chosen in the setup menu Note Fluorescence measurements are affected by factors such as temperature linearity and inst...

Page 21: ...nt range and sensitivity Wipe the outside of the test tube or cuvette dry and insert it into the sample adaptor in the sample chamber 4 The TD 700 will now set sensitivity as indicated by the SENS FACTOR sensitivity factor so that the sample you inserted will read 500 half of maximum on the HOME screen 5 Upon finishing the sensitivity adjustment the screen prompts you to acknowledge the value set ...

Page 22: ...TD 700 Laboratory Fluorometer Operating Manual Page 21 NOTES ...

Page 23: ...ed to know the exact concentration you are using it to set the optimal instrument sensitivity and range Wipe the outside of the test tube or cuvette dry and insert it into the sample adaptor in the sample chamber Press ENT to proceed to the next screen 4 Default setting raw fluorescence reading is 800 If this is acceptable press 1 to continue If you want to change this value press 9 Type in the ne...

Page 24: ...y not be able to reach the target sensitivity In these cases an error message will appear indicating that based on the calibration the unit has reached its sensitivity maximum or minimum By pressing ENT you tell the instrument to accept the maximumorminimumsensitivityvalue Itisrecommended however that you adjust your sample concentrations to fall within range and then recalibrate 8 To view the las...

Page 25: ...e press ENT from the HOME Screen press 1 for Setup then 1 again for Mode Use to choose the Multi Optional Mode Press ESC to return to the previous screen then press 2 to choose the calibration procedure Use the key to choose Direct Conc fortheDirectConcentrationcalibrationprocedure Press ESC to return to the previous screen then press 3 to choose the units of measure Use the key to choose the requ...

Page 26: ...for details regarding the calibration data printout B How Sample Concentrations will be Calculated As you have read the Direct Concentration calibration procedure accepts multiple standards and one blank The concentration vs fluorescence plot of both the standard and the blank will result in something close to a straight line assuming the standards are within the linear range of the assay and inst...

Page 27: ... of the line between points 2 and 3 C2 is the concentration of standard 2 C To View the Last Calibration From the HOME Screen press ENT to reach the Setup Cal screen From the Setup Cal screen press D to send the current calibration information to a printer or computer Press ESC to return to the HOMEScreen D To Abort the Calibration To abort the calibration press ESC at any time during the calibrat...

Page 28: ...TD 700 Laboratory Fluorometer Operating Manual Page 27 NOTES ...

Page 29: ...dout The words DLY AVG and END appear on the upper left of the LCD display and alert the user to the instrument s current status After the sequence is complete the display reads END the reading will freeze for 5 seconds and the readout will automatically be sent to your computer or printer The letters disc will appear next to the reading on the printout to indicate that the discrete sample averagi...

Page 30: ...rom the HOME screen 2 Press 9 to abort the procedure or 1 to Reset Blank to Zero Press 1 and without further prompting the instrument will blank to zero 3 If the reading for the blank exceeds the maximum the unit will display an error message Blank level too high If this message should appear evaluate your sample to ensure that you are reading the blank and not another type of sample Once you have...

Page 31: ...itive and even small amounts of material from a previous sample may result in errors Complete rinsing is especially important if you are using the same test tube or cuvette for samples and blank 3 Fill the test tube or cuvette at least 75 full significant error will result if it is not full enough 4 The cuvette MUST BE DRY on the outside when taking readings Moisture on the outside will result in ...

Page 32: ...roportion to the dilution If the reading goes up concentration quenching you are beyond the range for a calibration curve If the reading does not decrease in direct proportion to the dilution you are in the range for a calibration curve but still beyond the linear range D Temperature Considerations Fluorescence is temperature sensitive As the temperature of the sample increases the fluorescence de...

Page 33: ... the test tube that are not visible to the human eye We recom mend that the test tube be marked at the top and positioned in the adaptor the same way each time to minimize error F Data Quality The TD 700 is only as accurate as the standards that are used to calibrate it This is why it is important to take care when preparing standards samples and blanks One should follow good laboratory practices ...

Page 34: ...our computer run the communications program you have chosen For a step by step procedure using Microsoft Windows terminal see Appendix 5 3 Press D or from the HOME screen and the readout currently displayed will be sent to the computer with an index number B Sample Reading with a Printer or Computer When D or is pressed from the HOME Screen the TD 700 sends out an index marker 1 999 the concentrat...

Page 35: ...the photomultiplier voltage Next is the calibration that is stored in the unit s memory Themaximumconcentrationrangeenteredduringthecalibration sequence is displayed here Following this is the Calibration Standard Summary which displays each standard its concentration and its corre spondingrawfluorescencevalue Concentrationandrawfluorescence values are organized in the Calibration Curve Data table...

Page 36: ...the printout display the TD 700 s current parameters the mode the cal procedure the units and the instrument sensitivity level Following this is the calibration stored in the unit s memory The calibration stored consists of the raw and adjusted sample and blank value if appli cable expressed in raw fluorescence units E Calibration Printout Using the Simple Mode Example only Your numbers will vary ...

Page 37: ...ssed from the HOME screen contact Turner Designs If the low power alarm is triggered check the power at the source and the powersupply Refer to the Diagnostic Screens next section for definitions B Diagnostic Screens There are diagnostic screens which show the status of internal fluorometer functions To access these screens from the HOME screen press 8 then ENT to see the next screen in the sequen...

Page 38: ...he unit NOTE To see the raw fluorescence for the standard s and the blank as set during the last calibration from the Calibration Setup menu press D to print or send to a computer NOTE If you calibrated in the Simple Mode or Multi Optional Mode Raw Fluor Calibration Procedure to see the raw fluorescence for the standard and the blank as set during the last calibration from the Calibration Setup me...

Page 39: ...vity Display reads Reading for Blank is less than zero Appendix 8 H negative due to drift reading for samples are less than Blank Reset Blank to Zero orrecalibrate Drifting The instrument is open loop and SectionVII readings readings will drift over a period of time SectionVIII recalibrate to cure SectionIX Erraticreadings Instrumentreadingswillfluctuate SectionIII somewhat due to instrument noise...

Page 40: ... MAC OS use a comparable terminal program to capture the ASCII data 3 Give the New Connection a name such as TD700 4 In the Phone Number window choose Direct to COM 1 or Direct to COM 2 depending on which COM Port that the instrument is connected to This can be the hardest part of the procedure Different computer manufacturers use different conventions 5 Set the Port Settings to the following para...

Page 41: ...able for the TD 700 are a variety of sample adapter to cater to the sample size needs of researchers These are 25 mm test tube adapter P N 7000 982 standard with TD 700 model 7000 000 13 mm test tube adapter P N 7000 981 standard with TD 700 model 7000 000 10x10mmsquarecuvetteadapter PN 7000 988 standard with TD 700 model 7000 009 Minicelladapter P N 7000 951 fits inside 10x10 cuvettes adapter P N...

Page 42: ... Theminicellcuvettesallowsforverysmallvolumes 75 250µl tobe analyzed Toinstall 1 Slide minicell adapter into the 10x10 mm cuvette adapter where a normal cuvette would sit 2 Insert the borosilicate glass minicell into the adapter 3 Insert into the TD 700 as described earlier ...

Page 43: ...rent A small amount of current flows in a photomultiplier tube even when the tube is operated in a completely dark state This output is called dark current and the magni tude is greatly dependent upon the amount of voltage applied to the photomultiplier tube i e the greater the Sensitivity Factor the larger the dark current Detection Limit May be used to refer to either the minimum or maxi mum con...

Page 44: ... The light passes through the sample which emits light proportional to the concentration of the fluorescent material present and proportional to the intensity of the exciting light The emitted light is passed through an emission filter that selects for the appropriate wave length range and the resultant light is then detected by aphotomultipliertube Gain An indicator of the sensitivity of the inst...

Page 45: ...etheTD 700fluorometer ssensitivityona Sensitivity Level scale of 1 to 100 Sens Factor is the term used in the software during calibration Sens can be found on the diagnostic screen and sensitivity level is the term used on the printout The Sensitivity Factor is directly proportionaltothevoltageonthephotomultipliertube The higher the Sensitivity Factor the greater the sensitivity and typically the ...

Page 46: ...itiv ity to the maximum If sensitivity falls below the minimum before reaching the target sensitivity the unit will automatically adjust sensitivity to the minimum Note that this may change the Max range concentration you entered D Error3 The concentration entered for 2 5 Std is greater than the concentration entered for the highest Std Press ENT and enter a lower concentration for the Std OR pres...

Page 47: ...ty calibrate the unit again If you are in the Direct Concentration calibration procedure choose a higher Max range conc or a more concentrated high standard If you are using the Raw Fluor Calibration Procedure use a more concen trated standard and or chose a lower value for the sample setting if you are in the Multi Optional Mode NOTE On rare occasions you might see OVER This means that the raw fl...

Page 48: ...esigns warrants the TD 700 Fluorometer and accessories to be free from defects in materials and workmanship under normal use and service for a period of one year from the time of initial purchase with the followingrestrictions 1 The instrument and accessories must be installed powered and operated in compliance with the directions in this TD 700 Laboratory FluorometerOperatingManualanddirectionsac...

Page 49: ...ributor If you have purchased direct contact us We will repair the instrument at no charge but we will not pay for shipment documentation etc These charges will be billed at cost NOTE Under no conditions should the instrument or accessories be returned without notice Prior correspondence is needed a To ensure that the problem is not a trivial one easily handled in yourlaboratory withconsequentsavi...

Page 50: ...3 86 x 86 9 8 cm x 2 18 cm Keypad 4 x 5 keys 3 x 2 7 7 62 cm x 6 86 cm DataOutput 100 ASCIIformatthrougha9 pinRS 232 serial cable at 9600 baud Filter Cylinder Accommodateseight 1 inchroundfilters four excitation and four emission Software Menu drivenmicroprocessor controlled Detector Factory installedphotomultipliertube Stan dard 300 650 nm Optional Red Sensitive 185 870 nm Lamp Quartz Halogen Lam...

Page 51: ...librationforDirectConcentration Measurement or Single Point Raw Fluores cenceCalibration Operating Mode Simple for the inexperienced user or Multi Optional fortheadvanceduser Blank Reads and subtracts Blank Discrete Sample Averaging Samplereadingscanbeaveragedtoimprove accuracy 7 second delay 12 second signal averaging 5 seconddisplayreadout Warranty One yearwarranty Approvals CE TUV and UL ...

Page 52: ...o their ground state These excitation and emission wavelengths or spectra are often referred to as the compound s fluorescence signature Figure 2 shows the key components of a filterfluorometer Figure 2 Principles of Filter Fluorometry Lamp The lamp or light source provides the light energy that excites the compound of interest The lamp actually provides a broader range of light than that which ex...

Page 53: ...solution at a right angle to the incident light energy The size of the sample cell will also effect your measurement The greater the diameter of the cuvette the lower the concentration that can be read Emission Filter Stray light and any background material fluorescence will also be emitted from the sample solution The emission filter is chosen to screen out these components allowing only waveleng...

Page 54: ... located at the bottom of the standard as the high concentration H 7 Removesolidstandardandreplaceanycuvetteadaptorsthatwereremoved in step 2 8 Record instrument settings on TD 700 Calibration Record with Solid Standard sheet that has been included for future reference 9 Proceed with analyzing your samples 10 You may use your solid standard at any time to check the stability of the instrument Simp...

Page 55: ...oratory Fluorometer Operating Manual Page 54 TD 700 Calibration Record with Solid Standard Instr Calibration Mode CalProcedure Maximum L H Date S N Std Conc s Units SimpleorMulti RaworDirect Range Reading Reading ...

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