Fluorimeter Manual
Contents
1. Use the up and down arrow keys to highlight a menu option press the enter key to confirm EXIT METHOD ID B HAME GUIHIHE f EXCITATION nm 350 EMISSION nm s550 UNITS PER RESOLUTION G El CALIBRATION SETUP EXIT LOCKED orf LOCKING CODE ait LAHGUAGE English TIME 13 38 86 DATE 19 84 66 DATE FORMAT DDAA OPERATOR ID S06 keys press the enter key to confirm the setting when ALL digits have been correctly set Use the up or down arrow keys to browse through pre set options when the correct selection Left arrow eiT Up arrow ke LOCKED OFF LOCEING CODE aed LANGUAGE English Right arrow TIME 13 42 82 DATE 19704706 Down arrow DATE FORMAT DEMM YY Down arrow OPERATOR ID 200 is displayed press the enter key to confirm your choice 8 Section 3 Fluorescence Theory The interaction between electromagnetic radiation and matter provides a useful qualitative and quantitative analytical tool known as spectroscopy The region of the electromagnetic spectrum to which matter under investigation is subjected to defines the type of transitions that occur within the molecules Fluorimetry uses radiation from the UV Vis region of the electromagnetic spectrum to study transitions between electronic levels in a molecule or atom The absorption of energy from light radiation photons by a molecule or atom promotes electrons from a low energy ground2. If the concentration and other data from the calibration are correct and satisfactory select ACCEPT to continue through the calibration sequence to the first standard If the previous step has given unacceptable figures then select REPEAT and perform the previous step again ABORT allows the whole calibration sequence to be cancelled and the instrument to return to the measure screen Once the blank calibration has been accepted the instrument will display the message Please insert standard x xxx and the concentration of the first standard as set in the STANDARDS TABLE of the MEASURE CALIBRATION SETUP Chlororhell wos Please insert standard 16 66 ABORT CAL Remove the cuvette of Blank solution from the sample chamber replace with a cuvette of the requested standard and close the lid If using reusable cuvettes ensure that proper cleaning procedures are employed to make certain that there is no carry over of solutions or contamination As with the blank calibrate the instrument with this first standard by using the CAL option on screen or CAL key As with the blank the concentration of the standard as set in the STANDARDS TABLE will be shown on screen with the raw fluorescence and the percentages of full scale emission for the signal and reference Once again review this data and if acceptable use the ACCEPT option to move on to any further calibration standards in the same way If not the REPEAT function can be
3. STATISTICS screen To move to STATISTICS use the option at the bottom of the GRAPH screen STATISTICS SLOPE H 813 k SQUARED 6 922 8 The STATISTICS screen shows two numbers calculated from the saved results The SLOPE reveals the extent of change of fluorescence over time The R squared value gives an indication of the linearity of the saved results Section 12 Printing results Printing 1 When a reading has been taken in the MEASURE mode the concentration raw fluorescence result is displayed on screen This can be printed to a serial printer or PC if attached via the RS232 The fluorimeter is designed for use with the optional accessories of the Porti printer P N 037 701 or DataWay PC software P N 050 501 Porti printer 1 Connect the Porti printer to the RS232 socket on the back panel of the instrument using the supplied cable 2 The printer can be used without connection to a mains power supply for periods of time by means of the integral battery If the battery is low or mains power is preferred plug in the power supply pack using the appropriate cables as provided with the printer 3 Ensure the printer is fitted with a roll of thermal paper Paper can be advanced by use of the Feed button on the printer 4 Switch on the printer with the black button and the left hand side LED will light The LED will be green when mains power is used or the battery is well charged If the LED shows flashes of
4. filter is missing the instrument will not allow measurement to take place Pressing the EXIT key will return the screen to the setup menu options UNITS allows user to select the units appropriate to the test ppm RFU U ml mU I U I uM A mM M I ug ml mg ml ug di mg dl g dl ng l ug l mg l g l none ppb RESOLUTION 0 01 0 1 1 0 001 CALIBRATION SETUP this menu is not displayed when RFU Raw Fluorescence units are selected EIT HO OF STOS 3 STAHDARDS TABLE FLUOQRESCEMCE TABLE Pit T GAIHi xa B46 AUTO SET GAIH CAL REMINDER OFF CAL IHTERUALCHRSI Gl EXIT allows the user to return to the previous menu NO OF STDS allows selection of 1 6 standards blank is always included STANDARDS TABLE user entered calibration standards 11 FLUORESCENCE TABLE shows results of last calibration RFU raw fluorescence values Tabular and graphical displays available FLUORESCENCE TABLE _ Fi 2 385 F2 41 229 F3 118 316 n CSTO 100 EXIT WIE CURUE EXIT P M T GAIN 0 to 100 gain setting for system photomultiplier tube Note This option is not available on the Model 6270 AUTO SET GAIN uses the highest user entered calibration standard to automatically set the PMT gain to give as close as possible to 95 full scale emission signal Note This option is not available on the Model 6270 CAL REMINDER enables or disables the measure mode prompt to
5. fluorimeter is powered by a low voltage dc power supply that operates from a 90 264Vac mains supply The universal power supply is supplied with 3 mains leads for UK EU and US sockets The correct lead for your supply should be selected 2 23 Mains Connections The leads supplied have a moulded on plug However if this is removed for any reason the wires in the mains lead are colour coded to conform to the internationally recognised standard such that UK CONNECTIONS US CONNECTIONS BROWN LIVE BLACK LIVE BLUE NEUTRAL WHITE NEUTRAL GREEN YELLOW EARTH GREEN EARTH Safety When disposing of any removed plug the connectors must be removed or made incapable of insertion into a mains socket 4 2 24 Keypad Functions d 1 UP ARROW key used to navigate through menus to increase values and for paging up in stored results 2 DOWN ARROW key used to navigate through menus to decrease values and for paging down in stored results 3 LEFT ARROW key used to navigate through menus and to highlight the selected digit when setting values 4 RIGHT ARROW key used to navigate through menus and to highlight the selected digit when setting values 5 ENTER key used to accept the highlighted menu option 6 CAL key initiates a calibration sequence from within the measurement mode 7 PRINT key initiates a print from the measurement display or stored results Sends data to RS232 serial port 2 25 Instrumen
6. locked without the entry of a locking code EXIT LOCKED Dnj LOCKIHG CODE Ai LANGUAGE English TIME 13 36 54 DATE 19 84 86 DATE FORMAT DOMM YY OPERATOR ID eae 14 5 To set or change the locking code press the down arrow key to select LOCKING CODE then press the enter key EXIT LOCKED On LOCEIHG CODE Els 1 LAMGUAGE English TINE 14 47 ag DATE 19 64 66 DATE FORMAT DDS MAS YY OPERATOR ID 506 6 Adjust the highlighted digit using the up or down arrow keys then select the other digits using the right or left arrow keys Adjust each in turn until the code of your choice is displayed Press the enter key to confirm this ensuring that you can remember the number selected 7 All methods are now locked against adjustment to the set parameters and a padlock icon will be displayed at the top of the method settings to indicate this It is not possible to move between methods 8 Access to the locking code from the instrument setup menu will now require the entry of the locking code first 9 To exit the INSTRUMENT SETUP menu use the up or down arrow keys to highlight EXIT then press the enter key To set additional parameters continue as detailed in the following text EXIT El METHOD ID BS HAME Chlororhsll pee id EXCITATION nm 438 as EMISSION nm 668 UNITS mail ERO RESOLUTION 8 81 CALIBRATION SETUP Language 1 Press the down arrow key to move the highlight to LANGUAGE a
7. s blank solution and standard solutions as set in the CALIBRATION SETUP of the MEASURE SETUP menu Using the steps as detailed in the Using a stored method section select the required method Once in measure mode press the CAL key once to initiate the calibration sequence The instrument display will show the message Please insert blank Chlororrrsl1 marl Please insert blank ABORT CAL Taking care not to mark the optical surfaces place the cuvette containing the blank solution in the cuvette holder of the sample chamber and close the lid Measure the fluorescence of the blank by pressing either the CAL key or with the CAL option highlighted on the instrument screen Press the enter key to confirm The instrument screen will show the status message Reading for the duration of the measurement Chlororrrs1l1 marl Reading Please insert blank ABORT CAL Once complete the concentration zero as this is the blank will be shown as the large number on screen Below this are the raw fluorescence and the percentages of full scale emission for the signal and reference Chlororrrs l1 0 00 8 81 RFU SIG B 112 F REF r6 ro4 APS ABORT REPEAT 22 9 10 11 12 13 14 15 16 17 The options at the bottom of the screen will have also changed The ACCEPT and REPEAT options relate to the calibration step just completed
8. used Chlarorhull 10 00 d d RFU SIG 4 645 XF3 REF f u rrd APS ABORT REPEAT The sequence of calibrating with each standard will continue in this fashion points 10 13 until each standard that has been set has been used At this point the instrument will give a summary of the calibration by displaying the raw fluorescence for each standard in a table An option to view the calibration curve a plot of the concentration of the standards against their raw fluorescence is given at the bottom of the screen VIEW CURVE On exiting the calibration summary the instrument is returned to the measure screen and the status message Cal required will have been removed Measurements of unknown concentration samples can now be made 23 Section 9 9 Performing measurements Using the steps as detailed in the Using a stored method to select the required method Prior to taking any measurements make certain that the correct excitation or emission filter modules are fitted and that a valid calibration as detailed in Section 8 has been completed Before taking any measurements it is worth considering the utilisation of functions such as AUTO SAVE and AUTO PRINT to provide easy access and analysis of results at a later time These are fully explained in the following section Saving results and Printing results If readings are required at regular timed intervals the TIMED READ facility can be applied
9. 40nm 627 139 Wavelength 450nm 31 Interference Filters continued 627 167 Wavelength 460nm 627 136 Wavelength 470nm 627 168 Wavelength 480nm 627 169 Wavelength 490nm 627 170 Wavelength 500nm 627 171 Wavelength 510nm 627 137 Wavelength 515nm 627 172 Wavelength 520nm 627 173 Wavelength 530nm 627 174 Wavelength 532nm 627 175 Wavelength 540nm 627 176 Wavelength 546nm 627 177 Wavelength 550nm 627 178 Wavelength 560nm 627 179 Wavelength 570nm 627 180 Wavelength 577nm 627 181 Wavelength 580nm 627 182 Wavelength 590nm 627 183 Wavelength 600nm 627 184 Wavelength 610nm 627 185 Wavelength 620nm 627 186 Wavelength 630nm 627 187 Wavelength 633nm 627 188 Wavelength 640nm 627 189 Wavelength 650nm 627 190 Wavelength 656nm 627 191 Wavelength 660nm 627 192 Wavelength 670nm 627 193 Wavelength 680nm 627 194 Wavelength 690nm 627 195 Wavelength 694nm 627 196 Wavelength 700nm Alternative filters available on request Sample Cells A wide range of cells in addition to those listed is available to special order Details supplied on request 035 120 100ul quartz cell 035 121 500ul glass cell 035 122 500ul quartz cell 060 247 Pack 100 10mm plastic 3ml cuvettes 060 253 10x10mm glass cell UV 060 254 10x10mm glass cell Visible 060 255 Flow cell Additional Items 628 200 Heated cell system comprising heated cell holder controller 2 leads 021 057 12V connection lead 630 028 Dust Cover 050 501 DataWay PC Software CD ROM wit
10. Fluorimeters Models 6270 6280 amp 6285 Operating Manual 628 020 REV A 04 06 Safety This is important information please read carefully before installing or using this instrument 1 The 62 Series Fluorimeters are designed for operation by trained personnel that are aware of the principles and applications involved For further help and advice please contact your local distributor e mail sales jenway com or visit www jenway com The fluorimeter is a sensitive electronic and optical instrument designed for use in a laboratory environment Careful adherence to the installation instructions must be observed If in doubt contact a relevant and competent authority for advice before proceeding In addition to observing the instructions detailed in the Operating Manual and Service Manual for this instrument all installation operating and service personnel must be aware of and employ a safe system of work Voltage levels hazardous to life are present in this instrument for personal safety only trained engineers aware of the risk and avoidance of electric shock should remove protective covers from the instrument This instrument is designed for minimal maintenance which must be carried out carefully following the procedures detailed in this manual All safety instructions in these procedures as well as those defined locally for the area or environment where the work is being carried out must be observed Other than for th
11. IT using the right and left arrow keys and press enter to confirm This will return you to the main menu options Measure Setup menu Enter MEASURE SETUP menu by selecting SETUP from the MAIN menu The following screen will be shown EXIT METHOD ID BS HAME Chlororhsl1 EXCITATION nm 438 EMISSION rnm 668 UHITS mail RESOLUTION Bal CALIBRATION SETUP EXIT allows user to return to the previous menu METHOD ID used to index individual methods 0 to 19 10 NAME user entered method name up to a maximum of 19 characters Chlororh EXCITATION NM user set excitation wavelengths Actual filter data is checked against this setting during measurement If an incorrect filter is detected a warning will be displayed EMISSION NM user set emission wavelengths Actual filter data is checked against this setting during measurement If an incorrect filter is detected a warning will be displayed Note If the excitation or emission filter modules do not match the current method settings on entry to the measure mode or prior to taking a reading or calibration then a warning is displayed WARH IHG IHCORRECT ERCITATIOH FILTER FITTED EXCITATION rnm 358 METHOD SETTIHG 436 FILTER TYPE BP PART HO 627 126 ASE CHANGE METHOD SE EXIT A scrolling message will appear please change method setting or fit correct filter module No measurements can be performed until this has been corrected If a
12. The use and setting of this is detailed separately in section Timed readings below Once in measure mode place the sample solution of unknown concentration to be measured into the cuvette holder of the sample chamber and close the lid Move the on screen highlight using the left and right arrow keys to the READ option and press the enter key A status message Reading will be shown throughout the duration of the measurement Once the measurement is complete the calculated concentration of the sample will be shown at the resolution and with the units of measure as set in MEASURE SETUP This measurement can then be saved or printed as detailed in the sections below entitled Saving results and Printing results 10 Further measurements can be taken by use of the READ option NOTE AUTO SAVE AUTO PRINT and TIMED READ are all related to the selected method 24 Section 10 EON NOON 10 Timed readings To ensure that all measurements are taken at the required interval select the required method as described in section Using a stored method above then return to the main menu Enter the SETUP menu and select the RESULTS STORAGE section Moving down to the first parameter TIMED READ set this to On The interval between the end of one read and the beginning of the next read can then be set by selection of INTERVAL Using the up and down arrow keys set the numerical figure to the required period i
13. ase consult the DataWay help file Auto print To remove the need to manually select PRINT after each result AUTO PRINT can be activated After every read operation either timed or manual the concentration will be automatically printed Select the required accessory to which to print results to and setup as detailed above in either the Porti printer or Datavvay section To ensure every result is printed select the required method as detailed in Using a stored method and then return to the main menu Enter the setup menu and select the results storage section Move to AUTO PRINT and set to On Using the EXIT options return to the main menu and then move to the MEASURE screen Each read operation will now instigate the result being automatically printed to the connected device 29 Section 13 Maintenance amp Troubleshooting 8 1 General The 62 Series Fluorimeters are designed to give optimum performance with minimum maintenance It is only necessary to keep the external surfaces clean and free from dust The sample area should always be kept clean and any accidental spillage should be wiped away immediately To give added protection when not in use the unit should be disconnected from the mains supply and covered with the optional dust cover 630 028 For longer term storage or re shipment it is recommended that the unit be returned to the original packing case 8 2 Xenon Lamp Replacement In the highly unlik
14. calibrate at the set calibration interval CAL INTERVAL HRS the number of hours between calibrations can be user set from 1 999 hours Results Storage Menu From the MAIN MENU press the down arrow key to move the highlight on the display to cover RESULTS STORAGE then press the enter key The following display will be shown EIT TIMED FREAD Or IHTERUAL ts BES AUTO SAVE ort AUTO PRIHT orf EXIT allows the user to return to the previous menu TIMED READ enables automatic measurements to be taken at a set interval when in measure mod INTERVAL time interval between the end of one reading and the automatic start of the next reading in seconds AUTO SAVE when enabled automatically saves to instrument memory after each reading is performed either manually or timed AUTO PRINT when enabled automatically prints to RS232 connected device after each reading is performed either manually or timed 12 Instrument Setup Menu From the MAIN MENU press the down arrow key to move the highlight on the display to cover INSTRUMENT SETUP then press the enter key The following display will be shown EIT LOCKED ort LOCK IMG CODE Hal LAMGUAGE English TIME 21 59 47 DATE 83 82 68 DATE FORMAT DD OPERATOR ID Hpg EXIT allows the user to return to the previous menu LOCKED when enabled prevents access to set up menus without the PIN code settings of current method to be viewed but not changed Calib
15. data will be displayed here From the FLUORESCENCE TABLE it is also possible to view the calibration curve by choosing the VIEW CURVE option As with the data table this will show default values until a valid calibration has been completed 19 FLUORESCEHCE TABLE Fi 2 355 Fi 41 229 F3 118 716 OO SEE 0 CSTD 100 Es IT WIEW CURVE EXIT 13 Set the photomultiplier tube gain P M T GAIN to the required percentage for this test if known If not known use the AUTO SET GAIN option as below Note This option is not available on the Model 6270 14 To find the optimum gain for a given test select the AUTO SET GAIN menu option and follow the instructions on the instrument screen If you accept the gain suggested by the instrument this will then automatically update the gain setting in the CALIBRATION SETUP menu Note This option is not available on the Model 6270 AUTO SET GAIH AUTO SET GAIH AUTO SET GAIH MEASURING mx Full Scale Signal PRERE MO CET Full Scale Signal 89 ee B 834 hae 38 88 STAMDARD t Cain x Gain 20 66 ABORT REAL ABORT READ ABORT REFEAT 15 To ensure that calibrations are regularly performed the final two CALIBRATION SETUP menu options can be employed If you wish for a calibration reminder to appear in the measure screen at regular intervals set the CAL REMINDER to On The interval between calibration and the reminder being shown is set using CAL INTERVAL to th
16. e The clock can only be set in the 24 hour system i e 1 00 p m 13 00 00 4 To exit the INSTRUMENT SETUP use the up or down arrow keys to highlight EXIT the press the enter key To set additional parameters continue as detailed in the following text Date EXIT EXIT EXIT LOCKED OFF LOCKED OFF LOCKED OFF LOCEIHG CODE Bed LOCKING CODE Bed LOCEIHG CODE dni LAHGUAGE English LANGUAGE English LANGUAGE English TIME 14547588 TIME 14 45 81 TIHE 14 45 81 DATE 19 84 86 DATE 19 64 66 DATE 19 646 DATE FORMAT MER LA LS DATE FORMAT EEES DATE FORMAT GGMEEEETS OPERATOR ID pe OPERATOR ID SHH OPERATOR ID pe 1 Press the down arrow key to highlight DATE then press the enter key 2 Use the up and down arrow keys to set the highlighted digit of the date to the correct value Use the right and left arrow keys to highlight each digit in turn for adjustment with the up and down arrow keys until the correct date is set 3 Press the enter key to confirm the setting and start the date running from the set value 16 4 Use the down arrow key to select DATE FORMAT Press the up or down arrow keys to view the two alternative date formats Press the enter key to confirm your choice The previously set date will automatically change with your selection in this menu option 5 To exit the INSTRUMENT SETUP use the up or down arrow keys to highlight EXIT then press the enter key To set additional parameters continue as detailed in the following text Op
17. e required number of hours using the up and down arrow keys EIT EIT HO OF STDS 3 HO OF STOS 3 STANGARDS TABLE STAHDARDS TABLE FLUORESCENCE TABLE FLUORESCEMCE TABLE P M 7T GAIHEA B48 F H T GAIHC I Ad AUTO SET GAIH AUTO SET GAIH CAL REMINDER Dr CAL REHIHDCER orf CAL INTERWALCHRS 1 CAL INTERWALCHES gt dE 20 16 The method is now fully set and will be the available in the MEASURE screen EXIT HO OF STOS 3 STAHDARDS TABLE FLUORESCEHCE TABLE P M T GAIHEX B66 AUTO SET GAIN CAL REMINDER On CAL INTERWALCHRS AZ Section 7 Using a stored method 1 From the MAIN menu select the SETUP menu and then MEASURE SETUP 2 By setting the METHOD ID to the appropriate number 0 19 the saved parameters such as the method name and wavelengths will appear and be ready for use 3 EXIT the setup menus and return to the MAIN menu 4 Ensure that the excitation and emission filters appropriate to selected method are fitted in the sample chamber 5 Enter the MEASURE mode 6 The name of the selected method will be shown at the top of the screen with the PMT gain as set for that method also displayed Note This option is not available on the Model 6270 7 No concentration will be shown as no measurement has been taken but the selected units of measure will be displayed 21 Section 8 Calibration Before beginning the calibration ensure that close to hand are the appropriate cuvette
18. ecified limits before carrying out the analysis Section 2 Getting Started 2 1 Unpacking Remove the universal 12V power supply with UK US and EU leads and the pack of 100 disposable cuvettes from the packaging Remove the fluorimeter from the carton by lifting it in the centre between the two support cheeks do not lift it by the support cheeks Place all items on a clean workbench then remove the support cheeks and the polythene bag from the fluorimeter Any shortages or damage must be reported to your local distributor or the manufacturer as soon as possible Keep all packing materials in case the unit has to be re shipped at a later date It is important that when re packing the instrument it is first sealed in a strong clean polythene bag to protect it from the dust and particles that are present in all packing materials 2 2 Installation 2 21 Location In ideal circumstances the installation environment will be clean dry and dust free with the instrument protected from extreme variations in ambient lighting and temperature change For field use it is recommended that the instrument is used in the optional storage case for additional protection Where conditions are less than ideal maintenance and cleaning must be carried out regularly and additional protection offered where possible The optional dust cover should always be fitted when the unit is not being used or is stored for short periods 2 22 Supply Voltage The
19. ely event of lamp failure it will be necessary to contact your local distributor or service agent for assistance There are no serviceable parts within this product Under no circumstances should any attempt be made to effect repair as this will invalidate the product Warranty 30 Section 14 Optional Accessories 14 1 Accessories Bandpass Filters 627 126 UG1 320 380nm 627 124 BG28 380 500nm 627 125 VG9 480 580nm Cut off Filters low cut transmission blocked below stated wavelength 627 127 Kodak 29 610nm 627 128 Ilford 201 545nm 627 129 Kodak 8 475nm 627 130 Kodak 2B 395nm 627 131 Glass 305nm Interference Filters 10nm bandpass 627 141 Wavelength 250nm 627 132 Wavelength 254nm 627 142 Wavelength 260nm 627 143 Wavelength 265nm 627 144 Wavelength 270nm 627 145 Wavelength 280nm 627 146 Wavelength 290nm 627 147 Wavelength 295nm 627 148 Wavelength 300nm 627 149 Wavelength 305nm 627 150 Wavelength 310nm 627 151 Wavelength 313nm 627 152 Wavelength 320nm 627 153 Wavelength 326nm 627 154 Wavelength 330nm 627 155 Wavelength 334nm 627 156 Wavelength 337nm 627 133 Wavelength 340nm 627 140 Wavelength 350nm 627 157 Wavelength 360nm 627 138 Wavelength 365nm 627 158 Wavelength 370nm 627 134 Wavelength 380nm 627 159 Wavelength 390nm 627 135 Wavelength 400nm 627 160 Wavelength 405nm 627 161 Wavelength 410nm 627 162 Wavelength 415nm 627 163 Wavelength 420nm 627 164 Wavelength 430nm 627 165 Wavelength 436nm 627 166 Wavelength 4
20. entered to support conformance and traceability of operation With press to read operation and Total energy transfer TET technology the output of the high energy xenon lamp is maximised and its expected life extended so that it should never need replacing in normal use The high quality optics is complemented by the Intelligent Filter Modules IFM that can be identified by the system this enables error messages to be generated and displayed should the wrong filters be fitted or their positions be incorrect All models are powered from an external universal mains adapter suitable for use from 90Vac to 264Vac while the 12V dc input enables use of the fluorimeters in vehicles or from suitable battery packs Where temperature controlled fluorescence studies are necessary an optional electrically heated sample holder is easily fitted while for continuous flow analysis an external sipper pump and a wide range of flow through cuvettes are also available 1 2 Good Practice Guidelines 1 The selection of the optimum excitation and emission wavelengths filters is critical to achieving the best performance from the analysis 2 All fluorimeters should be sited in a clean dry dust free environment When in use ambient temperature and light levels should remain as constant as possible 3 Adherance to Standard Operating Procedures SOP and Good Laboratory Practice GLP should be maintained with regular calibration checks and a suitable Qual
21. erator ID OPERATOR ID is a 3 digit code that can be quickly and easily entered from the INSTRUMENT SETUP menu This ID will appear in the header of all results printed or down loaded from the fluorimeter identifying the operator that carried out the analysis When using this facility all potential users should be allocated individual 3 digit codes Operators should enter their code using the following procedure 1 Use the up or down arrow keys to select OPERATOR ID from the INSTRUMENT SETUP menu then press the enter key EIT EIT LOCKED DFF LOCKED DFF LOCEIHG CODE dni LOCEIHG CODE Bal LANGUAGE English LANGUAGE English TIME 14 56 57 TIME 14 58 42 DATE B4 19 86 DATE 4 19 66 DATE FORMAT MH DD YY DATE FORMAT HH DD Sha OPERATOR ID SH 2 Use the right or left arrow keys to highlight each digit in turn Use the up or down arrow keys for adjustment until the new operator ID is set 3 Confirm the operator ID by pressing the enter key This current ID will be used in the header of all result printouts and results data transferred to a PC or other serial device 4 To exit the INSTRUMENT SETUP menu use the up or down arrow keys to highlight EXIT then press the enter key This will return you to the main menu 17 Section 6 Setting up a method 1 Enter MEASURE SETUP menu by selecting SETUP from the MAIN menu 2 Select a suitable METHOD ID 0 19 3 Give the method a NAME up to 19 characters using the alp
22. h interface cables and instructions 033 290 Carry Case 021 056 Universal 12V power supply with UK EU and US mains leads 32 Section 15 Specification amp Data 10 1 Technical Specification Specification 6270 6280 6285 Light source Pulsed Xenon Lamp with press to read operation Expected Lifetime No replacement required in normal operation Sensitivity Quinine Sulphate 1pg ml lt 1ng ml lt 1ng ml Dynamic Range Quinine 5 2x 10 5 2 x 10 5 2x 10 Sulphate Detector Photodiode Photomultiplier Photomultiplier Wavelength Range 190 to 1100nm 190 to 650nm 190 to 850nm Wavelength Selection Intelligent Filter Modules IFM Raw Fluorescence v v v Calibration Blank Blank Blank Concentration v v v Calibration Blank up to 6 Blank Up to 6 Blank up to 6 standards standards standards Kinetics Timed Readings In both above In both above In both above modes modes modes Time Limits 1 reading every 3 to 999 seconds Methods Stored 20 20 20 Results Stored 100 method 100 method 100 method GLP Method lock with password access Time and date stamped results Last calibration date and time stored Calibration Interval set Calibration due reminder User ID Outputs RS232 Serial Port PC Software Compatible with DataWay option Sample Handling 10mm Square cuvette holder standard Heated Sample Holder Option Sipper Pump Option Powe
23. hanumeric screen accessed by selecting the NAME option of the MEASURE SETUP menu EXIT Chlororh EXIT METHOD ID HS METHOD ID AS MAME Chlororhull MAME Chlororhull EXCITATION rnm 4348 EXCITATION nm 4348 EHISSIOH nm 668 EMISSION nm 668 UNITS marl UHITS marl RESOLUTIOH H i RESOLUTION Al CALIBRATION SETUP CALIBRATION SETUP 4 Return to the MEASURE SETUP menu and set the EXCITATION wavelength in nm for the test you wish to perform The excitation filter module fitted for this test will need to be the same wavelength 5 Inthe same way set the EMISSION filter wavelength ensuring this is the same wavelength as the emission filter module 6 By scrolling through the available units select the required UNITS OF MEASURE EXIT EXIT EXIT METHOD ID GS METHOD ID aS METHOD ID ss HAME Chlororhs11 HAME Chlororhull HAME Chlororhs11 EXCITATION rim 436 EXCITATION nm 434 EXCITATION rm 438 EMISSION nm 668 SSID 668 EMISSION nm 668 UHITS mal UNITS marl UHITS ma Ll RESOLUTIOH 61 RESOLUTION A Bl RESOLUTION 6 81 CALIBRATION SETUP CALIBRATION SETUP CALIBRATION SETUP 7 If you wish to make raw fluorescence measurements then select RFU from the units of measure If RFU is used then there is no need to calibrate before taking measurements so the CALIBRATION SETUP menu at the bottom of the MEASURE SETUP menu is removed EXIT METHOD ID Be HAME Chlororhs11 ESCITATIOH rnm 436 EMISSION r
24. he method currently selected in MEASURE SETUP return to the MAIN menu Using the down arrow key move the highlight to RESULTS and press the enter key A table of all saved results for the method named at the top of the screen will be shown along with the time at which the READ operation occurred The results are presented in chronological order beginning with the most recent If the number of stored results exceeds seven a small down arrow will be shown on the right hand side of the screen to indicate further screens of results These other results are accessed by use of the down arrow key The full range of results can be viewed by use of the up and down arrow keys to move to the screen of interest Chlororhy1l1 72 83 15 21 01 3 H 15 20 59 3 24 15 28 55 3 28 15 28 58 3 69 15 28 42 3 87 15 26 29 3 28 15 20 75 EXIT GRAPH CLEAR The Clear function on this screen will remove ALL stored results for the Method named at the top of the screen Once Clear has been utilised the results cannot be recovered A graphical representation of the stored results plotted against time is available This is accessed by the GRAPH option at the bottom of the results table screen 0 SECS 10 EXIT STATISTICS The first earliest result is set as time zero and the times of subsequent results are calculated from this but purely for the graph 27 7 Further analysis of the saved results can be made by accessing the
25. ity Control QC programme 4 The correct selection of cuvettes is imperative for accurate and reproducible results a Ensure all cuvettes used are compatible with fluorimetric measurements where the emission beam is at 90 to the excitation source Typical examples have 4 clear sides b Ensure any native fluorescence from the cuvette material is minimal at the analysis wavelengths c Plastic cuvettes should be used once only d Glass and quartz cuvettes should be thoroughly cleaned after use Discard when scratches or marks are evident in their optical surfaces e Ensure any cleaning agents used do not fluoresce at the analytical wavelengths and are thoroughly rinsed away before drying f Ensure the cuvettes used are compatible with the constituents of both the samples and standards they are to hold Plastic cuvettes are not compatible with some organic solvents g Cuvettes must be handled with care by the top and non optical surfaces only Any finger marks must be removed by using a suitable cleaning process h Flow through cuvettes must be selected with additional consideration for the sample type aspirated volume pumping system and rinse cycle as well as the waste handling to be used 5 The high sensitivity of fluorimetric analysis means that all glassware used in the preparation of samples and standards must be totally free from contamination 6 Chemicals and reagents used in sample and standard preparation should be
26. l pass all wavelengths above their designated cut off point this makes them ideal as an emission filter blocking any stray radiation from the shorter wavelength excitation source but beware of scatter which will be at longer wavelengths and so will be transmitted with the fluorescence Interference filters are available across the full wavelength range and are specified by their peak wavelength and spectral bandwidth SBW With typical SBWs of 10nm their performance is close to that of a monochromator and all unwanted wavelengths can be excluded TET Total Energy Transfer IFM Intelligent Filter Modules Filters are pre programmed with their peak wavelength and type These are identified by the system enabling error messages to be generated and displayed should the wrong filters be fitted or their positions be incorrect 34 EC Declaration of Conformity Jenway Models 6270 6280 and 6285 Fluorimeters comply with the following European Standards EN 50081 1 1992 Electromagnetic compatibility Generic emission standard EN 50082 1 1992 Electromagnetic compatibility Generic immunity standard EN 61010 1 2001 Safety requirements for electrical equipment for measurement control and laboratory use Following the provision of EMC Directive 89 336 EEC and Low Voltage Directive 73 23 EEC cum Carl Warren General Manager Jenway Gransmore Green Felsted Dunmow Essex CM6 3LB England 35
27. n seconds between readings EXIT TIMED READ On INTERWAL s3 B36 AUTO SAVE On AUTO FRIHT ort The instrument can now be returned to the MEASURE mode Place the cuvette of sample in the sample chamber and close the lid To begin taking timed measurements highlight READ and then press the enter key At this point the first measurement will be taken indicated by the status message Reading Once this first read is complete a further message will be shown which displays a countdown in seconds to the next read When this timer reaches zero the next reading will be taken Chlororhy1l1 0 28 Fesult Saved Timed Read In GZ3 SYSTEM GAIH 66 EXIT SALE It is especially important when interval times used are short to use either the AUTO SAVE or AUTO PRINT in conjunction with timed readings This enables full automation of result collection without the user having to manually record readings for the entire duration of the testing To cease the timer readings move the highlight to EXIT and press the enter key This will return the instrument to the main menu NOTE AUTO SAVE AUTO PRINT and TIMED READ are all related to the selected method 25 Section 11 Saving results Saving results When a reading has been taken in the MEASURE mode the concentration result is displayed on screen and can be saved to the instrument internal memory by selecting the SAVE option on the right hand side
28. nd then press the enter key EIT EIT LOCKED DFF LOCKED off LOCEIHG CODE dni LOCEIHG CODE Bal LANGUAGE English LANGUAGE English TIME 14 38 55 TIHE 14 27 59 DATE 19 64 86 DATE 19 846 DATE FORMAT DOMM YY DATE FORMAT DOMM YY OPERATOR ID a06 OPERATOR ID Sha 2 Use the up or down arrow keys to view the alternative languages available English Spanish Italian German or French 15 3 When the language of your choice is highlighted press the enter key to confirm The language used on the display will now be changed to match your choice 4 To exit the INSTRUMENT SETUP menu use the up or down arrow keys to highlight EXIT then press the enter key To set additional parameters continue as detailed in the following text Time 1 Press the down arrow key to move the highlight to TIME and press the enter key EXIT EIT LOCKED OFF LOCKED orf LOCEIHG CODE dal LOCEIHG CODE dni LAHGUAGE English LANGUAGE English TIME 14 41 85 TIHE 14 4dEl 65 DATE 19 84 86 DATE 19 64 86 DATE FORMAT DOS MMS DATE FORMAT DOMM YY OPERATOR ID 506 OPERATOR ID Sha 2 Use the up and down arrow keys to set the highlighted digit of the time to the correct value Use the right and left arrow keys to highlight each digit in turn for adjustment with the up and down keys until the correct time is set 3 Press the enter key to confirm the settings and start the clock running from the set value Not
29. nm 668 UHITS EF Ll RESOLUTION B Bi 18 8 9 10 11 12 The number of decimal places RESOLUTION to which the measurements will be displayed can then be set at the next option in the MEASURE SETUP menu The fluorimeter will auto range resolution for measurements too large to be displayed in the set resolution EXIT METHOD ID ae HAME Chlororha11 EXCITATION nm 436 EMISSION nm 668 UHITE RFU RESOLUTION ERE To define the calibration parameters select the last option of the MEASURE SETUP menu CALIBRATION SETUP EIT EIT METHOD ID BS HO OF STDS 3 HAME Chlororhs11 STANDARDS TABLE ERCITATIOH nm 436 FLUORESCEMCE TABLE EMISSION nm 668 P M T GAINEHI B48 UHITE mail AUTO SET GAIH RESOLUT TOM 8 81 CAL REMINDER orf i CAL INTERWALCHRS dl Once in the CALIBRATION SETUP menu set the NUMBER OF STANDARDS that will be used at calibration The number can be set between 1 and 6 but this does not include the blank no fluorescence standard which is required for all calibrations The concentration values of the standards not including the blank can then be set in the STANDARDS TABLE EXIT STD ONE 14 888 STD TWO 20 858 STD THREE 36 886 The menu option below this is the FLUORESCENCE TABLE The default raw fluorescence values will be shown here until a calibration is successfully completed Once a calibration has been performed the raw fluorescence
30. of the highest grade of purity AR Grade and all should be checked for excessive background fluorescence at the analytical wavelengths 7 Samples and working standards should not be stored due to the effects of evaporation as well as chemical and photo degradation Only prepare samples and working standards when they are required for analysis 8 Fluorescence is inversely proportional to temperature Ensure that all samples and standards have equilibrated to ambient temperature before analysis If in doubt use a temperature controlled cuvette holder 9 Refrigerated or cold samples will form micro bubbles on the cuvette wall as they warm up These are a common cause of drift in readings Ensure all samples and standards have equilibrated to ambient temperature before analysis 2 10 Check the linear range for each method and where necessary use a multi point calibration or calibration curve 11 Be aware of the effects of quenching and where necessary use sample dilution or extraction methods to eliminate this 12 Monitor the blank during and between batches to identify any increase due to contamination 13 Sources of contamination to be considered include cleaning agents micro organisms particles in suspension stop cock grease filter paper residues and plasticisers leached from containers caps or sealing materials 14 Many fluorescent assays are pH dependent Ensure the pH of all samples and standards is within sp
31. of the MEASURE screen To confirm the save is successful the message Result Saved is shown briefly on screen Chlororhy1l1 0 28 Fesult Saved SYSTEM GAIN 66 EXIT SALE A maximum of 100 results can be saved in this way for each method If more than 100 results are required the results area must be cleared before the 101 result is taken Please see section Stored results Results mode for details of how to achieve this Auto save To remove the need to manually select SAVE after each result AUTO SAVE can be activated After every read operation either timed or manual the concentration will be automatically saved To ensure every result is saved select the required method as detailed in Using a stored method and then return to the MAIN menu Enter the SETUP menu and select the RESULTS STORAGE section Move to AUTO SAVE and set to On Using the EXIT options return to the MAIN menu and then move to the MEASURE screen Each read operation will now have the result automatically saved and this is indicated on screen by the status message Result saved Amaximum of 100 results can be saved in this way for each method If more than 100 results are required the results area must be cleared before the 101 result is taken Please see section Stored results Results mode for details of how to achieve this 26 Stored results Results mode 1 6 To access the saved results for t
32. onship is then linear and the equation can be written as F 2 3KQPo EbC The accuracy of fluorescent measurements is very high because the radiant energy being formed is measured directly There are also only a few easily controlled limits on the sensitivity of fluorescence measurements From the equation above it can be seen that adjustments made to instrument electrical noise and competing radiations along with physical limitations such as radiation energy sample volume and cell size affect the measurement sensitivity Section 4 Instrument Menu Options In this section a brief explanation is given for the main menu options available These will be covered in greater depth in the sections appropriate to their usage Measure Screen Chlororhyl1 003 Cal Fesuired SYSTEM GAIN 40 EXIT SALE METHOD NAME user set current method name or DEFAULT METHOD LAST READING result from last read or UNITS OF MEASURE as set in MEASURE SETUP menu SYSTEM GAIN 0 to100 as set up in MEASURE SETUP menu Note This option is not available on the Model 6270 CALIBRATION REMINDER flashing user alert indicating that a calibration is due TIMED READING COUNTDOWN number of seconds until the next auto read is performed READ allows the user to take a reading of sample standard currently in the sample chamber SAVE allows the user to save the last reading to instrument memory To exit this menu highlight EX
33. ose items defined in the maintenance procedures herein there are no user serviceable items in this instrument Removal of covers and attempted adjustment or service by unqualified personnel will invalidate any warranty and incur additional charges for repair Reference should always be made to the Health and Safety Data for any chemicals or reagents used All available information advice and warnings on the handling storage use and disposal of such must be carefully observed When not available this data must be requested from the supplier before proceeding in any way It is important that good laboratory practice is observed when handling samples chemicals reagents and ancillary equipment in order to carry out measurement and analysis with this instrument Suitable safety and personal protective equipment must be used at all times If it is suspected that safety protection has been impaired in any way the fluorimeter must be made inoperative and secured against any intended operation The fault condition must be reported to the appropriate servicing authority In all such reports the model number and serial number of the fluorimeter must be quoted Contents Section 1 Section 2 Section 3 Section 4 Section 5 Section 6 Section 7 Section 8 Section 9 Section 10 Section 11 Section 12 Section 13 Section 14 Section 15 Introduction Instrument description Good practice guidelines Getting Started Unpacking Ins
34. r Excitation filter Locking screw 6 2 28 Power on and Self Tests Connect the mains supply cable to the rear panel mains input socket and plug the other end into a suitable mains supply socket Lift the sample chamber lid on the instrument and ensure that there is no sample or other item present in the sample holder close the lid Switch on the supply socket then the instrument using the Power switch located on the rear panel The instrument will then perform a power on self test protocol The following screen will be shown for approx 3 seconds Please ensure sample chamber door remains closed during power on tests Followed by Barloworld 6285 Barloworld 6280 Fluorimeter Jenway Barloworld 6270 Fluorimeter Jenway Fluorimeter Jenway On successful completion of these tests the Main Menu screen will be displayed For optimum performance a 10 minute warm up period is required if the ambient temperature is below 10 C The unit must be re calibrated and sample measurement repeated if this calibration check shows excessive drift Should a problem occur during the self tests an information box or error support message will be displayed For assistance please refer to the troubleshooting section in this manual Example Menu Operation A common operating system is used throughout this and similar Jenway products a brief overview of navigating through the menu system with the cursor keys follows
35. r Supply 90 to 230Vac universal mains adapter 12Vdc input cigarette lighter cable and battery pack options Size 365 w x 272 d x 160 h mm Weight 6Kgs 33 Glossary of Terms Filters There are three types of filter commonly used in fluorimeters Band pass filters Cut Off filters and Interference filters The first two offer a low cost general purpose option while the more expensive interference filters offer precise selection of excitation or emission wavelengths Often one type will be used for excitation and another for emission depending on the proximity of interfering wavelengths An excitation filter is fitted between the light source and the sample to limit the wavelengths reaching it to those absorbed by the analyte but not by any other substance in the sample matrix An emission filter is fitted between the sample and detector to restrict the wavelengths reaching the detector to those due to the fluorescence of the analyte while blocking those due to scatter and stray light Band pass filters can be used to isolate excitation or emission wavelengths their broad transmittance spectra enable them to be used in many applications However when used for both excitation and emission great care must be taken that their transmittance ranges do not overlap Most useful of those available are the UG1 filter with transmittance from 320 to 380nm BG28 from 380 to 500nm and VG9 from 480 to 580nm Cut off filters low cut wil
36. rations can still be performed when the instrument is locked LOCKING CODE 3 digit code that must be entered to unlock the instrument LANGUAGE choice of English French German Italian and Spanish TIME set to time stamp readings and calibrations DATE set to date stamp readings and calibrations DATE FORMAT DD MM YY or MM DD YY lf the date format is changed to MM DD YY the changes are reflected straight away Used to identify saved printed results OPERATOR ID 3 digit code can be entered from 000 to 999 Filter Status Menu This menu provides a summary of the current method wavelength setting and excitation and emission filters From the MAIN MENU press the down arrow key to move the highlight on the display to cover FILTER STATUS then press the enter key The following display will be shown EXCITATION 425 METHOD SETTING 425 FILTER TYPE EP PART HO 627 124 EMISSION 618 METHOD SETTING 618 FILTER TYPE COL PART HO 627 127 EXIT EXCITATION vvavelength in nm of current excitation filter METHOD SETTING wavelength in nm set in MEASURE SETUP menu FILTER TYPE 3 types of filter are available BP Bandpass COL Cut Off Longpass and NPB Narrow Bandpass PART NO Jenway part number of filter module EMISSION wavelength in nm of current emission filter METHOD SETTING wavelength in nm set in MEASURE SETUP menu FILTER TYPE 3 types of filter are available BP Bandpass COL Cut Off Longpass and NPB Na
37. red and no mains power supply is used this indicates the battery level is low Further instructions on the Porti printer can be found in the operator manual included with the printer 5 Now to print the result displayed on the instrument screen just press the Print button on the fluorimeter 28 DataWay 1 2 10 Dae Connect the supplied cable between the PC and the instrument RS232 socket If using for the first time insert the DataWay CD into the CD drive of the PC The CD will automatically run and installation instructions will be shown on the PC screen Once installed the software can be opened either from the programs menu or with the DataWay icon on the PC desktop Open the DataWay software A pop up box will appear as the software starts up Tick the box to the left hand side of the instrument model number and then press the search button When the software has found the attached instrument the right hand side Connect button will be available Click on this to connect DataWay Once connected click the Ok and DataWay will fully open For further help or greater detail on using DataWay please consult the DataWay help file Click on the Data Manager tab of the DataWay screen Now to print the result displayed on the instrument screen just press the Print button on the fluorimeter or the take reading button in DataWay For instruction on saving data plotting graphs and other features of DataWay ple
38. rrow Bandpass PART NO Jenway part number of filter module EXIT allows the user to return to the previous menu 13 Section 5 Instrument Settings Note Using the left arrow key to move from the settings on the right hand side of the menu will abort any adjustments you have made and return the value or selection to its previous settings The enter key must be used to confirm any changes All settings listed below are accessed from the INSTRUMENT SETUP menu as shown below EXIT LOCKED OFF LOCEIHG CODE Gai LAHGUAGE English TINE 21 53 df DATE A382 ee DATE FORMAT ODS MMS YY OPERATOR ID BBH The 62 Series Fluorimeters offer a number of important functions to control or restrict use of the instrument This can be useful in developing GLP procedures or for controlling usage in multi user installations The instrument LOCKING CODE restricts access to the set up menus without the entry of a 3 digit security code When the LOCKED function is enabled settings can only be viewed not changed Calibrations can still be performed when the instrument is locked Instrument Lock 1 Use the down arrow key to select LOCKED from the menu and press the enter key 2 The instrument lock can be turned on or off using the up or down keys Press enter to confirm your selection 3 With the instrument lock set to Off all functions of the instrument are available to all users 4 With the instrument lock set to On all methods will be
39. state to a higher energy excited level This is known as excitation and the amount of energy transferred to the molecule or atom will depend on two main factors The composition of the matter under investigation and the energy and wavelength of the radiation have a significant effect on the transitions of electrons The molecule or atom converts the excitation energy to vibrational or light energy and the electron returns to its ground state Vibrational energy is transferred through movement and collision with other molecules but energy not lost in this way is released as light radiation The light emission is known as fluorescence and if some energy has been removed through vibration it will be of a lower energy and longer wavelength than the excitation energy The wavelength and intensity of the emitted radiation is dependent on the structure and composition of the molecule and the excitation wavelength used Relationship between concentration and fluorescence The fluorescence signal F and concentration C of the matter under investigation are related by F RQPo 1 10 YO Where K A constant characteristic of the instrument Including instrument electronics pH and Temperature Q Quantum efficiency Photons emitted Photons absorbed Po Power of incident radiation Molar absorptivity of the species matter b Absorption path length If the concentration of the matter in question is low dilute EbC is small The relati
40. t Display oe 1 Quinine 2 ee pe ie estors Sola P Cal Resuired 4 SYSTEM GAIH Sie ee hid READ SAWE 5 6 EXIT 1 Method Name this will appear on all measurement screens with the exception of Raw Fluorescence 2 Results display provides direct readout of standards and sample results 3 Units of measure shows selected measurement units ppm RFU U ml mU U I UM mMA M I ug ml mg ml ug di mg dl g dl ng l ug l mg l g l none ppb 4 Status message shows the current instrument status such as reading or printing and provides a reminder when calibration is due 5 Gain shows the current photomultiplier tube gain setting 0 100 Gain should be optimised either manually or by using auto set gain for each method Note This option is not available on the Model 6270 6 Menu Options EXIT allows the user to return to the main menu READ press read to read the sample or standard SAVE saves the currently displayed result to instrument memory 5 2 26 Rear Panel Layout DANGER UV radiation hazard Disconnect from power before opening L OCN Red sockets 3 1 Output Socket 9 way output socket for RS232 2 Switch Power On Off switch 3 Connection Sockets 4 x pin sockets for heated cell block controller 4 Power In Socket Connection socket for 12V DC universal mains adapter 2 27 Sample Chamber Filter and Cuvette Positioning Emission filte
41. tallation Fluorescence Theory Instrument Menu Options Instrument Settings Instrument Lock Date Time and Language Security Operator ID Setting up a method Using a stored method Calibration Performing Measurements Timed Readings Saving Results Printing Results Maintenance amp Troubleshooting General and routine maintenance Error codes and troubleshooting Accessories and Spares Optional accessories Specification amp Data Technical specification RS232 serial interface Glossary of Terms EC Declaration of Conformity Section 1 Introduction 1 1 Instrument Description Three models are available to cover a wide range of applications The model 6280 covers the most sensitive determinations with emission wavelengths up to 650nm Where higher emission wavelengths will be utilised the model 6285 with its red enhanced detector is applicable For less sensitive applications with a broader wavelength range the model 6270 will meet the necessary requirements All three models offer intuitive operation with a user interface based on logical menus that can be navigated from the simple keypad Up to 20 methods can be created without restriction and be saved for future use they can also be locked against accidental change by password entry ensuring data integrity The permanent time and date tag on every stored reading further enhances Good Laboratory Practice while calibration reminders and operator identity can also be
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