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BD 3096 Charcoal Grill User Manual

1. d Insert a screwdriver or a similar tool into the recess on the right side of the fuse holder push slightly and take the fuse holder out You will see two fuses L Take the faulty fuse s out Note You need both fuses for op eration Faulty fuses can typically be detected by a broken fuse wire in the glass envelope 91 Microplate Luminometer 92 Maintenance LI Insert new fuse s L Caution Observe the required fuse types current and fuse ratings see label on instrument rear panel Otherwise the operating safety cannot be guaranteed and the instrument will lose its UL Underwriters Laboratories approval LI Install fuse holder correctly so that the pin fits into the hole corresponding to the operating voltage Function check Lj Attach power cable to the wall outlet _J Turn instrument on L Check if green LED is on If the green LED is not on please contact our Service Depart ment Microplate Luminometer Figure 8 2 Power supply unit on the instrument rear panel CAUTION Maintenance 8 3 Changing the Operating Voltage for MONOLIGHT 3096 You need 1 small flathead screwdriver 1 flat nose pliers 2 fuses for the respective operating voltage For operating voltage 230 V Fuse 160 mA slow blow For operating voltage 115 120 V Fuse 315 mA slow blow Use only UL approved fuses The instrument is designed for operation at 230 V Euro voltage or 115 V US Voltage The voltage setting
2. System Description Detachable service door The service door is to the right of the instrument door You will need this door only if you are connecting the MONOLIGHT 3096 injector unit option see section 3 6 2 2 2 Microplate Loading Compartment The microplate loading compartment is a compartment that is separated from the measurement chamber it is accessible via the instrument door In its home position the mobile microplate transport unit is completely inside this compartment After the power is turned on and upon completion of a measurement the transport unit automatically moves to this position This movement can also be controlled via software The transport unit is a mobile tray accommodating the mi croplate Controlled via software the transport unit can be moved into the measurement chamber so that the wells are ex actly positioned below the photomultiplier one after the other The transport unit can also be moved by hand The transport unit comprises a spring loaded support tray for the microplate with a device that prevents its slipping out of place and a frame which can be folded up b Friction spring fixing the microplate a Fitting piece for microplate or plastic tub Drilled hole for transport securing screw c Spring loaded support tray for the microplate Microplate Luminometer CAUTION System Description The spring system ensures that different microplate types re gardless of their
3. General tab Select the type of microplate you are using 96 wells Select Take background at start of plate measurement if you want to run a background measurement prior to the sample measurement Raw Data PC Parameters Raw Data PC 57 Microplate Luminometer 58 Figure 5 5 EXCEL TM tab Raw Data EXCEL TM tab Create an EXCEL macro for control of a measurement proce dure by making the necessary entries here Prerequisite is that EXCEL 7 0 is installed on your PC and running Figure 5 5 Raw Data PC Parameters Raw Data PC ply Timing E General HE Excel TM E Comment Worksheet At cell p 1 Excel TM macro orea Auto send to Upon completion of the measurement the Excel macro is automatically sent to the Excel program and executed there Workbook Selection of workbook Click on lt gt to dis play the browser The subdirectory Sim plicity workbook is defaulted At cell The measured data are read starting at this cell of the Excel spreadsheets Excel TM macro Entry of an Excel macro Microplate Luminometer Figure 5 6 Input field for a comment on the measurement protocol Raw Data Comment tab The comment entered here appears after the protocol has been saved in the Protocol Manager next to the measurement proto col list when the respective measurement protocol is selected Raw Data PC Parameters Raw Data PC x E Timing E
4. Protocol file name Figure 4 3 Slow Kinetics PC Parameters Slow Kinetics PC Measurement protocol Slow Kin fics ECY Plate Timing E EX Well Timing E General EE EycelTh E Comment Parameters kag Total duration of experiment Plate Timing tab pim hms Hh Hm H No of repeat plate measurements J 4 Interval con ee h m Note Interval Total duration of esperiment No of repeat plate measurements Cancel As soon as you have entered your parameters click lt Save As gt to save the protocol under a new name or click lt Save gt to save it under the old name The stored measurement protocol is then filed in the field Runable protocols Anew measurement protocol does not include the lt Save gt but ton the file still bears the name of the protocol type which may not be used for saving this protocol 44 Microplate Luminometer Figure 4 4 Measurement menu Raw Data PC Presentation of microplate Structure of the Software 4 4 Measurement Menu Upon selection of the measurement protocols in the Protocol Manager and clicking on lt Run gt the program displays the measurement menu which is structured according to the se lected measurement method At this point the luminometer is initialized if this has not been done yet see section 3 9 4 4 1 Structure of the Measurement Menu Title bar with file name and name of measurement method Menu bar Tool bar Untitled Raw Data PC
5. L Type A spcsetup This will start the setup program and display the start screen L Click lt Next gt The Select Destination Directory dialog box appears and you can select the destination directory and the directory C Program Files Simplicity PC Figure 3 6 L Click lt Next gt if you want to keep the defaulted directory Simplicity 1 0 Alpha 6 Installation Select Destination Directory Please select the directo where Simplicity 1 0 Alpha tiles are to be installed Free Disk Space After Install is based on Your current selection of files to install A negative number indicates that there is not enough disk space to install the application to the specified drive C Programme Simplicity Browse Curent Free Disk Space 341652 k Free Disk Space After Install 336740 k Back Cancel Microplate Luminometer Figure 3 7 Ready to Install dialog box Installation and Commissioning Click lt Browse gt if you want to choose another directory The Select Destination Directory dialog box appears In the Path text box type the name of the new directory Confirm your entry with lt OK gt The new directory is created and the program returns to the Select Destination Directory dialog Dox Click lt Next gt The Ready to Install dialog box is displayed Figure 3 7 Click lt Next gt to install the program Upon successful completion of the installation process you w
6. Manager and you can select a new measurement protocol 39 Microplate Luminometer Structure of the Software 4 Structure of the Software 4 1 Software Structure The Simplicity software comprises three levels 1 The Protocol Manager which is displayed after program startup is the program s switchboard featuring the following functions e Create measurement protocols lt Create gt e Edit measurement protocols lt Edit gt e Delete measurement protocols lt Delete gt e Select measurement menu lt Run gt e Load data file lt Retrieve Data gt e Define instrument parameters Instrument tab e Define directory for measured data lt Options gt see section 4 2 2 Input of measurement parameters To enter measurement parameters either create a new measurement protocol in the protocol manager lt Create gt or edit an existing measure ment protocol lt Edit gt The respective protocol is loaded and can be filled out and saved for later measurements see section 4 3 3 The measurement menu is structured depending on the se lected protocol type Here the measurement is started and executed At the same time the measurement menu is the switchboard for display and further processing of the meas ured results result display graphical presentation and export of the data into EXCEL see section 4 4 section 4 6 includes an overview of the program functions 40 Microplate Luminometer P
7. Slow Kinetics Maximizing Minimizing graphs In the graph window grab the horizontal bar dividing the graph ics and data section with the mouse and pull it down Thus ad ditional items as well as the listing of the measured value with calculations become visible average peak etc You may also increase the graphics section by moving the horizontal bar up so far that the entire data section is hidden ME riiki Oma Geach Prod famia Tae a Pa Lise Past a bak Wet Bim es Spo m1 Hid H Ha Li E The graph window The curve trend of the measured values is presented in the graph window default setting automatic scaling The well po sitions and associated colors are listed in the graphics legend next to the diagram Options in the graph window Smoothing Smoothing the measured values Log scale Logarithmic scaling of the measured val ues RLU range Selection of scaling Automatic Default setting is automatic scaling of the measured RLU values comprising the top and bottom value of each coordinate Manual Manual setting option for the display area Upon selection of this item you may enter the start min and end max of the RLU ranges When you click Manual once more the entered range can be applied to the graphics 71 Microplate Luminometer Fast Kinetics 7 Fast Kinetics Creating a new measurement protocol Editing an existing measurement protocol Figure 7 1 T
8. ble Then the measurement is carried out for data point 2 etc The data can be presented as a graph or in a table or imported into an EXCEL spreadsheet for further processing 63 Microplate Luminometer Creating a new measurement protocol Editing an existing measurement protocol Figure 6 1 Plate Timing tab in 64 the Slow Kinetics dialog box Slow Kinetics 6 2 Measurement Protocol In the Protocol Manager select the protocol type Slow Kinetics and click lt Create gt In the Protocol Manager select an existing measurement pro tocol and click lt Edit gt The Slow Kinetics Parameters dialog box with several tabs is displayed and you can enter the measurement parameters Figure 6 1 Slow Kinetics PC Parameters Slow Kinetics PC ET Plate Timing ETA Mell Timing General EE Excell Tht E Corment Total duration of esperiment jonm him jh in Mo of repeat plate measurements p i Interval onm him Hote Interval Total duration of esperiment 7 Ho of repeat plate measurements Cancel Plate Timing tab The following mutually dependent parameters must be taken into account for the kinetics measurement e the number of data points e measurement time per well 1 sec e the number of samples to be measured e the number of injections per well e the delay time between injection s and measurement time e the expected duration of the light emission of the sam
9. so that the door is pressed into the rubber foam LI Align the service door exactly LI Press the service door down firmly on the right hand side LI Connect the MONOLIGHT 3096 and the injector unit using the supplied cable Insert the 15 pin connector into the MPL port in the injector unit Power switch Power cord Operating MPL port connector voltage indicator LI Check that the wall outlet voltage matches the operating voltage labeled on the injector unit If not change the oper ating voltage as described in section 8 14 LI Connect the injector unit to the wall outlet using the supplied power cable LJ CAUTION Always turn the luminometer on first then wait for at least 20 seconds before turning on the injector unit LI To stabilize reagent bottles you may use the supplied rea gent bottle holders made of stainless steel 25 Microplate Luminometer 26 Figure 3 6 Selection of destination directory Installation and Commissioning 3 7 Software Installation The Simplicity software includes an automatic installation program The Simplicity software is compatible only with PC format which must have Windows 95 or better installed See page iii for Typographical Conventions Installation of Simplicity PC 1 Close all Windows applications LI Insert program disk in drive A LI Select the Windows 95 command Run The Run dialog box appears
10. 800 825 5832 Luminometer packing material Cat No is 551484 and the Injector Unit packing material is Cat No 551485 Microplate Luminometer Maintenance 8 5 Servicing and Cleaning the Injector System This section describes service and maintenance work that can be carried out by the user on parts of the injector system in cluding the MONOLIGHT 3096 injector unit the injector tubings to and from the MONOLIGHT 3096 and the injector tips in the MONOLIGHT 3096 8 5 1 Regular Maintenance and Checks e After use rinse the pump thoroughly with distilled or deion ized water Section 8 7 e lf the pump is not used for a short time it should be primed with distilled or deionized water e Do not operate the pump without liquid e Do not bend the injector tubings e Check screw fittings of injector tubings regularly for leaks at the injector head and at the pump head If any leaks occur determine the cause loose or defective screw fitting Tighten or replace fitting see section 8 10 e Check injector pumps for leaks If liquid escapes below the pump head tighten syringe If liquid escapes at the syringe replace the syringe section 8 12 If there is a leak in the pump head you must replace the pump contact Service 8 5 2 Cleaning the Injector Unit outside The surface of the injector unit is protected by a sturdy wash able varnish If it is dirty or dusty clean it using a moist cloth If necessary use a m
11. Any printer supported by Windows Software requirements e MS Windows 95 e MS EXCEL 7 0 optional I7 Microplate Luminometer Figure 2 10 Structure of Simplicity PC software 18 System Description 2 6 Software The MONOLIGHT 3096 Microplate Luminometer is operated by the software package Simplicity Photon Counter Simplicity PC designed specifically for this instrument The program comprises three levels Protocol Manager management of protocols and system pa rameters Measurement parameter input depending on the selected measurement protocol Measurement menu the measurement is performed as de fined by the selected protocol type and the entered parameters and the results are displayed Protocol Manager Total duration of experiment m on 4 fe No of repeat plate measurements Note Interval Total duration of experiment No of repeat plate measurements Save Save As Cancel sae Parameter input Measurement menu Microplate Luminometer Installation and Commissioning 3 Installation and Commissioning 3 1 Overview Proceed as follows to prepare your first measurement Step Section Page Unpack and set up the instrument 3 4 20 Remove transport safety screw 3 3 z Check microplate transport unit manually Connect injector unit Connect injector tubings to pumps Connect reagent bottles to pumps i a 24 Connect MONOLIGHT 3096 and injector unit
12. Confirm the file name with lt OK gt The program returns to the Protocol Manager and shows the name of the new measurement protocols in the field Runable protocols Li In the field Runable protocols in the Protocol Manager select the new measurement protocol and click lt Run gt Then the measurement menu is displayed Figure 3 15 Microplate Luminometer Installation and Commissioning Figure 3 15 Untitled Raw Data PC _ oy x Raw Data PC File Edit Options Measure View Help Measurement menu 0A S Se Sle Leip lee al Background 0 Date Subtract Time CUA APL Measurement duration 1 02 ee B BE BE 1 l M Measuring the entire microplate O Click the button Measuring a selected area LI Click on the first well of the area you want to measure and with the mouse button held down drag the cursor over the wells to be measured These wells are highlighted by a dark color Figure 3 16 Release the mouse button when the desired area is marked Figure 3 16 Selected well area J Click the Ics button Then the selected wells turn white the others are gray shaded Figure 3 17 ir Microplate Luminometer Installation and Commissioning SSS E S Figure 3 17 mus a 2 z aj s ej 7 aj a w Defined well area A a x L The marked area is cleared again when you define a new area with the mouse and click the ics button again Running a measurement LI Click to start the
13. Dual Measurement If this item is checked each selected sample is measured twice and both measured values are displayed in the respective field This function can be used with or without injectors Parameters must be entered accordingly No injector Delay time 1 measurement gt 2 measurement Injector 4 Delay times Inj 1 gt 1 measurement gt 2 measurement Delay times 1 measurement gt Inj 2 gt 2 measurement Injector 1 amp 2 Delay times Inj 1 gt 1 measurement gt Inj 2 gt 2 measurement 68 Slow Kinetics Figure 6 5 General tab in the Raw Data PC Parameters dialog box Figure 6 6 EXCEL TM tab General tab Select the type of microplate you are using 96 wells Select Take background at start of plate measurement if you want to run a background measurement prior to the sample measurement Raw Data PC Parameters Raw Data PC EXCEL TM tab Create an EXCEL macro to control a measurement by making the necessary entries here Prerequisite is that EXCEL 7 0 is in stalled on your PC and running Figure 6 6 Slow Kinetics PC Parameters Slow Kinetics PC 69 Microplate Luminometer 70 Figure 6 7 Input field for a comment on the measurement protocol lt Save As gt Slow Kinetics Auto send to Upon completion of the measurement the Excel macro is automatically sent to the Ex cel program and executed there Workbook Sel
14. File Edit Options Measure View Help al Ee fez 3 2 Background 10 Date 01729799 Injection 1 volume 10 Injection 2 volume 10pl Subtract Time 08 47 17 aea p ie x A A E ee ee E E as sample matrix Status bar Tou must initialize the injectors before vou can measure Title bar shows the measurement method Menu bar shows the titles of the pulldown menus Tool bar shows the tool buttons you can work with in the displayed measurement mode 45 Microplate Luminometer 46 Structure of the Software Measurement Parameters The date time background injectors used and the measurement time are displayed below the toolbar You can choose if the displayed background value is to be sub tracted from the measured values Sample matrix The microplate is depicted as a sample matrix with each well being represented by a box Here you can select the wells to be measured For Raw Data the measured results are entered in the respective boxes for Slow Kinetics and Fast Kinetics meas urements are displayed in a separate ta ble Status bar The status bar keeps you informed about the status of the sample drawer and the next operating steps in accordance with the selected measurement protocol 4 4 2 Working with the Measurement Menu This section briefly describes all functions that all measurement methods have in common The functions which all measure ments with injectors have in common
15. General l EE Excel TM l Comment l Cancel The following buttons appear on all tabs of the entire measure ment protocol lt Save As gt lt Save gt is selected to define a new file name This is necessary in order to create a new measure ment protocol or to save an existing measure ment protocol under a new name Clicking on this button opens the Save As dialog box Enter a file name which differs by at least one character from the file name of the selected protocol type As soon as the protocol has been saved the program returns to the Protocol Man ager and shows the name of the new meas urement protocols in the field Runable proto cols saves the parameters in the existing measure ment protocol file After you have saved the pro tocol the program returns to the Protocol Manager 59 Microplate Luminometer 60 Figure 5 7 Measurement menu Raw Data PC Status bar Raw Data lt Cancel gt The changes are disregarded and the old values are still valid The program returns to the Proto col Manager 5 3 Measurement As soon as you have selected measurement protocol type Raw Data PC and you have clicked lt Run gt the program changes to the Raw Data PC measurement menu Communication with the Microplate Luminometer is established if this has not been done during program start Initializing instrument appears on the status bar and the yellow LED lights up Unti
16. by clicking tel The graph display is on top of the table window Figure 7 10 Measurement menu Fast Kinetics with graphical and tabulated result display e To display the curve trend of one or several selected sam ples select the wells in the sample matrix with the cursor and click E e To view the graph and table window on the screen at the same time click or I depending on the layout you prefer Figure 7 10 e To open or close one display mode click the button E or EE The remaining window will fill half the screen 87 Microplate Luminometer Figure 7 11 Graph window Slow Kinetics partial view Here you may pull the 88 dividing bar up or down with the mouse Fast Kinetics Further options for opening closing arranging maximizing mini mizing the size of graph and table windows Buttons in the upper right corner of the dialog box I Maximizing the windows to full size Ei Minimizing the windows to half the screen size Buttons in the upper left corner of the dialog box a Opens a pulldown menu including the item Close to close the table window Opens a pulldown menu including the item Close to close the graph window Maximizing Minimizing graphics In the graph window you can grab the horizontal bar dividing the graphics and data section with the mouse and pull it down Thus additional items as well as the listing of the measured value with the calcu
17. finished when all predefined measure ments have been carried out Figure 6 10 an mata tTepegrpepepc_efeyfea Fg LES E P E BLA Eo ENE Fua m Miis ma lid Li jie 73 Microplate Luminometer 74 Slow Kinetics Procedure with injections LI Select the wells to be measured LI Place plastic tub in plate loading compartment O Initialize injectors 24 LI Prime tubings if necessary at Insert microplate LI Insert microplate correctly in luminometer and close door BA O If necessary run background measurement 2 LI Select the wells to be measured E O Start measurement LI Data point 1 The pre defined injections and the measure ments of the selected samples are carried out in succession and the measured values are displayed on the screen If you have selected too short intervals the system corrects the in tervals using the actually required time LI Then the measurements at data point 2 are performed and the results displayed All following measurements are per formed in the same manner LI The measurement is finished when all predefined measure ments have been carried out Figure 6 10 Microplate Luminometer Figure 6 11 Measurement menu Slow Kinetics with graphical result display Slow Kinetics 6 4 Result Display Graphical and tabulated display of the measured results As needed you can call the window for graphical display of the measured data before during
18. for very fast reac tions requiring at least one injection during measurement The MONOLIGHT 3096 was designed to be used with the add on injector unit for measurement requiring injection in the instru ment This unit allows injection of up to two substances in the instrument even during measurement The MONOLIGHT 3096 Microplate Luminometer is equipped with a highly sensitive light detector A special measuring ge ometry ensures high sensitivity and a dynamic range covering more than 6 decades A special light guide system reduces crosstalk light transfer between adjacent samples to a minimum Crosstalk can be reduced further by selecting suitable microplates The sophisticated mechanical construction of the instrument offers further advantages for the user The microplate is placed on a mobile transport unit outside the measurement chamber and moved into the measurement chamber for measurement by means of a highly precise control mechanism that positions the sample wells exactly below the photomultiplier The separation of microplate loading compart ment and measurement chamber ensures precise measure ments simple operation and little maintenance and cleaning Microplate Luminometer Optional injector unit Windows based operation Three measurement modes System Description The MONOLIGHT 3096 is the basic unit which can be upgraded with the injector unit containing 2 injectors The MONOLIGHT 3096 is designed for work
19. height will always be pushed directly against the frame and thus as close as possible against the photomul tiplier Since the sensitivity decreases with the squared distance to the detector the spring system significantly increases the sensitivity of the measuring system If the MONOLIGHT 3096 is operated with injectors a small plas tic tray is supplied with the instrument Put this plastic tray into the microplate loading compartment instead of the microplate when priming and washing the tubing to collect the liquid es caping from the injector tips Keep microplate loading compartment clean e Keep frame and loading tray clean and free of dust If neces sary clean it with a moist cloth Dirt will also have an adverse effect on the measured result e lf liquid does get under the microplate transport unit you can move this unit by hand or via software into the measurement chamber so that the bottom of the loading compartment can be cleaned e Always make sure that liquid does not get into the measure ment chamber How to insert a microplate _J Make sure that the service door next to the instrument door is closed properly L Open the instrument door of the Microplate Luminometer LJ Turn up the frame of the transport unit You cannot open the frame if the transport unit has been moved a bit to the right towards the measurement chamber In this case move it by hand all the way into the loading compartment 1 Place the
20. measurement The defined measure ment procedure begins the individual wells are selected in succession and measured using the predefined measure ment time The results are displayed in the well matrix on the screen These are net results provided a background meas urement has been carried out prior to the measurement display of the BG values in the field Background and the item Subtract has been activated L Click I to enter a comment for the measurement which will be printed out in the report This opens the Measurement Comment dialog box and you can enter your text 3 Click l to save the measurement data and then enter a file name 38 Microplate Luminometer Installation and Commissioning Next measurement i To run the next measurement with the same measurement protocol click New measurement The next measure ment can be started immediately when the data of the preceding measurement have already been saved Otherwise you will be prompted to save this data If you have stored the data or if you have replied lt No gt the screen is cleared for the next measurement and the icon buttons are enabled again Li To run the next measurement with another measurement protocol select Exit on the File menu If the data of the preceding measurement have not yet been stored you will be prompted to save this data After you have stored the data or if you have replied lt No gt the program returns to the Protocol
21. must be changed only when operating the instrument in a country with a different operating voltage Power cord connector Power switch Recess for screwdriver Fuse holder L Turn main switch to off 0 _ Unplug power cord from wall outlet _J Insert a screwdriver or a similar tool into the recess on the right side of the fuse holder push slightly and take the fuse holder out L Replace both fuses and carefully put the fuse holder aside 93 Microplate Luminometer Maintenance Figure 8 3 Power supply unit with fuse holder taken off Voltage selection card with pin L Remove the fuse holder to gain access to the voltage selec tion card Pull out the voltage selection card horizontally using a small pair of pliers to hold it at the white pin L Four operating voltages are printed on the voltage selection card and labeled accordingly Take the pin out and position it in the recess so that it is on one line with the correct operat ing voltage see Figure 8 4 Figure 8 4 Voltage selection card Recess Instrument lt lt Porz Operating voltage Indication pin To change the operating voltage to 120V you must position the indication pin as illustrated in this drawing and insert the card in this position again pushing it in the direction of the arrow printed side facing the power cord connector 94 Microplate Luminometer CAUTION Maintenance Ll
22. prepared microplate onto the loading tray such that the A1 well is in the upper right hand corner Microplate Luminometer 10 Measuring range System Description _J Turn down the frame and push it shut until it snaps into place audible click The microplate is held in place by the fitting piece Figure 2 4 a by the spring on the frame at the rear between the hinges b and by the spring loaded support tray c LI Close the instrument door Push it shut until it clicks into place with an audible noise Then the instrument is closed light tight 2 2 3 Measurement Chamber The measurement chamber is separated from the microplate loading compartment It is sealed light tight only when the serv ice door has been closed properly For measurement the transport unit with the microplate will move into the measurement chamber so that the well to be measured is exactly positioned below the photomultiplier The wells are measured column by column A1 to H1 A2 to H2 A12 to H12 For information on the position of the injector tips please read section 2 2 6 The measurement chamber is not accessible to the user Dirt in the measurement chamber can therefore be removed only by authorized service engineers 2 2 4 Light Detector The light detector measures visible light in the range from 300 to 650 nm The photons emitted by the sample are converted into electrons and multiplied by a photomultiplier The single pulses
23. range More than 6 decades Hardware Microcontroller for control of all instrument functions Injector Ports 2 at measurement position and 2 in the neighboring well positon Measuring Time About 100 s per microplate depending on the measurement protocol Scan Pattern Random selectable single well access by mouseclick Comm Ports Serial interface RS232 port for PC CAN bus connection to injector unit Dimensions W 400 mm D 545 mm H 200 mm Weight Approx 17 kg Operating Voltage 230 V 50 Hz 115 V 60 Hz Power Consumption 35 VA Temp Range Storage 0 40 C Operation 10 35 C Humidity 10 80 no condensation Regulatory mM pd Microplate Luminometer Technical Data Injection Unit Tubing Chemically inert PTFE tubing and connections PTFE or KEL F inter changeable liquid handling system and tips W 158 mm D 330 mm H 190 mm Power Consumption 25 VA PC Software Optional Software Microsoft Excel 97 Standard Configuration Protocol Manager Raw Data Available Additions Fast Kinetics Slow Kinetics Windows Features Fully resizable grids non contiguous areas selectable multiple document interface resizable zoomable graphs status bar draggable tool bars etc 111 112
24. see section 3 9 Instrument Configuration In this case communication with the luminometer is checked when the program is loaded When the communication has been established the program is fully operational Otherwise you cannot call the measurement menu Microplate Luminometer Figure 3 9 Instrument tab Figure 3 10 Instrument Options dialog box Installation and Commissioning 3 9 Instrument Configuration For instrument configuration select the Instrument tab in Protocol Manager and make the following settings see Figure 3 9 Simplicity Protocol Manager Photon Counter E Protocols Instrument Instrument options Check instrument Read configuration Background E Take background Options Help About E sit LI Click lt Instrument options gt The Instrument Options dialog box appears Figure 3 10 Instrument Options T Log gerial communication Serial port EOM Check instrument at startup Take background at startup Cancel L Select the communication port used in the field Serial Port COM1 or COM2 In most cases COM 1 is the proper choice rarely COM 2 If you select the wrong communication port the error message The instrument does not respond is displayed In this case you must select the other port 29 Microplate Luminometer 30 Installation and Commissioning L Check instrument at startup Select this item if you wan
25. the individual injections here Select injector inj 1 or inj 2 by clicking on the respective box You can then enter the injector parameters Please note Injector 1 injects into the well before the meas urement position injector 2 in the measurement position Volume Enter the injection volume in ul via keyboard or by clicking on the arrow buttons Injection 1 to measurement delay Enter the delay time be tween the injection of injector 1 and the start of the measure ment in seconds The hardware dependent minimum value of 2 05 seconds is defaulted and can be increased Injection 2 to measurement delay Enter the delay time be tween the injection of injector 2 and the start of the measure ment in seconds The minimum value of 0 seconds is defaulted and can be increased Raw Data PC Parameters Raw Data PC a X E Timing General l EE Excell Thi E Comment Note All delays are between the start of the respective events W Use injection 1 Volume hoo w y Use injection 2 T Use dual measurement Injection 1 to measurement delay 1 23 Sis Measurement duration 1 0 E e e 55 Microplate Luminometer Raw Data When using two injectors Select both injectors The following dialog box appears F ig ure 5 3 Raw Data PC Parameters Raw Data PC E x Timing tab ED Timing General ERE Excell Th E Comment Selection of Note All delays are between the start of
26. the respective events injector 1 and 2 W Use injection 1 Volume f D pl ul M Use injection 2 Volume fo pl su T Use dual measurement Injection 1 to injection 2 delay f 2 5 g s Injection 2 to measurement delay oo 3 Measurement duration f 0 amp amp 3 Volume Enter the injection volumes in ul via keyboard or by clicking on the arrow buttons Injector 1 Injector 2 Injection 1 to injection 2 delay Delay time between the injec ez P tions of injector 1 and 2 The hardware dependent minimum LLLI value of 2 05 seconds is defaulted and can be increased tp Injection 2 to measurement delay Enter the delay time be tween the injection of injector 2 and the start of the measure ment in seconds 0 second is defaulted and can be increas ed tv Example Programming a measurement with 2 injections bt r Pall P _ pre injection injection M measurement tp delay between pre injection and Qn injection tu delay between i injection and start of measurement Ty duration of measurement 56 e Luminometer Dual Measurement Figure 5 4 General tab in the Raw Data PC Parameters dialog box The Simplicity 1 0 software includes four pre programmed dual luciferase reagent DLR protocols Dual measurement proto cols cannot be created using the Simplicity 1 0 software The Use dual measurement option is grayed out in the screen seen in Figure 5 3
27. the syringes move the syringe pistons to the extended position and hold them there To do this you must use a cleaning protocol Therefore create your own cleaning protocol from the protocol you are using for measurement set the injector volumes as 150 ul and then save the protocol under another name e g cleaning protocol_acb 99 Microplate Luminometer 100 Maintenance Proceed as follows 1 1 1 1 2 1 3 1 4 ES 1 6 1 7 2 2 1 2 2 2 3 2 4 2 5 2 6 2 1 2 8 3 2 3 3 Wash with deionized or distilled water Use deionized or distilled water to remove the reagents remaining in the injection system Place the priming container on the microplate loading area Start the cleaning protocol and initialize the injectors Click the lt Priming gt button and select the injector s you want to clean select a volume of 150 ul and 20 injection cycles strokes Click lt OK gt to start the wash process Repeat steps 1 4 1 6 1 Cleaning Step Cleaning the Injector s with 0 1N NaOH Remove front cover of injector unit to watch the syringe movement Fill the reagent bottles and injector tubing with 0 1N NaOH Click the lt Priming gt button and select the injector s you want to clean select a volume of 150 ul and 20 injection cycles strokes Click lt OK gt to start the cleaning process Repeat steps 2 3 2 5 Start a measurement by clicking the green lt Start gt button click th
28. this end additional injector tubings fitting spare parts and a special tool for expanding the tubings are supplied as extras a Connection at pump head Figure 8 7 Injector unit Injector 1 Injector 2 Locking door Pump OUT IN Q Turn injector unit off LJ Pull off the locking door of the injector unit vertically from above Q Push tubing fitting screw over the tubing such that the threading is facing the tubing end Then push a washer over the tubing Figure 8 8 LI Using the special tool supplied expand the tubing end by pushing the tubing over the tool s arbor Figure 8 8 Pump T piece Washer tubing fitting screw Manufacturing the Tubing tubing connection m LJ Push T piece into expanded tubing end LI Insert tubing end with T piece into threaded borehole of pump head turn fitting screw with washer into the threading of the pump head and finger tighten it 102 Microplate Luminometer Maintenance b Connection at injector head Figure 8 9 MONOLIGHT 3096 Measurement head Injector head open Q Turn MONOLIGHT 3096 off and disconnect it from the wall socket LJ Open instrument door of MONOLIGHT 3096 open screws of service door and take door off Figure 8 9 Q Push tubing fitting screw over the tubing such that the threading is facing the tubing end Figure
29. to the MONOLIGHT 3096 and up to the reagent bottles which are placed on the tray onto the front side of the injector unit Pass the tubings through the respective openings and make sure you dont damage or squeeze them see Figure 2 8 15 Microplate Luminometer System Description The front side of the injector unit includes a detachable tray for the reagent bottles On the rear panel you have access to the following ports Figure 2 9 Rear panel of power switch power cord connector injector unit with connection ports EEE EEEE sueune e A Voltage selection switch Connection to 115 V 230 V MONOLIGHT 3096 2 4 Microplates All 96 well microplates in standard format can be measured on the MONOLIGHT 3096 Microplate Luminometer To obtain reliable accurate results you should only use non transparent opaque microplates Black microplates show the lowest crosstalk but they absorb light i e they are not suitable for samples with low light intensity White microplates are well suited since they reflect the light from the samples and thus increase the sensitivity but may increase background For general use we recommend the Falcon black opaque microplate Cat No 01 05500 16 Microplate Luminometer System Description 2 5 PC Minimum requirements e 486 PC or laptop e minimum 16 MB RAM e 200 MB hard disk e 3 5 disk drive e Color graphics card e Color monitor e Keyboard and mouse
30. ul Injection 1 to injection 2 delay f wad 3 m amp lt 3 Injection 2 to measurement delay o o 3 m 3 E Measurement duration E z Hes rea Volume Enter the injection volumes in ul via keyboard or by clicking on the arrow buttons Injector 1 Injector 2 Injection 1 to injection 2 delay Delay time between the injec tions of injector 1 and 2 The hardware dependent minimum eg value of 2 05 seconds is defaulted and can be increased tp Lit Injection 2 to measurement delay Enter the delay time be tween the injection of injector 2 and the start of the measure Microplate ment in seconds 0 second is defaulted and can be increased tv 1 Example Programming a measurement with 2 injections A r Pall P _ pre injection injection M measurement tp delay between pre injection and 2 injection tu delay between pe injection and start of measurement Ty duration of measurement 80 Fast Kinetics Figure 7 4 General tab in the Fast Kinetics PC Parameters dialog box Figure 7 5 EXCEL TM tab General tab Select the type of microplate you are using 96 wells Select Take background at start of plate measurement to run a background measurement prior to the sample measurement Fast Kinetics PC Parameters Fast Kinetics PC EXCEL TM tab Create an EXCEL macro to control a measurement by making the necessary entries here Prerequisite is t
31. until the next background measurement is performed It is also displayed in the measurement menu Select Subtract to subtract this value from the measured sample values te Luminometer Installation and C Micropla ioning L Click on lt Instrument Info gt on the Instrument tab to view the instrument configuration Figure 3 11 The current instrument parameters are displayed such as number and size of the installed injector The parameters cannot be changed Please contact BD Biosciences Pharmingen if you want to change the RLU factor Figure 3 11 Instrument Info dialog box L To define the subdirectory to which the measured data are to be saved automatically click lt Options gt on the Instruments tab and then enter the respective path name 31 Microplate Luminometer Figure 3 12 Registration Form a2 dialog box Installation and Commissioning 3 10 Software Registration The Simplicity PC software can be used for 30 days without registration after initial installation During this time you will be prompted to enter a registration password any time you call a protocol or go to the measurement menu see Figure 3 12 During this time you can close the Registration Form dialog box by clicking lt Run Now Register Later gt and then continue working with the protocol Registration Form Fast Kinetics PC Protocol for gt Microplate Luminometer Koo BO PharMingen All Rights reserve
32. which are digitally counted are directly proportional to the emit ted light quantity The light emitted by the samples can only be read from above and is since the photomultiplier for practical reasons flat in strument design is installed horizontally reflected by a mirror by 90 nearly without any loss Microplate Luminometer System Description RLU In contrast to other physical units luminescence is not indicated in fixed units of measure for example Lumen but in Relative Light Units RLU Therefore only results from the same lumi nometer type can be compared with each other An RLU factor has been defined in order to compare results from different lu minometers This factor multiplies all results Typically the RLU factor is 1 Upon request this RLU factor can be changed 2 2 5 Background Measurement and Subtraction The background is comprised of two components instrument background and reagent background The reagent background is obtained by measuring reagents without any analyte For example with a reporter gene assays this measurement is of the reagents measured without cell lysate 2 2 6 Injector Compartment behind the Service Door When your system includes injectors the injector compartment of the MONOLIGHT 3096 located behind the service door next to the instrument door is equipped as follows The service door is located to the right next to the instrument door On the right hand side it is he
33. 10 104 Microplate Luminometer Maintenance LI Insert injector head the sloping part is facing you Fit the in jector tips into the boreholes in the MONOLIGHT 3096 and the adjusting bolts in the recesses in the injector head Figure 8 13 LI Insert fixing screws and tighten them LI Close service door properly light tight see also page 20 Figure 8 13 Installing the Boreholes for fixing screws Measurement head injector head O 0 Injector head front view Boreholes for injector tips Adjusting bolts 105 Microplate Luminometer Maintenance 8 12 Replacing Pump Syringes When the pump syringe is defective e g leaky you must re place it Figure 8 14 Design of Cavro pump Pump head Syringe with knurled screw Piston Syringe piston holder LJ Empty pump by running a wash cycle with several strokes without having a bottle with liquid connected see section 0 Q Turn injector unit off and disconnect it from wall socket LJ Pull of locking door of injector unit vertically from above LJ Open knurled screw at syringe until you can detach it from the pump head and push it down with the syringe piston holder LJ Now you may take off the syringe from the front LI Insert new syringe in the syringe piston holder push it up and fix it with the knurled screw to the pump head LI Close injector unit 106 Microplate Luminometer Figure 8 5 Power supply unit o
34. 3 6 25 using a data cable Check operating and main voltage Connect injector unit to main and turn it on 3 6 25 Install Simplicity PC Start Simplicity PC Define interface in Simplicity PC Register software 310 32 32 First measurement Enter parameters Start measurement 3 11 36 19 Microplate Luminometer 20 CAUTION Cardboard box Check shipment Installation and Commissioning 3 2 Setup Site The MONOLIGHT 3096 Microplate Luminometer must be set up in a dry fairly dust free room and protected from direct sunlight and significant temperature fluctuations Do not set it up next to a radiator 3 3 Space Required The luminometer is rather small MONOLIGHT 3096 W xD xH 39 cm x 49 cm x 13 5 cm The instrument rear panel must always be at least 10 cm away from the wall or other devices to ensure that the cooling element can work properly and you have easy access to the power switch When using the injector unit option you must allow for an additional 20 cm width 3 4 Unpacking The luminometer is shipped in a cardboard box that contains two foamed inserts to protect the instrument against damage It also includes a smaller box with cables power cable connec tion cable PC Luminometer and the software The injector unit option is also delivered in a separate box The cardboard box is reusable It is recommended to store the box and packaging material for future shipping However boxes ca
35. 8 10 Then push a washer over the tubing LI Using the special tool supplied expand the tubing end by pushing the tubing over the tool s arbor Figure 8 10 Pump T piece Washer Banjo bolt Tubing Manufacturing the tubing connection LJ Push T piece into expanded tubing end LI Insert tubing end with T piece into threaded borehole of pump head turn fitting screw with washer into the threading of the pump head and finger tighten it 103 Microplate Luminometer Maintenance 8 11 Replacing Injector Tips The injector tips are located in the injector head behind the service door of the MONOLIGHT 3096 Figure 8 11 MONOLIGHT 3096 Measurement head Injector head open LJ Turn MONOLIGHT 3096 off and disconnect it from wall socket LJ Open instrument door of MONOLIGHT 3096 open screws of service door and take door off LI Open the 2 fixing screws of the injector head and take off the injector head directly behind the measurement head Figure 8 12 Injector head Injector tubing Injector head front view Position of injector tip in injector head Injector tip LI Unscrew the respective tubing fitting screw made of metal with tubing and pull it out LJ Push out injector tips towards the top LI Insert new injector tip and screw fitting screw into the injector head see also the previous section 8
36. 825 5832 Ll If you have not done so yet connect the computer to the printer and connect both devices to the wall socket 22 Microplate Luminometer CAUTION Figure 3 3 Microplate loading compartment and injector both devices open Installation and Commissioning 3 6 Connecting the Injector Unit The injector unit is supplied separately Connect the injector unit to the MONOLIGHT 3096 and then plug it in Afew simple steps suffice to connect the injector unit since the injector tubings are already installed to the injector head of the luminometer and the other end of the tubes includes the pre fabricated pump con nection U First unplug the MONOLIGHT 3096 Luminometer from wall socket LJ Put the injector unit to the right of the MONOLIGHT 3096 and pull the locking door of the injector unit off from above to ex pose the injector pumps LI Open the instrument door of the MONOLIGHT 3096 LI Hold the service door on the left side and pull it off the Velcro fastening You will find 2 rolled up injector tubings Microplate loading area Injector head Injector unit P S Tray for reagent bottles Microplate Luminometer Injector Unit 23 Microplate Luminometer Installation and Commissioning Q Pass injector tubing 1 to injector pump 1 and finger tighten the fitting screw in th
37. If not you must change the operating voltage by re plugging the voltage selection card so that the operating voltage matches local voltage See section 8 3 Check and replace the fuses if necessary See section 8 2 21 Microplate Luminometer Installation and Commissioning Figure 3 2 Instrument rear Fuse holder with panel indication of a r operating voltage w k PC connection Connection injector unit Power switch Power cord Fuse holder White pin at 115 V L Connect the luminometer and your PC or laptop to the serial communication port using the supplied connection cable which is provided with female connectors on both ends L1 When the wall socket and operating voltage match connect the luminometer to the wall socket using the supplied cable L Turn the Microplate Luminometer on at the power switch in strument rear panel The green LED 1 from left lights up and signals that the instrument is ready for operation L Perform a quick hardware test to check the microplate trans port unit open the instrument door and move the microplate transport unit by hand away from the home position Now turn the instrument off and on again After power is on the mi croplate transport unit moves to the home position audible and with open the instrument door visible movement If the transport unit does not move to this position please contact the Technical Service Department at 1 800
38. Injection 1 volume 10 Background o Date 01729739 Bo Injection 2 volume 10l Time jos 25 37 Measurement duration 1 0 Integral y You must initialize the injectors before you can measure Selecting the wells to be measured 0 Mark the well group you want with the mouse and press the Ics button to confirm your selection Li The selected wells are presented white and the measure ment value table is displayed according to the selected wells with data points see Figure 7 8 Microplate Luminometer Fast Kinetics Figure 7 8 Fast Kinetics PC Untitled Data Table E ea menu File Edt Options View Measure Window Help Fast Kinetics with HE FE be S EI S12 ee e E selected well groups Background O Date 01 28 99 Measurement duration 1 0s T Subtract Time 09 26 52 a Integral Result table with data points corre sponding to the pre defined meas urement times nmn TAN Fress the go button or space bar or enter to measuring Please note Before running a measurement if desired select the autosave mode Select this function on the Options menu Procedure without injections LI Insert microplate correctly into the luminometer and close cover LJ Run background measurement if necessary Beh LI Select the wells to be measured h Q Start measurement h Q Each sample is measured according to the pre defined measurement time and the measured values are displayed per data p
39. Insert the voltage selection card as shown in Figure 8 4 pushing it into the instrument in the direction of the arrows with the printed side facing the power cord connector Li Adjust the indication pin in horizontal position LI Insert fuse holder again properly so that the indication pin fits into the hole of the fuse holder with the proper operating volt age It must work smoothly otherwise the operating voltage is not set correctly Function check J Attach power cable _J Turn instrument on L Check if green LED is on If the green LED is not on please contact our Service Depart ment Use only UL approved fuses Otherwise the operating safety cannot be assured and the instrument will lose its UL ap proval 95 Microplate Luminometer 96 Maintenance 8 4 Taking the Instrument out of Service for Transport L Move microplate transport unit to home position for example by turning the Microplate Luminometer off and on L Open instrument door L Screw in red transport securing screw pass it through the in strument frame into the drilled hole in the microplate transport unit to arrest the transport unit _1 Close instrument door L Turn luminometer off and disconnect it from main L Put luminometer in the original shipping cardboard box with the two foamed inserts Additional cardboard boxes including foam inserts are available for purchase Contact BD Bio sciences Pharmingen Technical Services at 1
40. MONOLIGHT 3096 Microplate Luminometer Operating Manual BD Biosciences www bdbiosciences com F Clontech Immunocytometry Systems Transduction Laboratories a Discovery Labware PharMingen terj For research use only Not for use in diagnostic or therapeutic procedures Not for Resale l BD Biosciences Asia Pacific Brazil Canada Europe Europe Japan Mexico United States Tel 65 8610 633 Tel 55 115185 9833 Tel 888 259 0187 BD Biosciences Life Science Research Tel 81 35413 8251 Tel 525 237 1200 Tel 877 232 8995 Fax 858 812 8888 Fax 65 8601 590 Fax 55 11 5185 9834 Fax 905 542 9391 Tel 32 53 720211 Tel 49 6221 305 521 Fax 81 3 5413 8155 Fax 525 237 1288 canada bd com Fax 32 53 720 450 Fax 49 6221 305 531 MONOLIGHT 3096 Microplate Luminometer Operating Manual BD Biosciences www bdbiosciences com Clontech Immunocytometry Systems Discovery Labware For research use only Not for use in diagnostic or therapeutic procedures Not for Resale Canada Tel 888 259 0187 Fax 905 542 9391 canada bd com Brazil Tel 55 11 5185 9833 Fax 55 11 5185 9834 Asia Pacific Tel 65 8610 633 Fax 65 8601 590 PharMingen Transduction Laboratories E gt B D BD Biosciences United States Tel 877 232 8995 Fax 858 812 8888 Mexico Tel 525 237 1200 Fax 525 237 1288 Japan Tel 81 3 5413 8251 Fax 81 3 5413 8155 Europe BD Biosciences Tel 32 53 720 211 Fax 32 53 720 450 Europe Life S
41. a Example of Measurement without Injectors Since the detector needs some time before reaching its operating temperature the results measured just after power on may differ slightly from those measured after the detector has reached its operating temperature Therefore we recommend that you e start the instrument at least half an hour before running the first measurement e leave the instrument turned on between individual measurements L Open the instrument door L1 Turn up the frame of the microplate support tray 1 Place the prepared microplate on the support tray so that it sits correctly in the fitting piece and the letters are in the back Well A1 should be in the upper right hand corner LJ Turn the frame down and push it shut until it snaps into place L Check that the microplate sits correctly L Close the instrument door and push it shut until it clicks into place L If you have not done so yet Load the Simplicity software See section 3 7 Then the Protocol Manager appears Microplate Luminometer Installation and Commissioning L In the field Protocol type select the protocol type Raw Data PC and click lt Create gt The Raw Data PC Parameters dialog box appears with several tabs for input of the meas urement parameters Figure 3 13 Timing tab In the ET Timing EF General ERS Excell Thu Corrent Raw Data PC Mote All delays are between the start of the respecte events Parameters
42. ake sure there is still enough space in the plastic tub for the escaping wash fluid LI Click the Wash button to open the Parameters for Priming Injectors dialog box 21 LI Select the injector Inj 1 or Inj 2 LJ Enter the volume stroke ml and the number of strokes for washing and priming Q The programmed procedure starts as soon as you click lt OK gt You may stop a started wash cycle any time by clicking the Stop Priming button 4 Microplate Luminometer Maintenance 8 7 Cleaning MONOLIGHT 3096 Injector Pumps and Tubings The MONOLIGHT 3096 is equipped with a syringe injector sys tem and Teflon tubings All materials coming into contact with reagents are chemically resistant Daily routine cleaning procedure Use deionized or distilled to remove the reagents that are still inside the injection system Place the priming container on the microplate loading area Start your measurement protocol then initialize the injector s at the end of a measurement this step may be skipped 1 Click on the lt Priming gt button and select the injector s you want to clean 2 Select a volume of 500 ul and 10 injection cycles strokes 3 Click lt OK gt to start the wash process 8 8 Basic Cleaning Before or After Long Breaks Acid Base Cleaning Procedure This basic cleaning must be done before or after the injector has not been in operation for a longer period of time To keep the fluids for about 10 minutes in
43. ared from the screen and all tool buttons are active again You may keep the selected area or de fine a new one as describe above b When using anew measurement protocol To load a new protocol you must to exit the measurement menu via File Exit The program changes to the Protocol Manager and you can select another measurement protocol and start with lt Run gt To transfer the measured data to Excel click the button Ex cel is started provided it is installed on your PC and the measured data are imported into an Excel spreadsheet 49 Microplate Luminometer Figure 4 5 Presentation of the measured results in an Excel spreadsheet 50 Printout Copy Structure of the Software w Microsoft Excel Tabellel Datei Bearbeiten Ansicht Einfugen Format Extras Daten Fenster 7 x Oe BAY sBBSlo o BSl A lM Arial 10 F x u 228 98 m m o A O S lhlhCl it i E a a E 1 Background 0 Date 10 11 98 Time 16 23 00 2 Comment 3 RLUs Ad AQ AG Ad 4 Ds 133 123 233 145 r Jaf gt Zeichnen e 5 AutoFormen p OCAS gt 2 A EZRM tu oO i j jal g T m jms Bereit Click the Print button to print the measured data in a tabulated form If you have entered a comment it will be printed as well To set the print parameters select Print Setup on the File menu To view and edit the layout before printing select Pri
44. ay is accessible when the instrument is open and the frame is turned up If it is dirty clean it using a moist cloth and use cotton swabs to clean the corners If liquid has entered the instrument below the microplate tray push it into the measurement chamber provided you have al ready cleaned it and clean the bottom below the transport unit When using the injector unit option If liquid has entered the area below the service door either be cause the screw fittings at the injector head are not tight or due to leaky tubings clean this area using a moist cloth Before you open the service door disconnect the MONOLIGHT 3096 from the wall outlet and open the instrument door Microplate Luminometer Figure 8 1 Power supply unit on the instrument rear panel CAUTION Maintenance 8 2 Fuse Replacement MONOLIGHT 3096 The fuses on the instrument rear panel must be replaced when they are faulty or when changing the operating voltage see 8 3 The fuses are in a black fuse holder next to the power cord con nector see Figure 8 1 Power cord connector Power switch Recess for screwdriver Fuse holder Proceed as follows You need 1 small flat head screwdriver Spare fuses For operating voltage 230 V Fuse 160 mA slow blow For operating voltage 115 120 V Fuse 315 mA slow blow Use only UL approved fuses L Turn power switch to off 0 L Unplug the power cord from the wall outlet
45. cience Research Tel 49 6221 305 521 Fax 49 6221 305 531 Revised 6 19 01 Microplate Luminometer Contents 1 2 2 1 2 2 2 3 2 4 2 5 2 6 3 1 3 2 3 3 3 4 3 5 3 6 3 7 3 8 3 9 3 10 3 11 4 1 4 2 4 3 Contents Safety NS TuUCON Senn a aa 1 System Descriptio M rossarit dinini oa raa a aiaia 3 OVERVIEW occire 3 MONOLIGHT 3096 Microplate Luminometer ccccccceeeeeeeeseseees 6 2 2 1 Operating Components on the Front Panel ccecceseeeeeeeeeees T 2 2 2 Microplate Loading Compartment cccssccccceeeeesssessseeeeeeeeeeens 8 2 2 3 Measurement Chamber cccccccccceeccesssssneeeeeeeeeessssseeeeeeeeeeeesns 10 22 A LONCDELOC lO erna a a S 10 2 2 5 Background Measurement and Subtraction cccceeeeeeees 11 2 2 6 Injector Compartment behind the Service Doot 0008 11 2 2 Connections on the Rear Panel cece eeeesseeceeeeesssssseeeeeeeeeeens 14 INJECTOR UNIT CODUOMAN ssaa a a 15 MICODIa leS pcateseraesnh be hae E tenn uieeata dade T 16 PO barrett ads a eaves estan eel eu niga ae eee Se oe baa aaa ean Sanaeeas 17 SOMWA Oan re te tanaiaabva can eeaniaadaten toast nuns diluguadabupioastnungarearonavaianat 18 Installation and COMMISSIONING ccsssssseceeesssssseeeeeesssssceeeeesnesseeeees 19 OW CP VICW remire O 19 SCP Sle nen tenn mete ne ee an a ee eee eee eee 20 SPACE REUNEA seron tananasadanderseenassaanaaaeans 20 MIND G UNO ee
46. d Thiz unregistered version of the Fast Kinetics Photon Counter protocol will run for 30 days after installation Please register before that time to enable unlimited use Days left T To obtain the password to enable the software please contact your distributor Have the System ID handy when vou contact the distributor System ID 19842 Please enter your registration information here Mame Company Password Register How Cancel If you did not enter a correct password after 30 days you cannot call a protocol anymore The Registration Form dialog box appears again and you either have to enter your registration password or quit this dialog box with lt Cancel gt Microplate Luminometer Installation and Commissioning Registration To get your registration password contact BD Biosciences Pharmingen Technical Services 1 800 825 5832 lf the Registration Form dialog box appears when you call a protocol enter your name the company name and your password Then click lt Register Now gt If your password entry is correct you can continue working with the software The Registration Form dialog box does not come up any more After registration you may uninstall the program using the command Uninstall and install an updated version without having to enter a registration password 33 Microplate Luminometer 34 CAUTION Installation and Commissioning 3 11 First Measurement with Raw Dat
47. dialog box M Use injection 1 Volurnie fio ul uu Use dual measurement Injection 1 to measurement delay 1 23 amp me Measurement duration d First the Timing tab appears _1 When only one injector is used select either Use injection 1 or Use injection 2 if your system includes the Injector Unit L Enter required measuring time per well The default value is 1 second L Select the General tab and check the microplate type you are using 96 wells LI Select Take background at start of plate measurement if you want to run a background measurement prior to the sam ple measurement 35 Microplate Luminometer Figure 3 14 General tab in the Raw data PC Parameters dialog box 36 Installation and Commissioning Raw Data PC Parameters Raw Data PC Ea pty Titing E General EE excetrn Comment Microplate size C 394 wells Take background at start of plate measurement cara Ll The EXCEL TM tab is not needed for the first measure ments LI Select the Comment tab if you want to enter a comment for the measurement protocol It appears after the protocol has been stored in the Protocol Manager next to the measure ment protocol list when the respective measurement protocol is selected Li Click lt Save As gt and enter a file name in the following dialog box It must differ by at least one character from the file name of the selected protocol type L
48. e Window gt Show Table sults El Graphical presentation of re Window gt Show Kinetics sults Graph Graphical presentation of indi Window gt Graph wells vidual wells Zoom function Window gt Zoom Windows arranged horizontally Window gt Tile Horizontal Windows arranged vertically Window gt Tile Vertical AE Initializing the injectors Washing Priming the injector in tubings Pal Stopping the washing priming process Options without buttons Options gt Autosave If this item is active indicated by a W the measured data are saved automatically upon completion of the measurement The measured data are saved according to the measurement method used to consecutively numbered files and cleared from the screen Thus you can start another measurement using either the selected sample area or defining a new area immediately File gt Exit Exit the measurement menu and return to the Protocol Manager File names of autosave files Raw Data RD1 TXT RD2 TXT RDQMxxx TXT Slow Kinetics SK1 TXT SK2 TXT SKXXx TXT Fast Kinetics FK1 TXT FK2 TXT FAXxx TXT 53 Microplate Luminometer 54 Creating a new measurement protocol Editing an existing measurement protocol Figure 5 1 Timing tab in the Raw Data PC Parameters dialog box Raw Data Raw Data 5 1 Overview The function Raw Data allows you to run measurements with
49. e e g cleaning DLR_Assay Proceed as follows 1 Wash with deionized or distilled water 1 1 Use deionized or distilled water to remove the reagents remaining in the injection system 1 2 Place the priming container on the microplate loading area 1 3 Start the cleaning protocol and initialize the injectors 1 4 Click the lt Priming gt button and select the injector s you want to clean 1 5 Select a volume of 150 ul and 20 injection cycles strokes 1 6 Click lt OK gt to start the wash process 1 7 Repeat steps 1 1 1 6 2 Cleaning with Ethanol 2 1 Fill the reagent bottles and injector s with 70 Ethanol 2 2 Remove front cover of injector unit to watch the syringe movement 2 3 Click the lt Priming gt button and select the injector s you want to clean 2 4 Select a volume of 150 ul and 20 injection cycles strokes 2 5 Click lt OK gt to start the wash process 2 6 Repeat steps 2 3 2 5 3 Re washing with deionized or distilled water 3 1 Wash the syringe with deionized or distilled water using the priming routine steps 1 1 1 7 20 cycles strokes with 150 ul each 3 2 Repeat the wash process with deionized or distilled water to remove all traces of ethanol 101 Microplate Luminometer Maintenance 8 10 Establishing Tubing Connections If a tubing connection at the pump head or at the injector head is faulty or to replace an injector tubing you must provide a new tubing connection To
50. e pump head left connection OUT Proceed in the same manner with injector tubing 2 Pass the injector tubings at the MONOLIGHT 3096 over the rubber foam piece and at the injector unit through the openings on the side Figure 3 4 MONOLIGHT 3096 Injector head Opening for injector tubings with connected to MONOLIGHT 3096 to reagent bottles injector unit both devices open Injector pump 2 Injector pumps Tray for reagent bottles LI Cut the tubings connected to the IN ports right of the pumps to the length you need and connect them to your reagent bot tles Make sure the tubings are long enough to pass them through the openings on top of injector unit and to the bottom of the reagent bottles standing on the tray attached to the front of the injector unit LI Close the injector unit again by inserting the locking door ver tically from above Pass both slots on the bottom side of the door through the openings at the bottom of the injector unit the two slots on top of the injector unit through the openings on the door LI Close the service door at the MONOLIGHT 3096 very carefully to prevent any light from entering the measurement chamber 24 Microplate Luminometer Figure 3 5 Rear panel of injector unit CAUTION Installation and Commissioning LI The tubings leading to the injector unit must be installed at the foreseen place rubber foam
51. e red lt Stop gt button when the syringe has reached the extended position and let the syringe stay in this posi tion for about 10 minutes Wash the syringe with deionized or distilled water using the priming routine 20 cycles strokes with 150 ul each 2 Cleaning Step Cleaning with 0 1N HCL Fill the syringe with 0 1N HCL and leave this solution for about 10 minutes in the syringe same as for cleaning with NaOH steps 2 2 2 6 Start a measurement by clicking the green lt Start gt button click the red lt Stop gt button when the syringe has reached the extended position and let the syringe stay in this posi tion for about 10 minutes Then wash the syringe with deionized or distilled water using the priming routine 20 cycles Strokes with 150 ul each Microplate Luminometer Maintenance 8 9 Promega Reagents Cleaning Procedure One of the Promega Dual Luciferase Assay reagents the Stop amp Glo Reagent has a slight affinity to plastic materials To avoid cross contamination when changing the injector tubings we recommend the follow the cleaning procedure outlined below To keep the fluids in the syringes for about 30 minutes move the syringe pistons to the extended position and hold them there To do this you must use a cleaning protocol Therefore create your own cleaning protocol from the protocol you are using for measurement set the injector volumes as 150 ul and then save the protocol under another nam
52. easured L Mark the well group you want with the mouse and press the Ics button to confirm your selection The selected wells are presented white and the measurement value table is dis played according to the selected wells with data points see Figure 6 9 Slow Kinetics PC Untitled Data Table File Edit Options wiew Measure Window Help Ole saa E eles SIO Se ofe alm eel 5 Background fo Date m 23055 Measurement duration 1 0 Mo Time pesem Fress the go button or space bar or enter to measure Please note Before running a measurement if desired select the autosave mode Select this function on the Options menu Microplate Luminometer Figure 6 10 Measurement menu Slow Kinetics with tabulated result display Corrected interval times Measurement is finished Slow Kinetics Procedure without injectors LI Insert microplate correctly into the luminometer and close cover LJ Run background measurement if necessary Zech LI Select the wells to be measured h Q Start measurement LI Data point 1 all selected samples are measured in suc cession and the measured values are displayed on the screen If you have selected too short intervals the system corrects the intervals using the actually required time LI Then the measurements at data point 2 are performed and the results displayed All following measurements are per formed in the same manner Q The measurement is
53. ection of workbook Click on lt gt to dis play the browser The subdirectory Sim plicity workbook is defaulted At cell The measured data are read starting at this cell of the Excel spreadsheets Excel TM macro Entry of an Excel macro Comment tab The comment entered here appears after the protocol has been saved in the Protocol Manager next to the measurement proto col list when the respective measurement protocol is selected Slow Kinetics PC Parameters Slow Kinetics PC ED Plate Timing ETY ell Timing E General EE Excell Tia E Comment The following buttons appear on all tabs of the entire measure ment protocol is selected to define a new file name This is necessary in order to create a new measurement protocol or to save an existing measurement protocol under a new name Clicking on this but ton opens the Save As dialog box Enter a file name which differs by at least one character from the file name of the se lected protocol type As soon as the protocol has been saved the program returns to the Protocol Manager and shows the name of the new measurement protocols in the field Runable protocols Microplate Luminometer lt Save gt lt Cancel gt Figure 6 8 Measurement menu Slow Kinetics with sample matrix and table of measured values Slow Kinetics Saves the parameters in the existing measurement protocol file After you have saved the protocol the program retu
54. ector inj 1 or inj 2 by clicking on the respective box Then enter the injector parameters Please note Injector 1 injects into the well before the meas urement position injector 2 in the measurement position Volume Enter the injection volume in ul via keyboard or by clicking on the arrow buttons Injection 1 to measurement delay Enter the delay time be tween the injection of injector 1 and the start of the measure ment in seconds The hardware dependent minimum value of 2 05 seconds is defaulted and can be increased Injection 2 to measurement delay Enter the delay time be tween the injection of injector 2 and the start of the measure ment in seconds The minimum value of 0 seconds is defaulted and can be increased Fast Kinetics PC Parameters Fast Kinetics PC pty Timing EF General l EE Excell Th l E Comment Note All delays are between the start of the respective events Volume fic ul pl l Use injection 2 Injection 1 bo measurement delay i 2a amp 2 H lt 3 Measurement duratior fio z Ze a 79 Microplate Luminometer Fast Kinetics When using two injector Select both injectors The following dialog box appears Figure 7 3 Fast Kinetics PC Parameters Fast Kinetics PC Well Timing tab C3 Timing General EE Excon E Comment Selection of Note All delays are between the start of the respective events injector 1 and 2 M Use injection 1 Volume i0 Su Volume fic ul
55. for example priming will be discussed at the end of this section All important functions in the measurement menu can be se lected via tool buttons Buttons or menu items that cannot be selected are dimmed Some buttons are dimmed gray for ex ample when a measurement is running or no contact has been established with the luminometer or the data of the last meas urement have not yet been saved In all these cases a meas urement cannot be started Microplate Luminometer Structure of the Software Autosave Function Upon selection of this function the results are automatically saved to files The file names comprise an abbreviation of the measurement method RD SK and FK and a consecutive number This ensures fast storage of the measured data ac cording to the selected methods Before starting a measurement first select the Autosave func tion on the Options menu W active It will remain active until you deselect it again Selection of the sample area to be measured see also page 39 All wells of the microplate As default all wells of a microplate are selected for the following measurement When you click the Start button Ej the entire mi croplate is measured Selecting partial areas If you want to measure one segment of the microplate click the cs button mark the desired area with the cursor Then click the Ics button again to confirm the selection and to exit the select mode see section 4 4 The we
56. hat EXCEL 7 0 is in stalled on your PC and running Figure 7 5 Fast Kinetics PC Parameters Fast Kinetics PC 81 Microplate Luminometer 82 Figure 7 6 Input box for a comment on the measurement protocol lt Save As gt Fast Kinetics Auto send to Upon completion of the measurement the Excel macro is automatically sent to the Ex cel program and executed there Workbook Selection of workbook Click on lt gt to dis play the browser The subdirectory Sim plicity workbook is defaulted At cell The measured data are read starting at this cell of the Excel spreadsheets Excel TM macro Entry of an Excel macro Comment tab The comment entered here appears after the protocol has been saved in the Protocol Manager next to the measurement proto col list when the respective measurement protocol is selected Fast Kinetics PC Parameters Fast Kinetics PC ae Timing EE General EE Excell Th E Comment rea The following buttons appear on all tabs of the entire measure ment protocol is selected to define a new file name This is necessary in order to create a new measurement protocol or to save an existing measurement protocol under a new name Clicking on this but ton opens the Save As dialog box Enter a file name which differs by at least one character from the file name of the se lected protocol type As soon as the protocol has been saved the program re
57. he available measurement protocols are listed in alphabetical order They are based on a selected protocol type and include the parameters defined by the user The following buttons are available to edit measurement protocols lt Run gt Opens the measurement menus to start a meas urement with the selected protocol lt Edit gt Shows the selected measurement protocol and lets you change it lt Delete gt Deletes the selected protocol 4 2 2 Instrument Tab On this page enter and view the instrument parameters and ini tialize the instrument Only the buttons lt Instrument options gt lt Check instrument gt and lt Take Background gt are important for the user lt Instrument options gt takes you to the Instrument Options dialog box to define the instrument communication port see section 3 9 Instrument Configuration Further Protocol Manager Options lt Retrieve Data gt Load and display stored measurement data lt Options gt Select subdirectory to which the measured data are to be saved lt Exit gt Exit program 43 Microplate Luminometer Structure of the Software 4 3 Parameter Input Select a protocol type and click lt Create gt or select a measure ment protocol and click lt Edit gt to display the respective meas urement protocol for parameter input Figure 4 3 The title bar shows the file name of the protocol This dialog box comprises several tabs To open a tab simply click on it
58. he luminometer The LED s from right to left Microplate Luminometer is turned on and ready for operation Measurement is running Do not open instrument door Data transfer between PC and luminometer Instrument door The instrument door for loading microplates is on the left in strument side It is opened and closed by hand When closing the door make sure to push it into the spring lock An audible click indicates that the instrument door is closed light tight If it is not closed correctly a warning will appear on the display For correct measurements the instrument door and the exon service door must be properly closed so that no outside light can enter the instrument Figure 2 3 MONOLIGHT 3096 Microplate Injector head and Injector tubings View with instrument loading area measurement head door and service door open ia Sot 74 i hr L A Wey AL Py r L 1 Fa Ea Le wy ho gr Ps ar Van RA al T NY y ch yu fy i L PMY ar KY har Gr Gd eh ey AN Ph ey Ph ey Ay AQ Lor Lo 7 h a Lor wr L Injector 2 on Lo Le a h Injector 1 Fhe wa uy oe d h py rH rh r H AA h g TK p7 T is AN a a a a a T a Lr m o o m ar g m Gr g Velcro fastening Microplate Luminometer Figure 2 4 Microplate transport unit with raised holding frame
59. ild household cleaner e g detergent but never scrubbing powder If liquid gets inside the instrument turn the pump off open the locking door and clean the dirty parts Determine the cause of the leak screw fitting not tight or pump defective and take the appropriate countermeasures tighten or replace the fitting screw see section 8 10 97 Microplate Luminometer 98 Maintenance 8 6 Priming Washing Cleaning the Injector Tubings Washing and priming of the injector tubings are controlled via the Simplicity PC software To clean the injector tubing system you should rinse it regularly with distilled or deionized water Further recommendations for cleaning see section 8 8 For measurement injections you must first prime the tubing so that the full volume is injected with the first shot Proceed as follows to wash or prime the tubing systems 1 Initialize the injector unit LI Select any measurement protocol that contains a measure ment programmed with the respective injectors click lt Run gt in the Protocol Manager LI Click the Initialize button in the measurement menu ick LJ This opens a window prompting you to place the plastic tub in the microplate loading compartment instead of a microplate to collect any escaping liquid LI As soon as you have positioned the plastic tub click lt OK gt The end of the initialization is indicated in another window 2 Washing and Priming the Injector Tubings LI M
60. ill see a corresponding message Simplicity 1 0 Alpha 4 Installation Ready to Install You are now ready to install Simplicity 1 0 Alpha 8 Protocols created prior to 1 0 alpha 7 will not work with this version of the software Please delete them then create your protocols again Fress the Nest button to begin the installation or the Back button to reenter the installation information Cancel 27 Microplate Luminometer 28 Figure 3 8 Simplicity Protocol Manager Installation and Commissioning 3 8 Program Start To start the Simplicity PC program click the Windows 95 lt Start gt button then select Programs and Simplicity PC Simplicity PC is loaded and the Protocol Manager displayed with two tabs see Figure 3 8 Simplicity Protocol Manager F Protocols lnetumert Protocol types Retneve Data Quick Measurement Quick Measurement iza Slow Kinetics Options fast and simple protocol to take measurements El Runnable protocols Cut off Assay Example Performs a cut off assay on Slow Kinetics 2 the first kwo columns of data using Excel 97 The program is configured such that the instrument is initialized only upon selection of the measurement menu lt Run gt In this manner you can first define the communication port used You can change this presetting in the Instrument Options dialog box so that the initialization takes place when the program is started
61. iming tab in the Fast Kinetics PC Parameters dialog box 78 7 1 Overview In the measurement mode Fast Kinetics the trend of the brief light emission of samples is measured The individual samples are measured in succession and the measured results are dis played in a table The data can be presented as a graph or ina table or imported into an EXCEL spreadsheet for further proc essing 7 2 Measurement Protocol In the Protocol Manager select the protocol type Fast Kinetics and click lt Create gt In the Protocol Manager select an existing measurement pro tocol and click lt Edit gt The Fast Kinetics Parameters dialog box with several tabs is displayed and you can enter the measurement parameters Figure 7 1 Fast Kinetics PC Parameters Fast Kinetics PC ELJ Timing General EE Excell Thal E Cormmert Hote All delays are between the start of the respective events Use injection 1 Use injection 2 Measurement duration Microplate Luminometer Fast Kinetics Well Timing tab without injectors Measurement duration Enter the measuring time per well The default value is 1 second Well Timing tab with injectors Except for the measurement time enter the parameters for the individual injections here When using one injector Injector 1 Injector 2 ui Microplate p gt Or Figure 7 2 Timing tab Selection of injector 1 Select the inj
62. ing the Injector Unit OUtSICe eee eeeeesssseeeeeeeseees 97 Priming Washing Cleaning the Injector TUDINGS cceeeeeeeeees 98 Cleaning MONOLIGHT 3096 Injector Pumps and Tubings 99 Basic Cleaning Before or After Long BreakS ccccccsscceeeeeeessneeees 99 Promega Reagents Cleaning Procedure cccsccccceeessssseeeeeeeseees 101 Establishing Tubing CONNECTIONS cccccccceeessssseeeeeeessseeeeeeesssees 102 REDIECING INJOClON TIP Snesere TEA R 104 Replacing Pump SYSINGES ccccccsssscecceesesssseeeeeeesssseeeeeeessssteeeeeeesseees 106 Replacing Fuses in the Injector UNit cc ceeeeeessssseeeeeeeeseeeseaes 107 Changing the Operating System Voltage in the Injector Unit 109 TecHMEaAl Dala unimini 110 Microplate Luminometer Typographical Conventions Typographical Conventions To make it easy for you to work with this manual and to use the software we will use the following typographical conventions throughout this manual Button labels are printed in bold typeface inside angular brackets Example lt OK gt lt Start Quick Measurement gt Menu and option titles are printed in bold typeface inside square brackets Example File Options Actions are symbolized by WU Enumerations are symbolized by i Microplate Luminometer 1 CAUTION Safety Instructions Safety Instructions The MONOLIGHT 3096 as well as the injector unit
63. ing with injectors special devices for injector tips and tubings In connection with the highly precise injector unit measurements with up to two injectors can be performed Operation and control of the MONOLIGHT 3096 Microplate Lu minometer the injectors as well as evaluation of the measured results take place via the Windows software Simplicity Photon Counter abbreviated Simplicity PC which was specifically de signed for this field of application It is based on MS EXCEL and features clearly structured and intuitive user guidance Three measurement modes based on predefined measure ment protocols are available Raw Data Quick measurement of a microplate Slow Kinetics Kinetics measurement for longer light reactions For each data point all Samples are measured in succession Fast Kinetics Kinetics measurement for short light reactions The trend of the light emis sion of the 1 sample then the 2 sample etc is measured The software is designed such that further measurement proto cols can be integrated This can be done by the user via EXCEL macros in the Simplicity PC software The MONOLIGHT 3096 Injector Unit works with the same PC software Corresponding to the current instrument configuration additional program functions are enabled when injectors are connected Microplate Luminometer System Description Fields of application The MONOLIGHT 3096 Microplate Luminometer can be used for these and other applicat
64. injector unit optional to the 9 pin socket of the MONOLIGHT 3096 and to the 15 pin socket on the injector unit Connect the instrument to wall socket via the supplied power cable Line voltage and operating voltage must always match CAUTION Observe a minimum distance of 10 cm between the back of CAUTION the instrument and the wall to allow air circulation for the cooling element Voltage change see section 8 3 Fuse replacement see section 8 2 14 Microplate Luminometer Figure 2 8 MONOLIGHT 3096 with connected injector unit both devices open System Description 2 3 Injector Unit optional The MONOLIGHT 3096 injector unit consists of two highly pre cise Cavro pumps and the control unit which is integrated in the housing The injector unit is programmed such that the left pump is con nected to the left injector tip in the MONOLIGHT 3096 In the Simplicity PC software the left injector is Injector 1 Microplate Injector head Injector Injector unit loading area tubings qer Injector 2 Injector 1 Tray for reagent bottles The right pump is connected to the right injector tip in the MONOLIGHT 3096 In the Simplicity PC software the right injector is injector 2 The injector unit is closed by a plastic cover on the front which can be taken off to connect the tubing and to service the injector unit see section 8 Lift the cover up vertically from above Pass the injector tubings left
65. ions e Reporter gene assays e Immunoassays e ATP assays DNA and protein assays PCR quantification assays Figure 2 1 Components of the measuring system a MONOLIGHT 3096 Luminometer b Injector unit c PC or laptop d Printer Microplate Luminometer CAUTION Figure 2 2 MONOLIGHT 3096 Microplate Luminometer with injector unit front view System Description 2 2 MONOLIGHT 3096 Microplate Luminometer The MONOLIGHT 3096 Microplate Luminometer is a compact flat desktop unit with small footprint due to its small size it can be set up on any lab workplace The MONOLIGHT 3096 can work without or with up to 2 injectors Simply connect the MONOLIGHT 3096 injector unit to the MONOLIGHT 3096 To obtain reliable consistent results keep the following in mind Do not expose instrument to direct sunlight or heavy temperature fluctuations Set instrument up in dry rooms Keep microplate loading compartment free of dust and dirt Clean spilled reagents inside the instrument immediately using a clean and dry cloth Open instrument door only for loading or cleaning to keep the inside dust free Open service door next to the instrument door only after having disconnected the instrument from main and when the instrument door is open Microplate Luminometer System Description 2 2 1 Operating Components on the Front Panel Four LED s inform you at one glance about the current status of t
66. ions The instruments are live and improper handling may cause damage Before opening the instruments disconnect the power sup ply Switch off the instruments before pulling the power cord Spare fuses must match the values specified by the instru ment manufacturer The fuses must not be short circuited or tampered with 10 All instruments supplied and all additional devices must be 11 12 grounded Use three pole grounded plugs If you can see that the units have become unsafe to use switch them off and disconnect them from the power supply If liquid gets inside the instruments pull the power cord Clean the unit or have it cleaned by an authorized service center The tests and maintenance work recommended by the manu facturer should be performed to make sure that the operator re main safe and that the instrument continues to function correctly Any service and maintenance work not described in the operat ing manual must be performed by authorized service engineers Microplate Luminometer 2 Highly sensitive light detector Low crosstalk Mechanical construction System Description System Description 2 1 Overview The MONOLIGHT 3096 Microplate Luminometer has been de signed for the detection of chemi and bioluminescence and for all measurements of glow and flash luminescence on mi croplates i e for reactions where the light remain nearly con stant over a longer period of time as well as
67. lations become visible average peak etc You may also increase the graphics section by moving the hori zontal bar up so far that the entire data section is hidden in ie Co th Ca fe LT in i i ni a Microplate Luminometer The graph window Fast Kinetics The curve trend of the measured values is presented in the graph window default setting automatic scaling The well po sitions and associated colors are listed in the graphics legend next to the diagram Options in the graph window Smoothing Log scale RLU range Automatic Manual Smoothing the measured values Logarithmic scaling of the measured val ues Selection of scaling Default setting is automatic scaling of the measured RLU values comprising the top and bottom value of each coordinate Manual setting option for the display area Upon selection of this item you may enter the start min and end max of the RLU ranges When you click Manual once more the entered range can be applied to the graphics 89 Microplate Luminometer 8 90 Maintenance Maintenance 8 1 Cleaning the MONOLIGHT 3096 The outside of the luminometer is protected by a rugged wash able varnish If it is dirty or dusty clean it using a moist cloth If necessary use a mild cleaner e g detergent but never any scouring powder Keep the microplate transport unit and frame clean The spring loaded black microplate support tr
68. ld by Velcro fastening The plate covers the instrument area with the injector tubings the injector tip holder and the measurement head see Figure 2 5 Caution Since the photomultiplier is exposed to incident light when the service door is opened you must disconnect the in strument from main before opening it to rule out damage to the photomultiplier For measurement the service door and the in strument door must be closed properly Never operate the in strument with the service door removed e Before opening the locking plate next to the instrument door disconnect the instrument from wall socket and make sure the instrument door is open e First open the 2 fastening screws of the locking plate and hold the plate at the screwed side and take it off Otherwise the locking plate may get damaged CAUTION 11 Microplate Luminometer System Description gues Microplat Injector head and Injector tubi icroplate njector head an njector tubings MONOLIGHT 3096 loading compartment measurement head View with open instrument door and service door oo 20 Injector 2 fou Ly _ Injector 1 Foam for cable feed through Lor h s d L PE SEP L d L N a M eh eh ey aa BW e Ld a h L r Ld Velcro fastening Insrument Door Service Door The injectors are arranged as shown in Figure 2 6 The mic
69. lls selected for measurement are depicted white on the screen the others shaded gray Background measurement Before starting a measurement you can run a background measurement by pressing the BG button The result is dis played in the Background field Select Subtract if you want to subtract the background of the individual sample values Measurement start Measurement of the microplate or the selected area starts after pressing the Start button green signal light when the instru ment door is closed The red LED lights up on the instrument The Start button is dimmed when a measurement is running when there is no contact with the luminometer or when the data of the last measurement have not yet been saved In all these cases a measurement cannot be started 47 Microplate Luminometer Structure of the Software 48 lf the instrument door is not closed a warning appears inform ing you about the current measurement mode Raw Data PC E Door open error ERR_COOR_OPEN Close the instrument door and click lt Repeat gt or lt OK gt Then the measurement starts Measurement procedure The measurement procedure varies depending on the selected measurement method Measurement end The measurement is over when all selected wells have been measured In the autosave mode the measured results are stored automatically and the measured values cleared from the screen If the autosave mode is not active the measu
70. minimum preparations The measurement starts as soon as you have entered the measurement parameters and defined the wells to be measured Then the measured data are transferred to the PC every 0 2 seconds and the averaged results are dis played on the screen in a sample matrix The measured results can then be imported into EXCEL for further processing 5 2 Measurement Protocol In the Protocol Manager select the protocol type Raw Data and click lt Create gt In the Protocol Manager select an existing measurement pro tocol and click lt Edit gt The Raw Data PC Parameters dialog box appears with the file name of the measurement protocol being displayed on the title bar This dialog box comprises several tabs for input of the measurement parameters Figure 5 1 Raw Data PC Parameters Raw Data PC x ET Timing E General l EE excell Thi E Comment l Note All delays are between the start of the respective events Use injection 1 Use injection 2 Use dual measurement Measurement duration Microplate Luminometer When using one injector Injector 1 Injector 2 fF fF Microplate Or Figure 5 2 Timing tab Selec tion of injector 1 Raw Data Timing tab without injectors Measurement duration Enter the measurement time per well The default value is 1 second Timing tab with injectors Except for the measurement time you enter the parameters for
71. mum value of 2 05 seconds is defaulted and can be increased Injection 2 to measurement delay Enter the delay time be tween the injection of injector 2 and the start of the measure ment in seconds The minimum value of 0 seconds is defaulted and can be increased Figure 6 3 Well Timing tab Selection of injector 7 When using two injector Select both injectors The following dialog box appears Figure 6 4 Slow Kinetics PC Parameters Slow Kinetics PC Well Timing tab ET Plate Timing ET Well Timing Selection of injector 1 and 2 ANAE d wl w gt d l cap EA dJ al me Es 67 Microplate Luminometer Slow Kinetics Volume Enter the injection volumes in ul via keyboard or by clicking on the arrow buttons Injector 1 Injector Injection 1 to injection 2 delay Delay time between the injec N tions of injector 1 and 2 The hardware dependent minimum el value of 2 05 seconds is defaulted and can be increased tp Injection 2 to measurement delay Enter the delay time be Lt tween the injection of injector 2 and the start of the measure Microplate p ment in seconds 0 second is defaulted and can be increased tm 1 Example Programming a measurement with 2 injections bt r Pall P _ pre injection injection M measurement tp delay between pre injection and injection tu delay between injection and start of measurement Ty duration of measurement
72. n be ordered separately Luminometer packing material is Cat No 551484 and the Injector Unit packing material is Cat No is 551485 Unpack all units and accessories and ensure the shipment is complete and shows no sign of transport damage The careful packing usually rules out transport damages Should the instrument or instrument parts be damaged please inform the shipping agent or the BD Biosciences Pharmingen immediately Please be careful when unpacking the instrument and removing the foamed inserts Microplate Luminometer Installation and Commissioning 3 5 Connecting the Instruments LI Carefully take the Microplate Luminometer out of the cardboard box and place it on your lab bench L Open the instrument door Li Using a flat head screwdriver unscrew the red transport securing screw which holds the microplate transport unit to the right next to the microplate frame 1 Keep the screw and use it again to transport the instrument To the left of the sample compartment there is a threaded hole in which the screw can be stored by screwing it in CAUTION Save screw and install it again when transporting the instrument Figure 3 7 Microplate loading compartment with transport securing screw Borehole for transport screw Storage of transport screw Li Check if the wall socket voltage matches the operating voltage labeled on the instrument rear panel label and position of the white pin on the fuse holder
73. n the instrument rear panel Maintenance 8 13 Replacing Fuses in the Injector Unit The fuses on the instrument rear panel must be replaced when they are faulty or when changing the operating voltage see 0 The fuses are stored in a black fuse holder next to the main plug see Figure 8 5 F eeCcoscase F O F sS C E E ve Fuse holder Recess for screwdriver Proceed as follows You need 1 small flathead screwdriver Spare fuses For operating voltage 230 V Fuse 160 mA slow blow For operating voltage 115 V Fuse 315 mA slow blow Use only UL approved fuses LJ Turn main switch to off position 0 LJ Disconnect power cord from wall outlet LJ Insert a screwdriver or a similar tool into the recess on the right side of the fuse holder push slightly and take the fuse holder out The safety holder includes one fuse each for op erating voltage 115 V and 230 V LI Remove the faulty fuse Faulty fuses can typically be detected by a broken fuse wire in the glass envelope 107 Microplate Luminometer 108 Maintenance LI Insert new fuse s and observe the required current and fuse ratings Otherwise the operating safety cannot be guaranteed and the instrument will lose its UL Underwriters Laborato ries approval LI Install fuse holder correctly so that proper operating voltage is indicated Function check LI Connect power cord to wall outlet LJ T
74. nt Preview on the File menu Copy measured data to the clipboard Select the respective data with the cursor and click on this tool button You may then paste the copied data into another Windows program Microplate Luminometer Initialization Priming Structure of the Software 4 5 Special Functions when Using Injectors Prior to the first measurement after starting the programs you must initialize the injector unit by clicking the Initialize button Otherwise no measurement can be started Then you are prompted to place a plastic tub into the microplate loading com partment instead of a microplate to collect any spilled liquid As soon as you have placed the plastic tub click lt OK gt The end of the initialization process is again indicated in a window while the pumps perform several lifting motions Take the plastic tub out and insert the microplate to be meas ured then click lt OK gt in the message window Start the meas urement by clicking the green signal light Washing and priming of the injector tubings is possible only when the injector unit has been initialized Make sure you have placed an empty plastic tub in the microplate loading compart ment to collect spilled liquid Click on the Prime button to open the Parameters for Priming Injectors dialog box Select the injector Inj 1 or Inj 2 For each injector enter the volume strokes ml and the number of strokes The programmed priming process s
75. oint LI The measurement is finished when all predefined measure ments have been carried out Figure 7 9 85 Microplate Luminometer 86 Figure 7 9 Measurement menu Fast Kinetics with tabulated result display Fast Kinetics Procedure with injections LI Select the wells to be measured LI Place plastic tub in plate loading compartment s Po i ama C Q Initialize injectors LI Prime tubings if necessary Gt Insert microplate LI Insert microplate correctly in luminometer and close door B Q If necessary run background measurement 2 LI Select the wells to be measured E O Start measurement _ LI The preselected injections are performed and each sample is measured in succession according to the preselected measurement time The measured values are presented for each data point LI The measurement is finished when all predefined measure ments have been performed Figure 7 9 Fast Kinetics PC Untitled Data Table File Edit Options View Measure Window Help ce eka e ee ea ee jes Background a Date a ragr99 Measurement duration 1 0 m iie Time 032652 Integral 7 TOTE Kinetics 00 00 000 39450 39550 Microplate Luminometer Fast Kinetics 7 4 Result Display Graphical and tabulated display of the measured results e As needed you can call the window for graphical display of the measured data before during or after completion of the measurement
76. olumns of measurement protocol Eat Ate taoa ee Comment on highlighted ee protocol Exit program 4 2 1 Protocols Tab Protocol types The implemented protocol types are displayed in this field Each protocol type includes a group of parameters for a special measurement method such as the measurement time but with out any values The name of a protocol type therefore refers to the measurement method for which it is used for example Slow Kinetics for kinetics measurements with light emission over a longer period of time A protocol type always serves as template for a measurement protocol which includes certain pa rameters defined by the user Based on one protocol type you can set up aS many measurement protocols with different pa rameters desired To create a measurement protocol click lt Create gt i e select a protocol type as template enter the required parameters and save it under a new name The program stores measurement protocols automatically in the field Runable protocols and lists them in alphabetical order The name of a measurement 42 Microplate Luminometer Structure of the Software protocol must not be identical with the name of the protocol type but should refer to the measurement method You can enter a comment for each measurement protocol it appears in the field next to the measurement protocol list when the respective protocol has been selected see Figure 4 2 Runable protocols T
77. or after completion of the meas urement by clicking tel The graph display is on top of the table window Figure 6 11 BE Rees Ciri i TF fie it Gets jen Eee ake es olaj azie Atm Pe deel ee el io e To display the curve trend of one or several selected sam ples select the desired wells in the sample matrix with the cursor and click E e To view the graph and table window on the screen at the same time click or depending on the layout you prefer Figure 6 12 75 Microplate Luminometer Figure 6 12 Measurement menu Slow Kinetics with tabulated and graphical result display 76 Slow Kinetics e To open or close one display mode click the button tel or Ea The remaining window will fill half the screen e To maximize the window click or ual Further options for opening closing arranging maximizing mini mizing the size of graph and table windows Buttons in the upper right corner of the dialog box I Maximizing the windows to full size Ei Minimizing the windows to half the screen size Buttons in the upper left corner of the dialog box Opens a pulldown menu including the item Close to close the table window i F Opens a pulldown menu including the item Close to close the graph window Microplate Luminometer Figure 6 13 Graph window Slow Kinetics partial view Here you may pull the dividing bar up or down with the mouse
78. ples Microplate Luminometer Slow Kinetics e the time required for the movement of the microplate transport unit This is in turn dependent upon the position of the sam ples to be measured If the samples are located all over the microplate more time is needed than for an arrangement in one row or in one corner of the microplate at least 1 sec Please note for each data point interval all selected sam ples are measured in succession Calculation of actual times Total duration of experiment Duration of the interval x number of data points Duration of interval Delay times measurement time feed time x number of samples Total duration of experiment Total duration of measurement including movement of the mi croplate transport unit No of repeat plate measurement Number of data points Interval The program shows the interval period calculated from the two previous parameters During this time the defined samples should be measured once The minimum duration should be number of samples x 2 sec Parameter input The interval is defined by changing the parameters Total dura tion and No of repeat The program calculates the inputs in the dialog box according to the formula Interval Total duration No of repeat This demonstrates that the total duration must be increased when the interval is to be extended while the number of data points stays the same Please note Use in
79. red values remain displayed and can be stored later Measurement stop Press the Stop button red signal light to stop the measure ment The results measured up to now can be stored Comment To enter a comment for the measurement click the Comment button The text entered in the Measurement Comment dialog box is printed in the report and saved to the data file Save You can save the measured results under any name by clicking the Save button If you want to quit the measurement menu without saving the data or if you click the button for new measurement DI you will be prompted to save the data In the autosave mode each measurement is stored automati cally and the file name RD SK or FK is provided with a con secutive number Microplate Luminometer Structure of the Software New measurement Excel Transfer a When using the same measurement protocol In the autosave mode you can immediately start a new meas urement as soon as a measurement is finished You can keep the selected area or define a new area as describe above In the normal save mode the measurement is cleared for the next measurement when you click on the is button new measurement if the measured data of the last measurement have already been saved Otherwise you may continue only after you have either saved the data or confirmed that you do not want to save the data Clearing means that the measured data of the previous measurement are cle
80. rns to the Protocol Manager The changes are disregarded and the old values are still valid The program returns to the Protocol Manager 6 3 Measurement As soon as you have selected the measurement protocol type Slow Kinetics and you have clicked lt Run gt the program changes to the measurement menu Slow Kinetics Communi cation with the Microplate Luminometer is established Initial izing instrument appears on the status bar and the yellow LED lights up In the measurement menu Slow Kinetics the sample matrix is displayed first and below that a table for the measured values The table includes row by row the predefined data points for all samples on the microplate see Figure 6 8 Ameasurement of the entire plate can also be started immediately by pressing the Start button Slow Kinetics PC Untitled Data Table OF x File Edit Options View Measure Window Help He Sie Ee Pi Sil Sie Sle Tee ml Ey Background lo Date jm f29 99 Injection 1 volume 10 Injection 2 volume 10l Subtract Time 085220 E ca sontitGs m o doa dad 8 Js _ 1 Press the go button or space bar or enter to measure 71 Microplate Luminometer Figure 6 9 Measurement menu Slow Kinetics with selected well groups Measurement value table with prede fined intervals They are changed by the program upon completion of the first interval if they are too short 12 Slow Kinetics Selecting the wells to be m
81. roplate moves from left to right Injector 1 injects in the position before the measurement position injector 2 into the well in the measurement position see Figure 2 6 Figure 2 6 Injector 1 in Injector 2 in Position of injectors position before meas position i Microplate meas position above microplate Vin Measurement head You may take the injector head off to replace the tips and the in jector tubings see section 8 Maintenance 12 Microplate Luminometer System Description Closing the Service Door LI Close the service door carefully to ensure light cannot enter the measurement chamber LJ Put the tubing leading to the injector unit on the rubber foam so that the door is pressed into the foam and is closed light tight LI Align the service door exactly LI Then press the service door down firmly to close the Velcro fastening 13 Microplate Luminometer System Description 2 2 Connections on the Rear Panel The Luminometer rear panel includes the connection ports the power switch the fuses and the cooling element Figure 2 7 Rear panel of MONOLIGHT 3096 Luminometer UUU E L Power switch Power cord Cooling Injector PC port Connector Element unit data port Connect the MONOLIGHT 3096 to the serial communication port of the PC using the supplied 9 pin data cable with two female connectors Using the supplied cable with two male connectors connect the MONOLIGHT 3096
82. rotocol Manager Figure 4 1 Structure of Simplicity PC program E3 Protocols Instrument l Protocol types Quick Measurement Slow Kinetics Runnable protocols Run Cut off Assay Example Slow Kinetics 2 Delete Create meas protocol Quick Heausurement Parameters Quick Measurement Save General ERE Exceli Thi E3 Comment l Take background at start of plate measurement Structure of the Software Simplicity Protocol Manager Retieve Data Options Quick Measurement iz a fast and simple protocol to take measurements ia Performs a cut off assay on the first two columns of data using Excel 97 Measurement menu 4 Microplate Luminometer Structure of the Software 4 2 The Protocol Manager After startup of the software the Protocol Manager appears and you can create and edit protocols and start measurements Creating instrument parameters Figure 4 2 Simpli CI ty P r otocol Simplicity Protocol Manager Manager Protocols lt lt netument Retrieve Protocol types Retrieve Data measured Quick Measurement Quick Measurementiga data low Kinetics fast and simple protocol to Options Protocol take measurements rectory for El measured Runnable protocols data U ted Run Cut off Assay Example Performs a cut off assay on ser creale Slow Kinetics 2 the first two c
83. se a deceeten satan tea teasenes 20 Connecting the INStrUMENHS ccccccccessssseeeeeesssseceeeessssseeeeeeeessseees 21 Connecting the Injector UNII ccssar 23 SOMWAlES STAN AUON sas ci sitsrcesiaatisaiennenolinctjeduestinianasdnawenendtantndddedvdeeocmaaandiaoiee 26 Program Staib calec yet ists anaes nauiac seer aa eteaanshlcesye 28 INSTFUMENT COMMOUATION lt 2scese cczcusssescoandecsanse doaseaesedetenmaeeseuadedeacsandazsegzeee 29 SoftWare REGIST ALON ia orcwectndctnerdsduriered ee ceawesea ubuseactatenes 32 First Measurement with Raw Datta u ccccccccccccecssssssssessseeeeseeeeeeeeeees 34 Structure Of the SOMWare icici enc Maids eee eae 40 Software Structure cece eeeeseeeeceeeeesssesseeeeeeeeeesssesseeeeeeeeeeesseesseseeeeeeeeeeesas 40 NG PrOlocol IVa ag CF siccaces roth a et eee ete nee seta 42 ALl PYOWCOIS TaD aie Aes 42 4 2 2 Instrument Tab cc cccecceeessseeeeeeeeeeeeeeeeeeeessssssssssssssssaaeees 43 4 2 3 Further Protocol Manager Options ccccccsseceeesssseeeeeeeeeeees 43 Parameter INDUL asnon a E A Seas anak 44 Microplate Luminometer Contents 4 4 4 5 4 6 oA 9 2 9 3 6 1 6 2 6 3 6 4 7 1 7 2 7 3 7 4 8 1 8 2 8 3 8 4 8 5 8 6 8 8 8 8 9 8 10 8 11 8 12 8 13 8 14 9 Measu urement Men onsen e 45 4 4 1 Structure of the Measurement Menu cccssceeeeesestteeeeeees 45 4 4 2 Working with the Measurement Menu ccccccccee
84. ssssesstteeeees 46 Special Functions when Using INjeCtors ceessseeeeeesssseeeeeeeseees 51 Software Functions at One Glance esesennnnnnnosnnnnnnoennnnnnneennnnnnnernennne 52 RAW Dalarna 54 OVE NIE Wa dalc eaatcia ae entice reader eae va ea manana etenarea Seacteaeae aia 54 Me easuremenl PLOlOCO vis iacasszeudavindsdaasernincsnsdentavananaessabaayaasderniacaasadaabonaans 54 MeasuremeMscrreo iana a a E lens 60 Slow Kinetics annnnnennnnnnnnnnnnnennnnnnnnnnrennnnnnnnnnnennnnnnnnnnnnn nn nnnnnnnnnnn ennen nnmnnn nnee 63 OV ChVIEW snoa a e es 63 Measurement Protocol ccccccccessssceeeeeeeeeeeeeeeeesssssssesssaaeeeeeeeeeeeeeeeees 64 WGEASUIEIN GI cisiscamsaeton aaa aa ee ee 71 RESUL DISIA y eucan e T eae 15 ASU INIT OU CS annaa a 78 OVC NIE W atereaicdinctoasactns ions ances E aeteeeeen eee 78 Measurement PrOlOCOknisamiornanu e a a 18 Meas Weme airian E 84 PRS SUE IIS DII oea sane annstaasaeradangutsaseseaeestnnanet 87 Mantena Ne e eee nee oP ae A A 90 Cleaning the MONOLIGHT 3096 ccccccssccccessssseeeeeeessseeeeeeesssseees 90 Fuse Replacement MONOLIGHT 3096 ccc ecceseeeeeeeseseeeeeens 91 Changing the Operating Voltage for MONOLIGHT 3096 93 Taking the Instrument out of Service for Transport e 96 servicing and Cleaning the Injector System 97 8 5 1 Regular Maintenance and Che cKG ccccccccsssceeeeeessseeeeeeeseees 97 8 5 2 Clean
85. t to initialize the Microplate Luminometer automatically any time the program is started Otherwise an initialization is per formed only after you select the measurement menu or click the lt Check instrument gt button on this tab Upon delivery of the software this item is not enabled d Take background at startup Select this item if you want to run a background measurement any time the program is started L Confirm your entries by clicking lt OK gt The Instrument Op tions dialog box is closed the Instrument tab is active Fig ure 3 9 LI Click lt Check instrument gt to establish communication be tween PC and luminometer If this is not possible an error message appears and you are prompted to check the con nections and the communication port parameters If an ini tialization is performed this is indicated on the status bar and by the yellow LED lighting up on the luminometer while the instrument modules are being checked LI Click lt Read configuration gt Then the current instrument configuration with or without injector s is read in from the Simplicity PC program You may carry out the following steps if required 1 To run a background measurement remove any light source from the measurement chamber close the loading cover and click lt Take Background gt While the background measure ment is on the red LED lights up on the luminometer The measured background value is displayed and remain stored
86. tarts as soon as you click lt OK gt When the priming process is finished you can start the measurement using the selected measurement protocol Stopping the Priming Process You may stop a started priming process any time by clicking the Stop Priming button SI Microplate Luminometer 52 Button Structure of the Software 4 6 Software Functions at One Glance The Simplicity software is a Windows 95 application and is therefore operated according to the Windows conventions To simplify operation you can use tool buttons icons in the measurement menu in this case you do not need to open a pulldown menu The tool buttons and their functions Function Save measured data Clear sample matrix for new measurement Copy measured data Show measured data in Excel Enter comment on measure ment Button for selection of wells Print Start measurement Stop measurement Move microplate to home posi tion Run background measurement Pulldown menu gt Option File gt Save as File gt New Edit gt Copy Edit gt Send to Excel Edit gt Measurement Comments Measure gt Select wells to Measure File gt Print Measure gt Start Measurement Measure gt Stop Measurement Measure gt Move home Measure gt Get Background Microplate Luminometer Structure of the Software EE Tabulated presentation of r
87. tervals that are long enough to allow for the microplate transport 65 Microplate Luminometer 66 Slow Kinetics The program automatically corrects for intervals that are too short If you have defined an interval that is too short the program calculates the required intervals using the new time actually required and displays it on the screen after measurement of the 1 data point The measurement can then be continued with the new parameters or aborted For example if you start a measurement with 2 samples using the parameters from Fig ure 6 2 the time data is changed upon completion of the 1 in terval depending on the position of the selected wells the data points are e g 0 s 6 s 12 s instead of Os 1 s 2 s Well Timing tab with injectors Except for the measurement time enter the parameters for the individual injections here When using one injector Injector 1 Injector 2 uf Microplate yp Or Select the injector inj 1 or inj 2 by clicking on the respective box Then enter the injector parameters Please note Injector 1 injects into the well before the meas urement position injector 2 in the measurement position Volume Enter the injection volume in ul via keyboard or by clicking on the arrow buttons Injection 1 to measurement delay Enter the delay time be tween the injection of injector 1 and the start of the measure ment in seconds The hardware dependent mini
88. the last measurement have not yet been saved In all these cases no measurement can be started If the instrument door is not closed a message is displayed in dicating the current measurement mode Le pF Door open error ERR_OUOR_UPEN Close the instrument door and click lt Repeat gt or lt OK gt Then the measurement will be started Any programmed injections will be performed and the selected wells are measured one after the other The results of the indi vidual samples are entered into the respective fields 61 Microplate Luminometer 62 Measurement end Measurement stop Raw Data The measurement is finished as soon as all selected wells have been measured In the autosave mode the measured re sults are automatically saved and the measured values cleared from the screen If the autosave mode is not active the meas ured values remain displayed and can be saved Push the Stop button red signal light to stop a measurement The results measured so far can be stored Microplate Luminometer Slow Kinetics 6 Slow Kinetics 6 1 Overview In the measurement mode Slow Kinetics the trend of the light emission of samples is measured To this end the number of data points must be defined The measurement starts after in put of the measurement parameters and definition of the wells to be measured Then all samples for data point 1 are meas ured in succession and the averaged results displayed in a ta
89. tled Raw Data PC a ie EE Fille Edit Options Measure View Help Hie ace ala ole eee Background 0 Date lon 729 99 Injection T volume 10pl Injection 2 volume 10l Eo ine a Een dees xl apt 2 J d e ts je a RN SO COO COO a a a a a a a ee ee ee i Tou must initialize the injectors before you can measure Please note Before running a measurement if desired select the autosave mode Select this function in the Options menu Procedure without injections Ll Insert microplate correctly into the luminometer and close cover Li Run background measurement if necessary Zech L Select the wells to be measured mh LI Start measurement Microplate Luminometer Measurement start Measurement procedure Raw Data Procedure with injections LI Select the wells to be measured LI Place plastic tub in plate loading compartment O Initialize injectors 24 LJ Prime injector tubing if necessary it Insert microplate LI Insert microplate correctly in luminometer and close door LI If necessary run background measurement GB LI Select the wells to be measured E O Start measurement Measurement of the microplate or the selected area starts after pressing the Start button green signal light when the instru ment door is closed The Start button is dimmed gray when a measurement is run ning or no communication has been established with the lumi nometer or the data of
90. turns to the Protocol Manager and shows the name of the new measurement protocols in the field Runable protocols Microplate Luminometer lt Save gt lt Cancel gt Fast Kinetics Saves the parameters in the existing measurement protocol file After you have saved the protocol the program returns to the Protocol Manager The changes are disregarded and the old values are still valid The program returns to the Protocol Manager 83 Microplate Luminometer 84 Figure 7 7 Measurement menu Fast Kinetics wit sample matrix and table of measured values Fast Kinetics 7 3 Measurement As soon as you have selected the measurement protocol type Fast Kinetics and you have clicked lt Run gt the program changes to the measurement menu Fast Kinetics Communi cation with the Microplate Luminometer is established provided this has not already been done at the start of the program Ini tializing instrument appears on the status bar and the yellow LED lights up In the measurement menu Fast Kinetics the sample matrix is displayed first and below that a table for the measured values The table includes row by row the predefined data points for all samples on the microplate see Figure 7 7 Ameasurement of the entire plate can also be started immediately by pressing the Start button Fast Kinetics PC Untitled Data Table S File Edit Options View Measure window Help lej x a a Elm EEN A aA
91. urn instrument on LI Check if green LED is on If the green LED is not on please contact our Service Depart ment Microplate Luminometer Figure 8 6 Power supply unit on the rear panel of the injector unit Maintenance 8 14 Changing the Operating System Voltage in the Injector Unit You need 1 small flathead screwdriver Spare fuses For operating voltage 230 V Fuse 160 mA slow blow For operating voltage 115 V Fuse 315 mA slow blow Use only UL approved fuses ee a a k amp N i eeCoscas ee o G sS lt a Fuse holder Recess for screwdriver LJ Turn main switch to off position 0 LJ Disconnect power cord from wall outlet LI Insert a screwdriver or a similar tool into the recess on the right side of the fuse holder push slightly and take the fuse holder out The safety holder includes one fuse each for op erating voltage 115 V and 230 V LJ Turn the fuse holder by 180 and insert the fuse holder again such that the label indicating the respective operating voltage is visible in the window 109 Microplate Luminometer Technical Data 9 Technical Data MONOLIGHT 3096 Microplate Luminometer Sample format 96 well microplate non transparent fixed or strip format Detector Photomultiplier with bialkali cathode 300 650 nm Sensitivity Less than 50 attomol ATP depending on reagents Crosstalk Less than 3 x 10 with 96 well microplates Dynamic
92. were manu factured in accordance with the safety requirements for elec tronic and medical measuring systems If the law states regula tions on the installation and or operation of sample measuring systems then it is the operator s responsibility to adhere to them The manufacturer has done everything possible to guarantee that the equipment functions safely both electrically and me chanically The user must ensure that the instruments are set up and installed in such a way that their function is not impaired The instruments are tested by the manufacturer and supplied in a condition that allows safe and reliable operation This Users Manual includes information and warnings that must be observed by the user in order to ensure safe operation of the instruments Please adhere to the following safety instructions when han dling or operating the system 1 The instruments may only be operated by personnel who have been trained on the use of the systems It is strongly recommended that all users read this manual prior to use 2 Use the instruments only for the designated application 3 BD Biosciences Pharmingen assumes no liability for any damages including those to third parties caused by improper use or handling of the instruments 4 The user may only perform the maintenance work described in this manual 5 Use only the parts described in this manual for servicing Microplate Luminometer CAUTION Safety Instruct

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