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3

Operating Procedures

This chapter explains how to start up the system and how to use the
control panel and the SoftMax Pro Software to configure instrument
settings, define experiment protocols, and run an analysis, as described
in the following sections.

•

Overview, see page 66

•

Starting Up the System, see page 67

•

Setting the Temperature, see page 72

•

Setting Up the Software, see page 75

•

SoftMax Pro Software Parameters, see page 78

•

SoftMax Pro Software Parameters for Fluid Transfer, see page 86

•

Other Software Settings, see page 99

•

Reading a Microplate, see page 99

•

Shutting Down the System, see page 103

•

Optimizing Fluorescence Assays, see page 103

•

Optimizing Time-Resolved Fluorescence Assays, see page 110

•

Optimizing Fluorescence Polarization Assays, see page 111

•

Optimizing Luminescence Assays, see page 112

Note:

The information in this chapter assumes that the instrument and

computer are properly installed and connected. See

Installation on

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Summary of Contents for FlexStation 3

Page 1: ...www moleculardevices com FlexStation 3 Benchtop Multi Mode Microplate Reader User Guide 0112 0127 B June 2013 FlexStation_manual book Page 1 Tuesday June 25 2013 10 22 AM...

Page 2: ...or their product names as trademarks Each product is shipped with documentation stating specifications and other technical information Molecular Devices products are warranted to meet the stated spec...

Page 3: ...ers and Instrument Panels 17 Drawers 21 Fluidics Module 24 Detection Module 26 Computer 28 Accessories 28 Consumables 30 Overview of Operation 32 Choosing an Experiment 32 Preparing the Instrument 32...

Page 4: ...nd Baseplate Installation 63 Installing SoftMax Pro 63 Chapter 3 Operating Procedures 65 Overview 66 Starting Up the System 67 Using the Control Panel 70 Setting the Temperature 72 Displaying the Temp...

Page 5: ...Wavelengths for Fluorescence Assays 105 Optimizing Time Resolved Fluorescence Assays 110 Optimizing Fluorescence Polarization Assays 111 Optimizing Luminescence Assays 112 Chapter 4 Maintenance 113 O...

Page 6: ...Opening the Instrument 137 Evaluating the Tip Rack 138 Inspecting Inside the Fluidics Module 139 Removing the Pipettor Head 142 Expelling Undispensed Fluid from Tips 145 Recovery Procedure 146 General...

Page 7: ...of increasing the liquid handling throughput and flexibility for biochemical based and cell based assays Using an integrated 8 channel or 16 channel pipettor increases assay flexibility by transferri...

Page 8: ...system to provide flexibility to detect multiple detection modes while a separate PMT provides additional sensitivity for luminescence applications Reference diodes automatically adjust to slight fluc...

Page 9: ...nce on page 113 provides instructions for cleaning the fan filter changing the fuses and moving the system to a new location Troubleshooting Procedures on page 129 provides instructions for diagnosing...

Page 10: ...caution is an instruction that if not followed can result in damage to the system or in loss of data WARNING A warning is an instruction that if not followed can result in potential injury to a person...

Page 11: ...l nature be sure to read all sections of this user guide before attempting a reading with fluidics See Understanding Potential Mechanical Problems on page 133 Safety Messages Observe the following war...

Page 12: ...ure the system is connected to a power receptacle that provides voltage and current within the specified rating for the system Use of an incompatible power receptacle can create electrical shock and f...

Page 13: ...conjunction rather than individually Integrated pipetting provides flexibility and also offers users parameters to optimize assay robustness Trituration mixing via aspiration and dispense of the pipet...

Page 14: ...ltraviolet spectra must be used A plate drawer adapter is provided with the reader The adapter is required for optimum performance when reading from the top or bottom of standard 96 well and 384 well...

Page 15: ...more The extreme flexibility and high sensitivity of the FlexStation 3 reader makes it appropriate for applications within the fields of biochemistry cell biology immunology toxicology molecular biol...

Page 16: ...exStation 3 Benchtop Multi Mode Microplate Reader User Guide 16 0112 0127B Figure 1 3 Instrument Front View Figure 1 4 Instrument Rear View FlexStation_manual book Page 16 Tuesday June 25 2013 10 22 A...

Page 17: ...tions of the detection module Figure 1 5 Lifting Off the Top Cover Note The top cover can be lifted back as shown in the figure below for certain limited troubleshooting procedures See Opening the Ins...

Page 18: ...initiate and regulate the temperature and to open and close the drawers A 2 3 character liquid crystal display LCD shows the current instrument temperature at all times and the set point temperature...

Page 19: ...box cover and power cord receptacle The lower panel consists of an RS 232 serial port and parallel port not currently active There are also a number of identification labels Figure 1 7 Input Output P...

Page 20: ...cover provides access to the flash lamp on the right side of the instrument as viewed from the rear Figure 1 8 Rear View See Replacing the Flash Lamp on page 123 CAUTION Flash lamp access and maintena...

Page 21: ...t of the instrument The reading chamber drawer in the detection module transports the assay microplate into the reading chamber Figure 1 9 Instrument with Drawers Open and Carriages Accessible Small p...

Page 22: ...drawer or Open Drawer for the reading chamber drawer Figure 1 11 SoftMax Pro Software Control Menu You can also open or close the reading chamber drawer with the Drawer button on the Status Bar Figur...

Page 23: ...is in the reading chamber It is the lowest of the three drawers The reading chamber drawer operation varies depending on the incubator status When the incubator is off the reading chamber drawer is op...

Page 24: ...d moves vertically between the drawers The fluidics module can be opened by service trained users from the inside front panel if necessary for maintenance or from the inside top panel to install or re...

Page 25: ...head for use with 384 well microplates Figure 1 14 Pipettor Head See Installing the Pipettor Head on page 56 The barrels on the pipettor head require periodic cleaning to remove silicone lubricant dus...

Page 26: ...ntenance The detection module is contained in a molded plastic housing to which the top cover is attached at the back of the instrument Figure 1 15 Detection Module Detail Reading Chamber The reading...

Page 27: ...ading chamber They are the excitation fibers thin and black or red has a collar and pins emission fibers black and fatter with attached electrical cord electrical connector to the read head green with...

Page 28: ...omputer configuration includes a Pentium processor with 2 8 GHz 1 GB hard drive See Installing SoftMax Pro on page 63 and Setting Up the Software on page 75 Accessories The following accessories are i...

Page 29: ...etween these computers and the instrument using a USB to serial adapter Molecular Devices has tested many third party USB to serial adapter cables and has found Keyspan USA 19HS Molecular Devices P N...

Page 30: ...he microplate can vary depending on the type of microplate used Microplates supported for use in this reader are 6 well 12 well 24 well 48 well 96 well and 384 well standard formats 96 well half area...

Page 31: ...e microplates can optimize light collection Note Not all microplates are made with the same materials Some plastics most notably polystyrene have significant native fluorescence and can cause moderate...

Page 32: ...ng partial cases Tips are available in both black and clear options Black tips are generally used during fluorescence assays when auto fluorescent properties of clear tips can interfere with your resp...

Page 33: ...Assay Read Types see page 38 Instrument Design Fluidics The instrument is designed with a fluidics module that transfers liquids to the assay plate during a fast kinetic read or before an experiment...

Page 34: ...gent volumes for fast kinetic cell based assays such as calcium mobilization The Automix function permits automatic shaking of the microplate at preset intervals thereby mixing the contents within eac...

Page 35: ...ient Temperature regulation and control of the reading chamber is achieved through electric heaters a fan efficient insulation and temperature sensors The heaters are located within the instrument whi...

Page 36: ...important factor in fast kinetic assays For this reason all readings are tagged with an exact read time and when multiple well fast kinetic responses are plotted the curves overlie each other as plot...

Page 37: ...luorescence values For more detail of the optical design and an illustration of the optical system see Detection Module on page 26 Bottom and Top Reading Switching to bottom or top reading capability...

Page 38: ...selected total experimental time At a preselected point or points during that time sequence the pipettor can transfer up to three reagents from the compound plate to the assay plate The instrument con...

Page 39: ...ce Fluorescence Luminescence and Time Resolved Fluorescence Absorbance Fluorescence and Luminescence System Settings Top read Yes or No Yes or No Yes or No Yes or No No Bottom read Yes or No Yes or No...

Page 40: ...up parameters selected in the SoftMax Pro Software but is limited to 2 hours and 45 minutes 165 minutes At the end of a reading rates are reported as each well Kinetic analysis has many advantages whe...

Page 41: ...roplate wells this non confluent growth pattern can require multiple reads at different locations in a well When used with 6 well 12 well 24 well 48 well or 96 well plates well scanning allows maximum...

Page 42: ...omix before or between reads Automix shakes the microplate at preset intervals thereby mixing the contents of each well highly recommended for ELISAs and other solid phase enzyme mediated reactions No...

Page 43: ...and read plates Acquire Data from the Reader Pre read microplates Analyze kinetic and spectrum data as it is collected Save data files for in depth analysis at a later time Save multiple microplates w...

Page 44: ...nst a standard curve Analyze and compare data within a plate between plates and between experiments Customize your print formats Print all or individual sections of the data file Define and print a re...

Page 45: ...during relevant steps You can control the instrument by using either buttons and icons in the windows and along the tool bars or by using the menus You can use either your mouse or keystrokes to make...

Page 46: ...ve different icons Table 1 2 SoftMax Pro Software Status Bar Icons Icon Description The Instrument Status icon provides visual confirmation that SoftMax Pro is communicating with the instrument Double...

Page 47: ...le 1 3 Icon Description Double click the Plate icon to open the Plate section in a new window Double click the Name of Plate icon to open the Section dialog Click the Settings button to open the Instr...

Page 48: ...FlexStation 3 Benchtop Multi Mode Microplate Reader User Guide 48 0112 0127B FlexStation_manual book Page 48 Tuesday June 25 2013 10 22 AM...

Page 49: ...les see page 61 Installing the Drawer Adapters see page 62 Installing SoftMax Pro see page 63 General Precautionary Information CAUTION The following procedures must be completed by a service trained...

Page 50: ...it it is particularly important that you retain them for inspection by the carrier in case there has also been damage to the instrument As you unpack the system components examine the packing list tha...

Page 51: ...ch the midsection of the crate on top and bottom and then move that midsection aside Due to the size of the crate this step requires two people 2 Slide the plastic bag enclosing the instrument out of...

Page 52: ...hold down the cover of the instrument Figure 2 1 Location of Shipping Screws 2 Use the 3 32 inch hex key to unscrew the shipping screws It might be necessary to move the instrument to the front of the...

Page 53: ...re also two quarter turn fasteners Zeus screws attached to the flange and two locating pins near the middle of the reading chamber Figure 2 2 Positioning the Fluidics Module CAUTION To avoid damage to...

Page 54: ...he bottom of the fluidics module 3 Connect the quarter turn fasteners to the bottom of the fluidics module and lock the fasteners into place 4 Attach the restraining cable from fluidics module to the...

Page 55: ...that the instrument can be serviced without first detaching the communication or power cable 5 With the fluidics module tilted back connect the 15 pin sub D electrical connector into the communication...

Page 56: ...r Head After you install the fluidics module you can place the pipettor head into the fluidics module Use this same procedure for both the 8 channel and 16 channel pipettor heads The barrels on the pi...

Page 57: ...0127 B 57 2 Turn the quarter turn fastener on the inside top cover and then unfold the cover off the fluidics module Figure 2 7 Opening the Inside Top Cover FlexStation_manual book Page 57 Tuesday Jun...

Page 58: ...pipettor head into its approximate position under the z stage plate and red mounting knob 5 Maneuver the cable down toward the back of the cavity and align the connector over the receptacle 6 Press th...

Page 59: ...sitioning Pipettor Head 8 With one hand pull up on the red knob 9 With the other hand align the metal plate at the back of the pipettor head with the screw hole and the two locating pins underneath th...

Page 60: ...Head Installed 13 Fold the inside top panel back over the pipettor head and lock the quarter turn screw in place 14 Bring the top cover back over the fluidics module and snap it into place at the hand...

Page 61: ...t at the wall Connecting the Cables After the instrument is assembled and the computer is set up you can connect computer cable and the power cord Figure 2 11 Computer Cord and Power Cord Locations 1...

Page 62: ...microplate adapter and metal compound baseplate Microplate Adapter Installation To bottom read or top read a standard 96 well or 384 well microplate you must install the black microplate adapter in t...

Page 63: ...corner of the drawer Installing SoftMax Pro Install the SoftMax Pro Software on the computer according to the instructions in the SoftMax Pro Software User Guide CAUTION Always remove any microplates...

Page 64: ...FlexStation 3 Benchtop Multi Mode Microplate Reader User Guide 64 0112 0127B FlexStation_manual book Page 64 Tuesday June 25 2013 10 22 AM...

Page 65: ...Software Parameters see page 78 SoftMax Pro Software Parameters for Fluid Transfer see page 86 Other Software Settings see page 99 Reading a Microplate see page 99 Shutting Down the System see page 10...

Page 66: ...ther instrument parameters See Setting the Temperature on page 72 3 Use the SoftMax Pro Software to configure the read mode type of analysis template and so on as desired You can also create sections...

Page 67: ...ss the rocker switch to the ON position I The instrument automatically performs diagnostic checks to ensure that it is functioning correctly All three drawers open and close After about four minutes t...

Page 68: ...Double click the SoftMax Pro Software icon to start the program The SoftMax Pro Software opens with an Untitled window in Flex mode with a default template selected Note No set point temperature appea...

Page 69: ...e tutorial described in this window Also refer to the SoftMax Pro Software User Guide For users familiar with SoftMax Pro Software If you are already familiar with SoftMax Pro Software you can close t...

Page 70: ...3 4 Detail of Control Panel with LCD The following tables describe the indicators and buttons in the control panel Table 3 1 Indicators in the Control Panel Indicator Description The actual C provides...

Page 71: ...e to increase or decrease the temperature shown in the display by an increment of 0 1 C press and hold to scroll the temperature The tip rack button opens or closes the tip rack drawer The reagents bu...

Page 72: ...erature cannot be regulated at a set point that is outside this range Displaying the Temperature Two temperatures are displayed in the LCD on the control panel Figure 3 5 Temperature Control and Displ...

Page 73: ...ess the up or down arrow keys until the desired temperature set point appears on the display The reading chamber temperature is maintained at the set point until you disable temperature control by tou...

Page 74: ...r Button The Incubator dialog appears Figure 3 8 Incubator Dialog You can leave the temperature setting at the default value or you can type a different value into the Temperature field Note The incub...

Page 75: ...ar If there is a red X in front of the Instrument Status icon if there is no temperature in the Temperature display field or if you see other problems you might need to adjust instrument preferences T...

Page 76: ...ation between the computer and the instrument or to resolve a different instrument problem See Troubleshooting Procedures on page 129 6 The SoftMax Pro application defaults to Flex mode every time you...

Page 77: ...plate section with settings identical to a particular existing plate section select that plate section and then click Edit Duplicate Double click on the word Plate in the Plate Section tool bar to op...

Page 78: ...er Guide and they are discussed in more detail in the SoftMax Pro Software User Guide All possible combinations of type mode and parameter are not presented here but Molecular Devices technical suppor...

Page 79: ...me sample at many different wavelengths within the range specified No fluid integrated transfer is available Well Scan Multiple readings are taken in different areas of each well in a 6 well to 96 wel...

Page 80: ...er of readings results in better reader precision In fluorescence intensity reading the sample can photobleach if a high number of flashes are used so if the plate will be re read especially for kinet...

Page 81: ...e the default values click in the appropriate box and then type the desired run time or interval The acceptable run time range is from 0 seconds to 1000 seconds After you have entered the values the t...

Page 82: ...after you have selected settings that follow for example Compound Source Compound and Tip Columns the earlier settings might be automatically reconfigured to reflect the new assay plate type and well...

Page 83: ...Select the wells you want read by dragging the pointer Wells must be contiguous and in a rectangular arrangement but do not need to start in the column 1 You can choose a partial row or column or a si...

Page 84: ...x function is a patented feature that allows you to set automated shaking of the microplate before and between readings Select Before First Read to shake the microplate before the first wavelength rea...

Page 85: ...has been moved to begin the read of another column This delay allows time for the meniscus motion to cease potentially improving precision especially in the low density high volume microplates with f...

Page 86: ...Type Wells to Read Compound Source Compound Transfer and Compound Tip Columns must correlate The SoftMax Pro Software cross checks these settings as you move through the configuration windows and aft...

Page 87: ...number of wells selected in the Assay Plate Type setting In particular the well bottom height is different for different types of plates and selecting the plate type correctly is important to prevent...

Page 88: ...periments this setting also helps prevent flooding of the assay plate The fields in this dialog allow you to set volumes for up to three transfers However you must be careful to keep in mind the actua...

Page 89: ...e compound transfers do not type a value in this field Any value entered is saved with the file but the value is not displayed anywhere except in the Compound Transfer setting Values for a 384 well mi...

Page 90: ...nt number increases in increments of 26 microliters Therefore a setting of 2 is equal to 52 L per second A setting of 3 or 4 can help minimize cell damage or dislodgment In 384 well microplates the sp...

Page 91: ...t event and adds the total time necessary to load pipette tips aspirate new fluids from the compound plate and dispense them into the assay plate Possible Problems Time point calculations are based on...

Page 92: ...r Compound Source or Assay Plate or both You can set the Volume of fluid to be withdrawn from the well and the number of times Cycles to be aspirated and dispensed into that well In addition for assay...

Page 93: ...n one or more transfers are enabled these settings allow you to choose the tips and compounds to be used for transfers The settings are either automatically assigned by the software or you can manuall...

Page 94: ...ed from left to right The read transfer read sequence in each column is initiated only after the previous column s read transfer read event is completed the total read time for that column The fluid t...

Page 95: ...hrough 12 This settings show one two or three transfers with blue pink and green color coded tips to match the setting made in the Compound Transfer window There are two menus Tips Column and Compound...

Page 96: ...vent Using the color associated with each event the compound settings display the dispensed compound volume as a percentage of the total assay well volume If you entered an initial volume in the Compo...

Page 97: ...he liquid in the tip Figure 3 29 Pipette Tip Air Gap To set the pipette tip air gap click Control Set Air Gap Figure 3 30 Setting Pipette Tip Air Gap In the Air Gap Settings dialog the allowed values...

Page 98: ...For example the following figure shows a Plate section with three fluid transfers The blue squares in their upper left corners represent the first transfer the red squares in the upper right corners r...

Page 99: ...ormation about using the software to continue to prepare for an experiment see the SoftMax Pro Software User Guide Reading a Microplate Loading Tips and Microplates Prepare your tip rack compound plat...

Page 100: ...ll microplates For more information see Using the Microplate Adapters on page 120 and Using the Compound Baseplate on page 121 3 Close drawer by either pressing the appropriate drawer button on the in...

Page 101: ...active starting a read will read the active plate Replacing Data in a Plate If you select a Plate section that already contains data and select the Read command an alert appears requiring you to confi...

Page 102: ...ay of the Plate section as they are received from the instrument to the software in real time For information about how to change the data display see the SoftMax Pro User Guide Table 3 3 Status Bar R...

Page 103: ...elengths emission cutoff filter readings per well the PMT voltage and the temperature of the reading chamber Another important factor that is independent of the instrument but which affect assays opti...

Page 104: ...ecise measurements The default number of readings per well varies with the read mode For fluorescence the default is 6 and for luminescence the default is 30 PMT Voltage The voltage of the photomultip...

Page 105: ...value obtained from the literature or the customary excitation filter 4 Set the step increment to 2 nm or 3 nm You can choose to do a preliminary scan with a 10 nm increment to determine the approxim...

Page 106: ...mission scan to start stop approximately 50 nm below or above the tentative emission value obtained from the literature or existing filter pair 4 Set the step increment to 2 nm to 3 nm You can choose...

Page 107: ...tting the wavelength to that determined in Step 6 2 Perform the scan and view the results as a plot of fluorescence vs emission wavelength Note the wavelength giving the maximum emission the optimal e...

Page 108: ...rs manually For emission wavelengths less than 325 nm experimental iteration is usually the best method of determining the optimal emission and excitation wavelengths Begin optimization by performing...

Page 109: ...ion cutoff filters For Spectrum reads the default is manual no automatic cutoff Table 3 4 Emission Cutoff Filter Default Settings Number Automatic Cutoff Selection Wavelength nm Endpoint and Kinetic R...

Page 110: ...and speed are achieved Important settings for obtaining the best results in TRF assays are integration delay and integration time The integration delay is the amount of time that elapses between the f...

Page 111: ...f the signal in the background wells is more than 1 10 the signal in the wells containing fluorophore then background wells should be run on each assay plate The average raw signal from the parallel a...

Page 112: ...best sensitivity should be achieved using the standard luminescence measurement without a wavelength selected Luminescence read times are not designated by multiple reads per well but rather by choosi...

Page 113: ...ety and to prevent instrument damage Example WARNING All maintenance procedures described in this user guide can be safely performed by qualified personnel Maintenance not covered in this user guide s...

Page 114: ...18 944 8000 Please have the system ID number system serial number software version number and the system owner s name available when you call Moving the Instrument If you need to relocate the FlexStat...

Page 115: ...he FlexStation 3 instrument using a cloth or sponge that has been dampened with water Do not use abrasive cleaners If required clean the surfaces using a mild soap solution diluted with water or a gla...

Page 116: ...reader OFF and then remove the power cord and cables from the back of the reader 3 Turn the reader over so that it rests flat on the bench 4 Remove the black fan cover and then remove the filter 5 Cle...

Page 117: ...and OS 2 silicone cleaner or alcohol Do not touch the barrels with your hands during cleaning To clean the barrels on the pipettor head 1 Turn off the power to the instrument and unplug the power cord...

Page 118: ...etainer nut Figure 4 3 Removing the Pipettor Head 5 Slide the pipettor head part way out from under the metal mounting plate and of the top panel opening supporting it from underneath 6 Lift the pipet...

Page 119: ...making sure that the white spiral cord is secured on its retaining hook Figure 4 4 Pipettor Head Coil Hook 10 Tighten the red retainer nut to secure the pipettor head Make sure the knob is as tight as...

Page 120: ...ware 3 Lower the black adapter plate into the reading chamber drawer with its cutout corner facing the front left corner of the drawer Removing the Microplate Adapter If you are reading high profile 6...

Page 121: ...loose or disconnected and that power to the power outlet is functioning properly If these checks fail to remedy the loss of power follow the steps listed to replace the fuses Spare fuses two U S and...

Page 122: ...see that it contains two fuses It is possible that only one of the fuses has blown Molecular Devices recommends that you replace both fuses however to ensure continued proper operation 7 Pull both fu...

Page 123: ...ver Figure 4 7 Flash Lamp Location WARNING The following procedures must be completed by a service trained user Do not attempt the following procedures if you have not been trained properly by appropr...

Page 124: ...e power cord and the computer cable from the rear of the instrument 2 Remove the Phillips head screw on the top of the cover and lift the cover off vertically 3 Set the cover aside and keep track of t...

Page 125: ...ated toward the inside of the lamp s mounting box The two cap screws are spring loaded and stay in a raised position so that you can slide out the entire Litepac without getting the screws caught in t...

Page 126: ...move the two screws that hold down the diamond shaped retainer Be careful not to lose the screws Figure 4 11 Withdraw the Lamp 9 Withdraw the silver colored lamp from its multi pin socket by gently ro...

Page 127: ...p with the bore of the mounting block 17 Slide the lamp all the way into the block orienting the assembly to clear the baseplate Slide it in until it stops 18 Tuck the orange power cord inside behind...

Page 128: ...strument for an extended period of time clean the external surfaces of the instrument before storage See Cleaning the Instrument on page 115 Be sure the instrument is emptied of all plates and tips be...

Page 129: ...when Using Fluidics see page 135 General Error Messages see page 147 Tilting or Removing the Fluidics Module see page 149 Note These troubleshooting instructions address only those problems that can...

Page 130: ...n on the right there is either a communication problem between the instrument and the software or the instrument has experienced a fatal error 1 Make sure that the FlexStation 3 instrument is plugged...

Page 131: ...The communication speed is 9600 during the instrument s initial warm up sequence while it is first communicating with the software If you continue to see the red X after confirming the Serial Comm Spe...

Page 132: ...ter cables from the computer 2 Using your fingernail or a narrow blade open the drawer door to its horizontal position and pull out the carriage slowly and as gently as possible WARNING The power must...

Page 133: ...rels See Cleaning the Barrels on the Pipettor Head on page 117 If the power fails during a pipetting step serious damage can result to the instrument if appropriate corrective steps are not taken As n...

Page 134: ...Remove the lid of the tip rack before placing it in the tip rack drawer Make sure that you seat the tip rack plates and trough if applicable securely in the correct drawers Make sure that the layout...

Page 135: ...he computer screen indicating a problem with the instrument You might need to open the instrument to locate and correct the problem The procedures in this section include Assessing a Mechanical Proble...

Page 136: ...of the problem Fluidics module not detected This message might occur if the fluidics module is not correctly installed Pipettor failure There may be undispensed fluid in the pipette tips This message...

Page 137: ...he four quarter turn screws and lift the front panel out You might need to inspect the interior of the module to determine if the pipettor was in the process of a fluid transfer when the power was int...

Page 138: ...ght be able to remove the rack and problematic tips from outside the fluidics module by pulling the tip rack all the way out If not gently push the tip rack back inside the instrument Reach in from th...

Page 139: ...e compounds plate area or the reading chamber Are there any pipette tips on the pipettor head Is there is any chance there is liquid remaining in the tips If so remove the head and expel the liquid sa...

Page 140: ...the rack out Pipettor head is in home position without tips but tips are missing If the pipettor head is up and out of the way without tips but there are tips missing from the rack then you must locat...

Page 141: ...ight After you have secured all tips and returned the tip rack and compound plate drawers to their outside right most position replace the inside front panel and top cover Reattach the power cord and...

Page 142: ...s turned off 2 Unplug the power cord and the computer cable from the rear of the instrument 3 Open the instrument top cover Figure 5 4 Opening the Instrument Cover 4 Turn the quarter turn fastener on...

Page 143: ...Location of the Pipettor Head Inside the Fluidics Module 6 Slide the pipettor head part way out from under the metal mounting plate and of the top panel opening supporting it from underneath FlexStati...

Page 144: ...o dispense liquid from the tips skip the next two steps and see next Expelling Undispensed Fluid from Tips on page 145 8 To completely remove the pipettor head unscrew the black connector at the end o...

Page 145: ...uidics module making sure to secure the white spiral cord back on its retaining hook 2 Tighten the red retainer nut to secure the pipettor head Make sure the knob is as tight as possible 3 Replace the...

Page 146: ...ro Status bar then see Problems During Startup on page 130 If the temperature display on the front panel of the instrument displays the actual current temperature but the red X is still covering the i...

Page 147: ...se messages are generally self explanatory For assistance regarding warning messages contact your local Molecular Devices representative Table 5 2 FlexStation 3 Instrument Error Code Ranges Error Code...

Page 148: ...ng messages attempt your reading again before continuing with troubleshooting Unexpected response Result Code 105 Instrument communication problem Response too late Measurement aborted Result code Ins...

Page 149: ...ing procedures if you have not been trained properly by appropriate Molecular Devices personnel WARNING Turn off the power switch and disconnect the power cord from the main power source before perfor...

Page 150: ...gure 5 8 Tilting the Fluidics Module If you can clean up the spill or remove the tips without removing the fluidics module do so now If you need to remove the fluidics module completely see Removing t...

Page 151: ...9 Tilting the Fluidics Module 4 Locate the electrical connector between the bottom of the fluidics module and the housing and then unscrew the retaining screws and unplug the connector 5 Locate the fl...

Page 152: ...might be able to see tips inside If you see tips all the way down in the bottom tray of the instrument you can safely leave them there They will be out of the way of the operation of the read heads 9...

Page 153: ...uter cable 0310 9336 Bottom read microplate adapter reading chamber 2500 1029 Compound Baseplate compound plate drawer 4600 0029 Fuse 0310 3992 Pipettor head 8 channel 0310 5025 Pipettor head 16 chann...

Page 154: ...sories contact Molecular Devices Customer Service or your authorized service representative Molecular Devices 1311 Orleans Drive Sunnyvale California 94089 Telephone 1 408 747 1700 Fax 1 408 747 3601...

Page 155: ...volume in the microplate well is between 100 L and 300 L Technical specifications are subject to change without notice Table B 1 Fluidics Specifications for the 8 Channel Pipettor Item Specification V...

Page 156: ...ric resolution 0 001 OD Photometric accuracy linearity 0 OD to 2 0 OD 0 006 OD 1 0 Photometric precision repeatability 0 OD to 2 0 OD 0 003 OD 1 0 Stray light 0 05 at 230 nm Photometric stabilization...

Page 157: ...well black plates auto gain circuitry System validation Self calibrating with built in fluorescence calibrators Light source Xenon flash lamp 1 joule flash Average lamp lifetime 2 years normal operat...

Page 158: ...oplates top read Wavelength range 250 nm to 850 nm Crosstalk 0 3 in 96 well and 384 well microplates Table B 8 Photometric Analysis Mode Using SoftMax Pro Software Item Specification Data expression A...

Page 159: ...fter initiation of temperature regulation Temperature regulation 4 sensors Temperature regulation diagnostics Temperature regulation system is continuously monitored and updated Evaporation Plate lid...

Page 160: ...s on board diagnostics Spill control Drawer mechanism and reading chamber assembly protected from accidental spillage by drainage ports Computer interface 8 pin DIN RS 232 serial double shielding requ...

Page 161: ...rating humidity 0 to 70 non condensing Storage temperature 20 C to 65 C Table B 16 Physical Specifications Item Specification Size H x W x D 48 cm 58 cm 41 cm 19 in 23 in 16 in Weight 22 7 kg 50 lbs P...

Page 162: ...FlexStation 3 Benchtop Multi Mode Microplate Reader User Guide 162 0112 0127B FlexStation_manual book Page 162 Tuesday June 25 2013 10 22 AM...

Page 163: ...he PMT In automatic mode the instrument sets the cutoffs automatically based on the wavelengths chosen for reading In manual mode you can choose the filter wavelength Emission Spectrum Scan An emissio...

Page 164: ...ectrum of excitation wavelengths The default value reported for each well is the wavelength of maximum fluorescence excitation in RFU Fluorophore A fluorophore is a material that absorbs light energy...

Page 165: ...T A Photomultiplier Tube PMT is a vacuum tube that detects light from dim sources through the use of photoemission and successive instances of secondary emission to produce enough electrons to generat...

Page 166: ...lifetime Labels with prolonged fluorescence lifetime The time gating electronics introduce a delay between the cut off of each light pulse and the start of signal accumulation During the delay the uns...

Page 167: ...omix 46 Compounds 63 121 Drawer 46 FLEX 77 Incubator 46 74 Read 46 100 102 Reading Chamber 62 120 Setup 47 76 Stop 46 102 Transfer 89 C cables 29 117 121 142 153 cleaning the instrument 115 components...

Page 168: ...lamp 20 124 127 removing shipping screws 52 tip rack 138 top 17 52 57 60 117 125 127 137 142 150 D data appearing in Plate section 102 display parameters 99 replacing with new 101 detection module 24...

Page 169: ...utoCalibrate selection 85 Automix 84 AutoRead 85 compound and tip columns 93 compound source 87 compound transfer 88 timing 81 triturate selection 92 wells to read 83 Instrument Status icon 46 75 L la...

Page 170: ...Plate Section screen 84 98 Plate sections 102 creating and editing 77 selecting 101 power cord 19 28 117 142 153 connecting 61 power failure 135 power switch 19 67 Preferences dialog 75 printer cable...

Page 171: ...88 on Plate Section screen 98 pipette tip air gap 97 timing 81 triturate selection 92 wells to read 83 Setup button 47 76 shipping containers 50 shutdown procedures 103 long term 128 SoftMax Pro 28 in...

Page 172: ...inspecting inside 139 fluidics module tilting or removing 149 general error messages 148 mechanical problems Flex mode 135 opening drawers manually 132 opening the instrument 137 pipettor head removin...

Page 173: ...FlexStation_manual book Page 173 Tuesday June 25 2013 10 22 AM...

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