Glomax Multi Plus Detection System Protocol

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Promega Corporation 2800 Woods Hollow Road Madison, WI 53711-5399 USAToll Free in USA 800-356-9526 Phone 608-274-4330 Fax 608-277-2516 www.promega.comPrinted in USA. Part# TM311Revised 5/10 Page 1

1. Description..........................................................................................................3A. Inspection...............................................................................................................4B. Setup.......................................................................................................................4C. Precautions ............................................................................................................5

2. Product Components.........................................................................................6

3. Hardware Overview ..........................................................................................7

4. Touch Screen Basics ..........................................................................................8A. Buttons and Icons on the Instrument Control Screen.....................................8B. Buttons on the Read Screen ................................................................................9C. Buttons on the Results Screen ..........................................................................10D. Buttons on the Tools Screen..............................................................................11E. Terminology Used in Parameter Settings.......................................................12

5. Home Screen .....................................................................................................13

6. Protocol Management .....................................................................................14A. Defining a New Protocol Using the Instrument Control Panel ..................14B. Modifying a Saved Protocol .............................................................................16C. Using the Protocol Composer to Set Up Multiple Read Formats...............17

7. GloMax-Multi+ Detection System with Shaking...................................19A. Selecting Shaking Options ................................................................................20

8. GloMax-Multi+ Detection System with Temperature Control............21A. Activating the Temperature Control...............................................................21B. Setting Incubation Parameters Followed by Plate Read ..............................22

C. Setting Incubation Parameters without Plate Read ......................................23

9. Plate Format Selection ....................................................................................24A. Changing the Plate Adapter Orientation .......................................................24B. Well Selection......................................................................................................25C. Selecting Reference Wells (Absorbance Mode Only) ...................................26D. Changing the Optical Crosstalk Mask for 384-Well Plates ..........................27E. Well Scan Mode .................................................................................................29F. Viewing Multiple Read Results for 6- to 48-Well Plates ..............................30G. Accessing the Interior of the Instrument ........................................................30

GloMax-Multi+ Detection SystemAll technical literature is available on the Internet at: www.promega.com/tbs/

Please visit the web site to verify that you are using the most current version of thisTechnical Manual. Please contact Promega Technical Services if you have questions on use

of this system. E-mail: [email protected]


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17. Detection Module Installation ......................................................................56A. Luminescence Module.......................................................................................56B. Fluorescence Module .........................................................................................57C. Absorbance Module ...........................................................................................58D. UV-Vis Absorbance Module .............................................................................61

18. Injector System Installation and Operation...............................................63A. Injector System Components ............................................................................63B. Installation Procedure........................................................................................64C. Injector System Operation.................................................................................67D. Using the Dispense Option ...............................................................................69

19. Tools ...................................................................................................................70A. System Information ............................................................................................71

B. Event Log.............................................................................................................72C. Sound Control .....................................................................................................72D. Setting the Time and Date.................................................................................72E. Updating Software .............................................................................................72F. Updating Firmware............................................................................................73G. External PC Control ...........................................................................................73

20. Maintenance......................................................................................................74A. General Instrument Care...................................................................................74B. General Cleaning ................................................................................................74C. Touch Screen Care..............................................................................................75

D. Removing and Reinstalling the Microplate Sample Tray Cover ................75E. Cleaning the Injectors ........................................................................................78F. Cleaning the Waste Collection Tray ................................................................78G. Replacing Injector Tips ......................................................................................79H. Inserting Injector Tip Assembly .......................................................................80I. Removing or Replacing Inlet and Outlet Plastic Tubing .............................80

J. Removing or Replacing Stainless Steel Tubing ........... ............ ............ ..........80K. Changing the Standard Light Plate Battery ...................................................81

21. Troubleshooting...............................................................................................82

A. Table of Error Messages ....................................................................................82B. Table of Common Problems .............................................................................85

22. Appendix ...........................................................................................................87A. Specifications.......................................................................................................87B. Warranty and Service ........................................................................................89C. Certificate of Decontamination ........................................................................90D. Related Products.................................................................................................91

1. Description

The GloMax-Multi+ Detection System is an expandable multimode readerwith unbeatable performance. Each detection mode has dedicated optics for the



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highest versatility without sacrificing performance. The GloMax-Multi+Detection System can be used as a reader dedicated to a single mode or as amultimode reader. As your application needs expand, the system can easily

accept the add-on modules, which offer additional detection modes. Thisflexibility allows you to customize the system to fit your laboratory needs.

1.A. Inspection

Upon receiving the GloMax-Multi+ Detection System, inspect it carefully forany damage to the exterior such as scratches and/or dents. Make certain allaccessories are included. Refer to the checklist shipped with the instrument fororder-specific items. Save all packaging materials, if possible, in case theinstrument needs to be returned for service.

Figure 1. The complete GloMax-Multi+ Detection System. A. GloMax-Multi+Instrument. B. Power cord. C. Power Supply Brick, 24V, 150W. D. USB Flash Drive,2GB. E. Fluorescence Optical Kit (included with Fluorescence Module).F. Luminescence Standard Light Plate (optional). G. Non-sterilized black or white96-well plate. H. Waste Collection Tray (included with injector system). I. OutletInjector Tube Assembly (included with injector system).J. DB-15 Serial Cable(included with injector system). K. Injector System (optional). L. Two Optical WellCrosstalk Masks. M. Microplate Adapter.

1.B. Setup

1. Place the GloMax-Multi+ Detection System on a flat, level surface. Leaveat least 7.5 inches (19cm) of clearance in front of the instrument to allow theinstrument door to open without hindrance. Position the instrument so thatthe touch screen faces outward.

2. Manually open the instrument door and remove the foam packing from theinside of the instrument. The packing material is used to secure the opticalheads and Microplate Sample Tray during shipping.

Caution: Remove all of the foam packaging from inside the instrumentbefore initial power up. Failure to do so may damage the electronics.

Promega Corporation 2800 Woods Hollow Road Madison, WI 53711-5399 USAToll Free in USA 800-356-9526 Phone 608-274-4330 Fax 608-277-2516 www.promega.comPart# TM311 Printed in USA.Page 4 Revised 5/10

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3. Plug the power cord into the power connector on the back panel of theinstrument (Figure 3). Plug the power supply brick into a power outlet. Forpower specifications, refer to Section 22.A.

4. Turn on the GloMax-Multi+ Detection System. The power switch islocated on the back panel next to the power connector.

5. Look for an LED light to come on within one minute. The light is located tothe left of the touch screen and indicates when the instrument is initialized.

6. After warming up, the touch screen will activate and default to the Homescreen.

If the optional injector system (for use with 6- to 96-well plates only) wasincluded with the purchase of the instrument, refer to Section 18 for

installation and operation instructions.

1.C. Precautions

Keep the instrument door closed when access to the interior of the instrumentis not needed. Leaving the door open for an extended period of time willresult in damage to the Photomultiplier Tube (PMT) due to light exposure,dust accumulating on the mechanical parts, or physical damage due toaccidentally bumping the open door.

Important: Close the instrument door when the instrument is not in use.

The GloMax-Multi+ Detection System is intended for indoor use only. Cleanup any spills immediately. The reader contains sensitive optical componentsand precision-aligned mechanical assemblies. Avoid rough handling.

Do not overfill microplate wells as this may lead to spills and/or damage thegaskets of the Microplate Sample Tray Cover. The sample residue can causethe optical head to malfunction. If any moisture appears on the MicroplateSample Tray Cover, clean the optical lens, the mask and the interior of theinstrument. See Section 20 for general care and cleaning instructions.

Note: Turn the instrument off any time you access the interior, install detection

modules or clean the injectors.

Warning: If injectors are installed, do not use bent or damaged injector tips.Check the tips periodically and replace as needed to avoid leaks onto theMicroplate Sample Tray Cover.


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2. Product Components

Product Size Cat.#

GloMax-Multi+ Detection System Base Instrument

with Shaking 1 each E8031

GloMax-Multi+ Detection System Base Instrumentwith Heating and Shaking 1 each E9031

Cat.# E8031 and E9031 must be purchased with at least one detection module(Cat.# E8041, E8051, E8061 or E9061; below).

Cat.# E8031 includes:

1 GloMax-Multi+ Base Instrument with Shaking 1 Power Cord 1 Power Supply Brick, 24V, 150W

1 USB Flash Drive, 2GB 2 Optical Crosstalk Masks 1 Plate Adapter 1 Protocol (on CD)

Cat.# E9031 includes:

1 GloMax-Multi+ Base Instrument with Heating and Shaking 1 Power Cord 1 Power Supply Brick, 24V, 150W 1 USB Flash Drive, 2GB 2 Optical Crosstalk Masks 1 Plate Adapter 1 Protocol (on CD)

Available Separately

Product Size Cat.#

GloMax-Multi+ Luminescence Module 1 each E8041

GloMax-Multi+ Fluorescence Module 1 each E8051

GloMax-Multi+ Visible Absorbance Module 1 each E8061

GloMax-Multi+ UV-Visible Absorbance Module 1 each E9061



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4. Touch Screen Basics

The touch screen is sensitive to the light pressure of a fingertip. It is notnecessary to use a stylus. To select a function, touch the corresponding button

once. Do not touch the LCD screen with any sharp object, pen, pencil, ormarker as these may cause permanent damage. Avoid spilling any liquid ontoor near the touch screen.

A sleep mode is initiated after 15 minutes with no activity or user stimulationof the touch screen. Lightly touch the LCD screen once to reactivate it.

4.A. Buttons and Icons on the Instrument Control Screen


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Home Screen.

7027TA

Help Screen.

7028TA

Opens and closes the instrument door. Changes to a Stop button when a run is in

progress. Touch the Stop button to cancel a run, if needed.

7029TA

Selects one of the three screens in the InstrumentControl Screen.

7030TA

Instrument Status bar (above).

7031TA Plate icon, visible when a microplate or a Waste

Collection Tray is detected inside the instrument. Icon turns dark when a microplate has been read.

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Detection Mode icon. A gray icon indicates whichdetection module is installed.

A black icon indicates the active module.

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Indicates the instrument is ready to begin a run.

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Indicates one or two injectors are installed.

7992TA

Temperature Display icon. Indicates that theheater option is activated.

Instrument temperature displayed to the right.

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Visible when a USB flash drive is detected. Icon turns dark when files have been saved to the

USB flash drive.


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4.B. Buttons on the Read Screen


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Protocol button. Touch the white text button to see the list of

preset and user-defined protocols.

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Plate Mapping button and well selectionindicator.

Touch the button to specify which wells are to beread.

Green wells are selected for reading. Gray wells are deselected and will not be read.

7036TA In Luminescence mode, this button enables

Kinetic Mode.

7037TA In Absorbance mode, the Wavelength button

enables single or dual wavelengths.

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Starts a run.

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Activates the PMT before starting a luminescencerun.

PMT activation is required before the firstluminescence run of the day.

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The Setup button appears when injectorsare connected.

Prepares the injector(s) for a prime, a flush and areverse purge.

7041TA

Touch one of the Injector buttons to activate theinjector(s).

Green indicates the injector is selected. Gray indicates the injector is deselected.

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Button for keypad entry.

Used in the User Protocol screen to enter aprotocol name.

7919TA Incubation button.

7920TA Shaking button.

7921TA Dispense button.


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4.B. Buttons on the Read Screen (continued)

4.C. Buttons on the Results Screen


7044TA Displays a list of the 50 most recent results files.

7045TA Use the Up and Down buttons to scroll

through the 50 most recent results.

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Use the Right and Left arrow buttons toscroll the wells in columns 112 for 96-wellplates or 124 for 384-well plates.

7047TA

When viewing Absorbance results, the buttontoggles between the different data formats: OD,%T and %A.

7048TA

Toggles through the sets of results generatedfrom repeat runs.

7993TA

Plate View button shows the full or partialview of the results in a multiwell format.

7050TA Displays the results as a ratio.

See Section 15 for ratio calculations.

7051TA Copies results files in .csv format to a USB

flash drive.

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Delete files and delete protocols button.

7922TA

User Prompt button. Interrupts an assay sothat instrument parameters can be adjusted.

7923TA Single measurement for each assay well.

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Well scan mode for plate formats other than 96-and 384-well standards. Allows for ninemeasurements within 6-well plate format.Allows for five measurements within 12-, 24-and 48-well plate formats.


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4.D. Buttons on the Tools Screen


7052TA Displays instrument-related information.

7051TA

Transfer instrument event log in .txt formatthrough the USB port to a USB flash drive.

7053TA Sound on/off.

7054TA Set time and date.

7055TA

Port setting for PC Connectivity.

Visible on the PC version of the software.

7056TA

Use to update software versions.

7057TA

Switches control of the instrument to a PC. Once switched, the PC has full control of the

instrument and stores all the data collected onthe PC hard drive.


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4.E. Terminology Used in Parameter Settings

Delay: The number of seconds between injecting the reagent and taking ameasurement. Setting a delay after the injection will allow flash-typeluminescence to fully actualize before a reading. The minimum delay value is

0.5 seconds in 0.1-second increments.

Dispense: The injector system delivers 25200l of reagent in 5l increments.This option can be used in conjunction with the Incubation icon to allowsufficient incubation time in the plate.

Flush: The protocol used to clean the injector system after a run. Therecommended flush protocol consists of steps for washing with deionizedwater, 70% ethanol, deionized water and then air-drying.

Integration: The duration of measurement per well ranging from 0.1 to 10seconds in 0.1-second increments. In assays where noise is much lower than

signal, a 1-second integration time will yield the same sensitivity as a 10-second integration time. Unless specified by the assay protocol, a 1-secondintegration time should be sufficient.

Optical Kit: Selects a Fluorescence Optical Kit from a list.

Period: The time interval between readings of the same well ranging from 1 to120 minutes in 1-minute increments.

Preset Protocols: A set of popular assay protocols preloaded into theinstrument for user convenience. These protocols cannot be renamed ordeleted. A user-modified version can be saved by touching the User button

on the Protocol screen.Prime: The injector and tubing are rinsed and filled with the reagent.

Protocol Composer: The section of the Graphical User Interface (GUI) to whichthe Protocol Options are dragged. This defines the order in which the protocolwill run.

Protocol Options: The list of available instrument operations that can beselected and dragged into the Protocol Composer.

Reading: The number of times a sample plate is read. The number of reads canbe set between 1 and 99.

Reverse Purge: The protocol retrieves any unused reagent from the injectorsystem. After a run, this protocol can be used to push the reagent back into thereagent bottle to reduce waste.

User Protocols: Protocols created by the user, which can be retrieved later. Allparameters including the plate well mapping are saved. These protocols can bemodified, renamed or deleted.

Volume: The volume injected into each well ranges from 25200l in 5lincrements for 6- to 96-well plates. Determine the sample volume per wellbefore selecting an injection volume. Although the maximum volume of a

typical 96-well plate is 300l, the recommended maximum volume per well foruse with this instrument is 250l. Overfilling a well will result in spills andpossible damage to the instrument.

Wavelength: Selects one or two Absorbance filters from a list.



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5. Home Screen

Figure 4. Home Screen. The Home screen contains four options:

New Protocol: Launches the wizard to set up a new protocol. Select Protocol: Launches the wizard to select preprogrammedprotocols from a list.

Instrument Control: Instrument setup, managing results andmanaging the instrument.

Help Topics: Covers informational topics.

From any of the Protocol wizards, return to the Home screen by touching theCancel button. In the Instrument Control screen, use the Home buttonfound at the bottom left of the touch screen.

Figure 5. Home button.


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7. To save the modified protocol:

a. Touch the white text button with the protocol name.b. Touch the Yes button in the dialog box.

c. To save the protocol under the same name, touch the Save button.d. To save the protocol under a new name, touch the Edit button. Usethe activated keypad to enter the new name and touch the OKbutton when editing is complete. Then touch the Save button to savethe renamed protocol.

e. Modified protocols are saved in the User list. See Figure 9.

To delete a protocol from the User list, highlight the protocol, then touch theDelete button. Protocols from the preset list cannot be deleted.

6.C. Using the Protocol Composer to Set Up Multiple Read Formats

The Protocol Composer on the GloMax-Multi+ System allows multiple assayrequirements to be strung together into one protocol. Simply select theparameters of choice by dragging the appropriate icon from the ProtocolOptions list section on the left to the Protocol Composer section on the right.Once the Protocol Options icons have been selected, the user may definespecific requirements for each option in the window on the right.

Figure 10. Dragging icons from Protocol Options to the Protocol Composer.

1. Select and drag Protocol Option icons onto the Protocol Composer(Figure 10).


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6.C. Using the Protocol Composer to Set Up Multiple Read Formats (continued)

2. With the Protocol Option icons chosen, define the desired plate layout andOptical Kit.

Note: Once a plate format is selected, this formatting will become thedefault for all subsequent selections.

Figure 11. Defining a multiple parameter protocol.

Figure 12. Defining user prompt message screen.


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3. Select and drag the User Prompt icon to the Protocol Composer andtype an instructional message into the Edit bar (Figure 12).

4. Select and drag the Fluorescence icon to the Protocol Composer and

select the Red Optical Kit (Figure 13).

5. Insert a plate into the GloMax-Multi+ Detection System and press theStart button to begin a run.

Figure 13. A Defined Protocol ready to run.

7. GloMax-Multi+ Detection System with Shaking

The GloMax-Multi+ Detection System with Shaking allows the sample plate tobe shaken as is necessary to meet the requirements of certain assays. Selecting

the Shaking icon allows one to define the parameters of shake operation suchas time, intensity and type of motion for the shaking pattern.

Time: The GloMax-Multi+ Detection System with Shaking allows for shakingintervals of 0.1120 minutes.

Intensity: The GloMax-Multi+ Detection System with Shaking has three ratesof defined shaking intensity: low, medium and high. These settings roughlyequate to 150, 300 and 500 rpm, respectively.

Type of motion: Either linear, where the entire platform of the shaker moves ina linear motion or orbital, where the entire platform of the shaker moves in acircular orbit.


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7.A. Selecting Shaking Options

1. Select and drag the Shaking icon from the Protocol Options to theProtocol Composer (Figure 14).

2. Touch the Time button to select the duration of shaking.

3. Touch the Intensity button to select the rate of shaking desired.

4. Touch the Type button to select the pattern of shaking desired.

Important: You must use a cover film or plate seal on the plate to preventspillage and loss of sample when using the plate shaking option. If using low-density plates with shaking, it is necessary to use a cover film to preventspillage and loss of sample.

Figure 14. Selecting the Shaking parameters.

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8. GloMax-Multi+ Detection System with Temperature Control

A new feature found in the GloMax-Multi+ Detection System, along with theshaker (Section 7) is the plate heating device. The GloMax-Multi+ Detection

System Heater is capable of heating from ambient 20C to 45 0.75C. Before arun, allow ten minutes for the heater to warm up and calibrate to the desiredtemperature. Access the Temperature Control option by selecting the Toolsbutton on the Instrument Control Screen. After two hours of inactivity, theTemperature Control option will automatically shut off. If the temperature ofthe instrument is different than the desired temperature when an assay run isstarted, the GloMax-Multi+ Detection System will display a warning windowwith the option to continue with or cancel the run.

Important: The use of a plate cover film is required to prevent sampleevaporation and damage to the instrument.

8.A. Activating the Temperature Control Option

1. Touch the Tools button to go to the Tools screen (Figure 15).

2. Touch the Activate Heater button to select the heater option.

3. Enter the desired temperature using the keypad.

4. Wait at least 10 minutes for the heater to calibrate to the desired

temperature.

Figure 15. Accessing the Heater option via the Tools button.

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8.B. Setting Incubation Parameters Followed by Plate Read

1. Activate the Heater option (Section 8.A).

2. Select and drag the Measurement icon from Protocol Options to theProtocol Composer and define the measurement parameters.

Figure 16. Setting the Incubation Parameters followed by Plate Read.

3. Select and drag the Incubation icon from Protocol Options to the ProtocolComposer (Figure 16).

4. Touch the Time button to select the length of incubation.

5. Touch the Temperature button to set the desired assay temperature indegrees Celsius (C).

6. Touch the Temperature Control button to activate the Heater controls.

7. Touch the Door button to automatically open the instrument door.

8. Place the sample plate on the Microplate Sample Tray. Well A1 must be atthe top right corner.

9. Touch the Door button again to retract the sample plate back into theinstrument.

10. Press the Start button.


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8.C. Setting Incubation Parameters without Plate Read

Figure 17. Setting the Incubation Parameters without Plate Read.

1. Activate the Heater option (Section 8.A).2. Select and drag the Incubation icon from the Protocol Options to the

Protocol Composer (Figure 17).

3. Touch the Time button to select the length of incubation.

4. Touch the Temperature button to set the desired assay temperature indegrees Celsius (C).

5. Touch the Temperature Control button to activate the Heater controls.




9. Press the Start button.

Note: For assays that require an incubation step without heat, omitactivating the Heater option on the Tools screen and do not select the

Temperature Control button.


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3. Firmly snap the plate adapter into place to continue with an assay run.

Caution: The Microplate Sample Tray should only have the cover in place for96-well plate formats. For all other plate formats, the cover must be removed.

For instructions on how to remove the Microplate Sample Tray Cover refer toSection 20.D.

9.B. Well Selection

To select or deselect a well, touch the well. This will toggle the coloration ofthe well on the touch screen. Green indicates that a well is selected and grayindicates that a well is deselected. In Absorbance mode, touching the screenalso toggles to Ref, which refers to reference wells.

To select or deselect a whole row or column, touch the corresponding number

or letter. Hold down the button and drag to select multiple rows or columns.When the All button is touched, the entire plate toggles between being eitherselected or deselected.

1. Touch the Plate button to open the Plate Mapping screen. Select wells tobe read. Green indicates a well is selected. Gray indicates a well isdeselected (Figure 19).

2. The message box below the wells summarizes the well selection.

3. Touch the OK button to commit changes made on the Plate Mappingscreen. Touch the X button to cancel any changes made.

4. The wells that are selected to read are saved with each protocol setting.When using a saved protocol, always double-check that the well selectionis properly set to read the intended wells.

Figure 19. Plate Mapping screen. On the plate mapping screen, green indicatesselected wells, gray indicates deselected wells.

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9.C. Selecting Reference Wells (Absorbance Mode Only)

Reference wells are optional in absorbance mode. The detection mode needs tobe set to Absorbance to enable this feature. Touch a well to toggle between

selected, deselected and reference (Figure 20). Reference wells must be selectedindividually. It is not possible to select by column or row.

Readings taken from Reference wells are used as a blank. If more than onewell is selected as a Reference, an average of the values from these wells isapplied to each of the readings.

Figure 20. Selecting Reference wells (Ref), as seen in Absorbance mode.

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9.D. Changing the Optical Crosstalk Mask for 384-Well Plates

Perform these steps only when using the 384-well plate format.

1. Remove the injector tips from the injector tip holder.2. Remove the injector tip holder from the optical head. Facing the front of

the GloMax-Multi+ Detection System, push the optical head upward, thenleftward (Figure 21).

Figure 21. Changing the optical crosstalk mask.

3. Place your hand underneath the optical mask. Grasp the sides of the opticalcrosstalk mask and pull it towards you (Figure 22).

Figure 22. Grasp the sides of the optical mask and pull it towards you, beingcareful to not touch the bottom of the mask.

Note: Do not touch the bottom of the optical head.

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9.D. Changing the Optical Crosstalk Mask for 384-Well Plates (continued)

4. Replace the 96-well optical crosstalk mask by sliding the 384-well opticalcrosstalk head mask into place on the optical head. Align the mask with the

optical head and gently guide the optical crosstalk mask onto the opticalhead (Figure 23).

Figure 23. Slide the 384-well optical crosstalk mask into place on the optical head.

5. Return the injector tip holder to the optical head. Position the holder justabove the front pins on the optical head. Push the holder toward the rightto lock the holder securely to the right pin of the optical head. Then pushdown and lock the holder on the left pin (Figure 24).

6. Insert the injector tip(s) into the injector tip holder and snap into place(Figure 25).

Figure 24. Slide the 384-well optical crosstalk mask into place on the optical head.

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9.F. Viewing Multiple Read Results for 6- to 48-well Plates

In lower density formats where multiple readings are taken, the results areshown as an average for each well. To view the results of readings for a

particular well, zoom in on the well and touch the well on the User interfaceonce the plate read is finished.

Figure 27. Viewing Multiple Read results per well for 6- to 48-well plates.

9.G. Accessing the Interior of the Instrument

Touch the Door button to open and close the instrument door.The Microplate Sample Tray automatically extends and retractswhen the Door button is touched. If the 96-well MicroplateSample Tray Cover is installed, it will also automatically open andclose with the Microplate Sample Tray. Do not use force to close the

Microplate Sample Plate Cover.Caution: Do not use force to close the Microplate Sample Plate Cover. Thecover and instrument door will automatically close after the Door button istouched.

Opening the instrument door while the Microplate Sample Tray is in motionwill cause it to stop. When it is necessary to access the instrument interior,turn off the instrument. Manually hold the instrument door open. Do not usethe Door button. The Microplate Sample Plate Cover may be in the way.

Important: Close the instrument door immediately after each use.


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10. Luminometer Operation

10.A. General Information

The Luminometer detection wavelength range is 350 to 650nm.Microplate Recommendations

White plates are recommended for general luminescence detection. Black plates are recommended primarily for bright assays, such as

ELISA, but generally will yield decreased sensitivity despite lowerbackground and crosstalk than white plates.

Clear plates are not recommended due to high crosstalk.

Luciferase assay samples and reagents must be equilibrated to roomtemperature prior to taking measurements to ensure optimal light intensity.

For best results, use freshly prepared reagents and follow assay protocolscarefully.

To reduce background noise, keep the interior of the instrument clean. Toreduce crosstalk from adjacent wells, always use the Microplate Sample TrayCover. Flush the injectors after each use and clean up any spills. See Section 20for detailed cleaning instructions.

10.B. Microplate Sample Tray Cover

The Microplate Sample Tray Cover is installed on the Microplate Sample Tray

as part of the Luminescence module. Its primary purpose is to reduce crosstalksignal from adjacent wells.

The Microplate Sample Plate Cover is for use only with 96-well plates. Thecover must be removed prior to running any other plate format. SeeSection 20.D for Microplate Sample Plate Cover removal.

If any moisture appears on the Microplate Sample Tray Cover, clean theoptical lens, the mask and the interior of the instrument. See Section 20.Bfor general care and cleaning instructions.

10.C. PMT Activation

The Luminescence module uses Photomultiplier Tube (PMT) detection. Itrequires a 5-minute warm-up to ensure consistent results. Activating the PMTprior to preparing the sample plate for reading will reduce the wait timebefore the run.

When the Luminescence mode is selected, an Activate PMT button is visiblein place of the Start button. This reminds you to warm up the PMT beforeinitiating a run. After PMT activation begins, the Start button appears.


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10.C. PMT Activation (continued)

While the PMT is activating, you can maneuver around the software and selectother detection modalities without interfering with the PMT activation.

However, if a run is started in a different detection mode or the instrumentdoor is open for more than one minute, the PMT will deactivate.

Parameters and file names can be set while the PMT is activating. If the Startbutton is touched before the PMT is completely activated, the run will beginimmediately after the 5-minute warm-up of the PMT.

10.D.Starting a Run

Easy-to-use protocol wizards can be accessed from the Home screen to set upthe run parameters. See Section 6 for protocol management. Otherwise, follow

the steps below to set up a Luminescence run.1. From the Home screen, touch the Instrument Control button, then the

Read button (Figure 28).

2. Select and drag the Luminescence icon from the Protocol options to theProtocol Composer.

3. Touch the Plate Format button and select the plate format that isrequired.

4. Touch the Plate button and select the wells to be read. Green indicates a

well is selected and gray indicates a well is deselected.

Figure 28. Defining a Luminescent Plate read.

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5. If injectors are used, press the Injector icons to activate injectors and setthe Volume, Delay and Integration time.


7. Place the sample plate on the Microplate Sample Tray. The A1 well must beat the top right corner.

8. Touch the Door button again to automatically close the MicroplateSample Tray Cover and retract the sample plate back into the instrument.Do not push down on the Microplate Sample Plate Cover or force it toclose.

9. Touch the Start button.

10. To name the results file, touch the Edit button to activate a keypad. Enter

the results file name and touch the OK button when editing is complete.

11. Touch the OK button on the dialog screen to save. The GloMax-Multi+Detection System will initialize and begin the run.

12. To stop a run at any time, touch the Stop button.

13. Remove the sample plate after the run is complete.

Note: To prevent dehydration and spills, remove the sample plate from theinstrument when a reading is completed.

10.E. Kinetic Mode

In Kinetic mode, the instrument performs multiple reads of a single well withdefined frequency over a defined period of time before moving to the nextwell. The kinetic feature is available in Luminescence mode only. One or bothinjectors can be used in the Kinetic mode. Each sample reading is taken afterthe second injection and injection delay.

Note: Kinetic Mode is not compatible with the 384-well plate format or WellScan Mode for 6- to 48-well plate formats.

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10.F. Kinetic Parameters

Figure 29. Setting Kinetic Parameters. Arrow points to Kinetic button.

Initial Delay: Delay before the start of the first reading to allow the sampleplate to dark-adapt. The range is 060 minutes and can be adjusted in1-minute increments.

Kinetic Interval: The delay between the last reading of the previous welland the first reading (or first injection) of the next well. The range is 0.160seconds and can be adjusted in 0.1-second increments.

Plate: Shows the time estimated to read the entire sample plate using thedefined parameters.

Readings: The number of data points to collect per well. The range is 2250 in1-unit increments.

Well: Shows the time taken to read each well using the defined parameters.

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10.G.Running the Standard Light Plate Protocol

The optional Standard Light Plate provides a quick way to verify instrumentperformance. The Standard Light Plate consists of three highly stable light

sources that simulate luminescent samples at signal intensity ranging overfour logarithmic scales. Refer to the instruction card enclosed with theStandard Light Plate for detailed instructions on use. The Standard Light Plateprotocol is available as a preset protocol.

Note: See Section 15 for data analysis details.

Getting Started

1. Briefly press the Start button on the Microplate Luminescence LightPlate.

2. The Battery check indicator will flash a green light as long as the Startbutton is pressed and the battery has sufficient power.

3. The green light will turn Off once the Start button is released. The lightplate is now On and ready for use. After five minutes it will automaticallyturn Off. The built-in timer can be restarted at any time by pressing theStart button again.

4. If the green light does not appear while the Start button is pressed,replace the battery. See Section 20.K for details.

Running the Luminescence Light Plate Protocol5. From the Home Screen, choose Instrument Control.

6. From the Instrument Control screen, select Luminescence by pressing theDetection Mode button.

7. After Luminescence has been selected, press the Luminescence Protocolbutton and scroll down to Luminescence Light Plate.

8. Load the Luminescence Light Plate and press the Start button.

Analyzing the Results from the Luminescence Light Plate Protocol

9. Transfer the data file from the GloMax-Multi+ Detection System to anexternal computer via use of the USB flash drive.

10. Open the .csv file with Microsoft Excel and plot the results against 1 108

for B2, 1.2. 106 for D2 and 1 104 for F2.

11. The R2 value generated should be >0.98 (Figure 31). If the results are


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10.G.Running the Standard Light Plate Protocol (continued)

Figure 30. Viewing luminescence light plate results.

Figure 31. Calculating R2 value.

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Luminescence Light Plate Protocol

R2= 0.9999

0

1.0 107

2.0 107

3.0 107

4.0 107

5.0 107

6.0 107

7.0 107

8.0 107

0

2.E+

07

4.E+

07

6.E+

07

8.E+

07

1.E+

08

1.E+

08

1.E+

08

Luminescence(RLU)


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11.C. Starting a Run

Easy-to-use protocol wizards can be accessed from the Home screen to set upthe run parameters. Follow the steps below to set up an Absorbance run.

Figure 34. Setting up a single Absorbance Read.

1. From the Home screen, touch the Instrument Control button, and thenthe Read button.

2. Select and drag the Absorbance icon from the Protocol Options to theProtocol Composer.

3. Touch the white text button to select a protocol or modify the displayedparameters.

4. Single- or dual-wavelengths can be selected by toggling the Wavelengthbutton (Figures 34 and 35).

5. Touch the Wavelength Filter Selection button to select a wavelength.There is an option for each of the installed filters. For custom filters, selecteither position A or B.

6. Touch the Plate button and select wells to be read. Green indicates a wellis selected, gray indicates a well is deselected and Ref is for reference.



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11.C. Starting a Run (continued)

Figure 35. Setting up a dual absorbance read.



11. To name the results file, touch the Edit button to activate a keypad. Enterthe results file name and touch the OK button when editing is complete.

12. Touch the OK button on the dialog box. The GloMax-Multi+ DetectionSystem will automatically begin reading samples.

13. To interrupt a reading at any time, touch the Stop button.

14. Remove the sample plate after the reading is complete.

Note: To prevent evaporation and spills, remove the sample plate from theinstrument when the reading is complete.


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11.D.Ratiometric Assay

For ratiometric assays, two wavelengths can be selected for each well reading.The results screen can display only the OD values for each wavelength (two

values for each well) or a ratio of the two readings (when the Ratio button isselected). For more details on absorbance results, see Section 15.

12. UV-Vis Absorbance


The GloMax-Multi+ Detection System with UV-Vis Module detection has awavelength range of 200800nm. The system includes four fixed filters andtwo customizable filter holders for the 260nm (Paddle A) and 280nm

(Paddle B) filter paddles that are included in the six-position filters wheel. Twoabsorbance wavelengths can be selected per well when running ratiometricassays.

Microplate Recommendations

For Visible Region measurements, black-walled and clear flat-bottommicroplates are recommended. Clear microplates can also be used.

For UV-Vis region measurements, UV transparent plates(Costar Cat.# 3635) are recommended.

Note: When using clear microplates, we recommend using the Microplate

Sample Tray Cover to reduce light scatter from adjacent wells. See Section20.D for removing and attaching the Microplate Sample Tray Cover.

Warning: The Xenon Flash lamp emits ultraviolet light at levels that can injurethe eye. Do not open the instrument door while taking measurements.

12.B. Optical Filters Wheel

The Optical Filters Wheel located on the bottom of the Absorbance DetectionHead (Figure 32) holds four fixed optical filters and two customizable filterpaddles. The wavelength is easily selected by using the touch screen interface;

the appropriate filter is automatically selected by the instrument. A manualchange of the filter is not required.


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12.B. Optical Filters Wheel (continued)

Table 2. Optical Wavelengths for Various Assays.


Filter Wavelength (nm) Assays260 Nucleic acid quantification

280 Protein quantification

450 ELISA

560 BCA Protein Assays

600Bradford Protein Assays, Coomassie Blue ProteinAssays

750 Lowry Protein Assays

12.C. Customizable Absorbance Paddles with 260nm and 280nm Filters

Two Absorbance Paddles are included for customization; 260nm and 280nmfilters come already installed for instruments ordered with the UV-VisAbsorbance Module (Cat.# E9061). The Absorbance Paddle can accommodateany 12.7mm in diameter and < 6.4mm thick filter, in the spectral range of200800nm. Contact Promega for the most current information on filteravailability.

Wavelengths for custom filters cannot be manually entered by the user. Youhave the option of selecting from the custom positions labeled A or B.

To assemble the Absorbance Paddle, simply place the filter (with the mirrorside down) into the opening. Press down on the filter collar until the filter fitssnugly. Avoid getting smudges or fingerprints on the filter.

To remove the filter, turn the Absorbance Paddle upside down. Press down onthe collar of the filter. Avoid touching the filter itself.

Figure 36. Absorbance filter paddle.

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13. Fluorometer Operation


Microplate Recommendations

Black plates are highly recommended. Clear plates can be used as a low-cost alternative if a high level of signal

intensity is expected. White plates are not recommended. They will increase noise by 2050

times.

Notes:

1. Samples with bubbles or high surface tension will affect fluorescencereadings.

2. Ensure the correct Optical Kit is used for the assay fluorophore.

3. If the instrument includes a Luminescence Module, it has a MicroplateSample Tray Cover installed. The Microplate Sample Tray Cover is notrequired for using fluorescence mode. To remove the Microplate SampleTray Cover, refer to Section 20.D for instructions.

4. All values recorded are in raw RFUs (relative fluorescence units). Tonormalize, use the Curve-Fitting Data Analysis Software.

13.B. Optical Wavelengths and Commonly Used Fluorophores

Table 3. Commonly Used Fluorophores.

*Optical kits are named based on the color of the excitation wavelength.

OpticalKits*

Excitation PeakWavelength

EmissionWavelength Typical Fluorophores

UV 365nm 410460nm Hoechst dye, 4-MU

Blue 490nm 510570nm EGFP, or hMGFP, PicoGreen,RiboGreen, Fluorescein, Quant-iT Protein, Quant-iTdsDNA, Rhodamine-110

Green 525nm 580640nm Rhodamine, Cy3, resorufin

Red 625nm 660720nm Cy5, Quant-iT RNA


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13.C. Starting a Run

Easy-to-use protocol wizards can be accessed from the Home screen to set upthe run parameters. Follow the steps below to set up a Fluorescence run.

1. From the Home screen, touch the Instrument Control button, and thenthe Read button.

2. Select and drag the Fluorescence icon from the Protocol Options to theProtocol Composer.

3. Touch the Plate Format button and select the plate format that isrequired.

4. Touch the Plate button and select the wells to be read. Green indicates awell is selected and gray indicates a well is deselected.

5. Select the Fluorescence Optical Kit.


7. Place the sample plate on the Microplate Sample Tray. The A1 well must beat the top right corner.

8. Touch the Door button again to automatically retract the sample plateback into the instrument.


Figure 37. Defining a fluorescence plate read.

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10. To name the results file, touch the Edit button to activate a keypad. Enterthe results file name and touch the OK button when editing is complete.

11. Touch the OK button on the dialog screen to save. The GloMax-Multi+

Detection System will initialize and begin the run.

12. To stop a run at any time, touch the Stop button.

13. Remove the sample plate after the run is completed.

Note: To prevent dehydration and spills, remove the sample plate from theinstrument when the reading is completed.

13.D.Switching Fluorescence Optical Kits

1. Close the instrument door by touching the Door button. This will ensure

the Microplate Sample Tray is at the Home position.2. Manually open the instrument door and hold it open with one hand.

3. Gently pull out the Fluorescence Optical Kit found under the Fluor label.

4. Insert a new Fluorescence Optical Kit into the opening by pushing it infirmly. The label should be facing up and outward.

Figure 38. Installing a Fluorescence Optical Kit.

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14. Repeat Runs

Each detection modality has the option to do repeat runs. Injectors cannot beused in this mode. In repeat runs, the entire sample plate is read between each

period.

Figure 39. Parameters for repeat runs.

14.A. Repeat Runs Parameters

Readings: The number of times a sample plate is read. When the number ofreadings is set to a value between 2 and 99, the period parameter and timeestimates are displayed. The time estimate for a plate (located below thereadings parameter) is based on the number of wells selected to read.

Period: The time difference between readings of the same well. If this value isless than the time estimated to read the plate, a caution message will appear toinform user that the actual interval between readings will be longer. Theinstrument will begin reading A1 (or the first well selected) immediately aftercompleting the last reading of the plate. The actual period time is indicated onthe results .csv file when exported to a computer. The range is between 1 to120 minutes in 1-minute increments.

14.B. Viewing Repeat Runs Data on the Results Screen

The results are displayed as a set of data on the Results screen. To view theother sets of data, touch the Plus button to move forward and touch theMinus button to move back. All data from repeat runs are saved in oneresults file. The GloMax-Multi+ Detection System saves all captured data toits internal memory. However, when the Files button is touched, only 50 ofthe most recent files are visibly listed. To view results files not displayed,touch the USB button to transfer all files to a USB flash drive and view themon a computer.


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Figure 40. Results screen with repeat runs data.

15. Data Management and Transfer

The GloMax-Multi+ Detection System saves all captured data to its internalmemory. However, when the Files button is touched, only the 50 most recentfiles are visible. To view other results files, touch the USB button to transferall the files to a USB flash drive and view them on a computer.

15.A. Viewing the Data in the Results Screen

Touch the files button to see a list of results files. The files are sorted by dateand time of the run. Select a file to view (Figure 41).

Figure 41. The Results screen of the instrument control screen. Touch the Filesbutton (upper right) to see a list of results files. The files will be sorted alphabeticallyand you can select a file to view.

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15.A. Viewing the Data in the Results Screen (continued)

Figure 42. Deleting data from the Results screen.


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Use the Up and Down arrow buttons to scrollthrough the displayed list of results files.

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The results are displayed in a microplate format in eithera full or partial view. Toggle between the two views bytouching the Plate View button between the horizontalarrows. A partial view of the results displays three or sixcolumns of data depending on the protocol.

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Use the Left and Right arrow buttons to scroll throughthe results in partial view.

Use the Up and Down arrow buttons to scrollthrough the wells in rows AH for 96-well plates or rowsAP in 384-well plates.

Use the Delete button to permanently removeunwanted results from the results list.


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15.B. Luminescence Data

Luminescence data is displayed in Relative Luminescence Units (RLU).

Kinetics

Kinetic results are displayed as a graphic curve in the Results screen. To obtainnumeric readings, export the results file to a USB flash drive and transfer it toa computer. See Section 15.F for transferring data. There will be a single resultsfile for the entire run.

The X axes are displayed in linear time, and Y axes are displayed in log RLUscale. The Y axes have a minimum range of 1 log (a result value range of 10).Each well auto-adjusts to display the graph in full Y-axis scale.

Ratio Readings

When two injectors are used in a protocol, two readings are taken for eachsample. Results are displayed side-by-side in the corresponding well (partialview) or first reading on top of the second reading (full view). To view a ratioof the two readings, touch the Ratio button. The ratio calculated forluminescence data is different than the ratio calculated for absorbance data.The luminescence ratio is calculated as:

Ratio =1st Reading2nd Reading

Note: A value of 1 1029 indicates saturation of the luminescence signal.

15.C. Absorbance Data

Single wavelength absorbance data can be displayed in three different units:Optical Density (OD), Percent Transmittance (%T) and Percent Absorbance(%A). Data exported to .csv format is only in OD units. To view the differentunits in the Results screen, toggle the button corresponding to the unit type.

Data calculations:

Transmittance (T) =I

I0

%T = 100T

OD = log10(A)

Absorbance = 2 log10(%T)

I is the light intensity at a specific wavelength that has passed through thesample. I0 is the light intensity before it enters the sample.

Note: The maximum OD value of 5.0 means there was zero transmittance.


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15.C. Absorbance Data (continued)

Wells marked with Ref were designated as Reference wells. The referencevalue of each wavelength reading is subtracted from each wavelength reading.If more than one Reference well is selected, an average of the Reference valuesat each wavelength is used.

Ratiometric Data

Figure 43. Absorbance ratiometric results as displayed on the Read screen.

When two wavelengths are selected for a protocol, a reading at eachwavelength is taken for each sample. The results are displayed in OD units inthree possible formats.

In partial view, reading 1 and reading 2 are displayed side-by-side in thecorresponding well (Figure 43, Panel A).

In full view, reading 1 is displayed above reading 2 in the corresponding

well (Figure 43, Panel B). When the Ratio button is touched, the results are displayed as a single

value (Figure 43, Panel C).

The ratio value calculated for absorbance data is different than forluminescence data. The absorbance value is calculated as:

Absorbance Value = 2nd reading 1st reading

15.D.Fluorescence Data

The Relative Fluorescence Units (RFU) are displayed with two significantdigits.

Note: A value of 1 1033 indicates saturation of fluorescence signal.

15.E. Multiplexing

The GloMax-Multi+ Detection System simplifies the process of multiplexingassays. When multiplexing, read the first assay based on the protocol neededfor the reagent. To multiplex, press the Multiplex button displayed in theResults screen, and select or generate the next detection protocol needed. The

resulting data will be saved together in a single file.


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15.F. Transferring Data

Results files are transferred to a computer via a USB flash drive using the USBport located to the left of the touch screen. The files are exported in a .csvformat and are compatible with Microsoft Excel on both Windows andMacintosh computers. Step-by-step directions follow.

1. To transfer data from the GloMax-Multi+ Detection System to an externalcomputer, insert a USB flash drive into the USB port located left of thetouch screen. When the USB flash drive is detected, an icon depicting aUSB flash drive should appear on the Instrument Status bar.

2. Touch the Results button on the Instrument Status screen.

3. Touch the Files button, and highlight the file(s) to be exported.

4. Touch the USB button under the Tools button.5. Touch the OK button to export only the selected file(s). Touch the Copy

All Files button to export all of the results files. The .csv formatted file(s)will be saved onto the USB flash drive in the folder named: Promega\Data

6. Move the USB flash drive to your local computer.

For Windows computers, double-click to open the file in Excel. For Macintosh computers, click to open the file and import the data into

Excel.

Most USB flash drives are compatible with the GloMax-Multi+ DetectionSystem. USB flash drives greater than 1GB may take longer than others to berecognized. Wait a few seconds for the instrument to recognize the USB flashdrive. If the USB flash drive is not detected after a few seconds, try a differentflash drive. Call Promega Technical Services for details regarding which USBflash drives are currently known to be incompatible.

Note: Results files can be transferred from the instrument to a computer onlyvia a USB flash drive. Use the USB port on the front of the instrument.

After results files are transferred, backup copies will remain on the internal

memory of the GloMax-Multi+ Detection System. The 50 most recent files aredisplayed in the Files window of the Results screen.

Due to the nature of the data generated by the GloMax-Multi+ DetectionSystem, it is necessary that computers used for data capture and analysis arecapable of reading data with a comma-separated value. To ensure best results,the computer should be operating in English (United States) mode. This optionis accessible in the Windows Control Panel in the Regional and LanguageOptions.

Note: Inserting the USB flash drive while the instrument is conducting a plate

read is not recommended. We recommend that you either insert it before orafter pressing the Start button.

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16. Connecting to a PC

For added flexibility, you have the option of connecting the GloMax-Multi+Detection System to a PC. See Section 22.D for ordering information. It is

possible to control the GloMax

-Multi+ Detection System remotely using anexternal PC. This allows you to store the results directly on the hard drive of aPC. Connecting directly to a Macintosh computer is not currently supported.


Install the necessary software, which is part of the GloMax-Multi+ DetectionSystem External PC Connect Kit (Cat.# E8071), before connecting the instrumentto a PC. The results files will not be saved on the internal computer of theGloMax-Multi+ Detection System when an external PC is connected. Data fromthe instruments internal computer can be transferred only through the USB port

located to the left of the touch screen. When an external PC has control of theinstrument, the GloMax-Multi+ Detection System touch screen is disabled.

16.B. System Requirements

Windows-based computer with Windows XP or Windows Vista

operating system Microsoft .NET Framework, version 2.0 (x86). Package is included with the

External PC Connect Kit USB cable or an RS-232 serial cable (Section 22.D) The PC version of the GloMax-Multi+ Detection System software

(included with the External PC Connect Kit, Cat.# E8071)

16.C. File Locations

While an external PC is controlling the GloMax-Multi+ Detection System,newly generated results files can be found in the following locations. Thelocation of the folders cannot be defined by the user. We recommend creatinga desktop shortcut link to the following folders.

Data files = C:\Documents and Settings\login name\MyDocuments\Promega\GloMax-Multi Plus\Results

Protocol settings = C:\Documents and Settings\login name\MyDocuments\Promega\GloMax-Multi Plus\Protocols

Event log = C:\Program Files\Promega\GloMax-Multi Plus\EventLog.txt



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16.D.Installing Microsoft .NET Framework version 2.0 (x86)

The following steps are necessary only if Framework software is not alreadyinstalled on the computer. Perform either step one or step two.

1. Insert the External PC Connect software CD into the PC. View the files onthe CD, and double-click on the file dotnetfx.exe. Follow the step-by-stepinstructions to install the software.

2. Alternatively, download .NET from this web site:www.microsoft.com/downloads/

16.E. Installing the Drivers for the GloMax-Multi+ Detection System

When connecting to a PC through a USB cable for the first time, the computerwill detect the instrument as a new device. The drivers necessary to connect to

the instrument are included with the PC version of the software. Follow theinstructions below to install the drivers. The following example is based onusing the Windows XP operating system.

1. Install the software onto the PC.

2. Connect the GloMax-Multi+ Detection System to the PC using the USBcable. Use the USB port on the back panel of the instrument. The USB porton the front of the instrument is used only for data transfer.

3. Turn on the GloMax-Multi+ Detection System.

4. The Found New Hardware wizard will appear. Follow the step-by-stepinstructions below to complete the wizard.

5. Select: No, Not This Time onthe Welcome screen, then selectthe Next button to continue.

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16.E. Installing the Drivers for the GloMax-Multi+ Detection System (continued)


6. When asked, "What do youwant the wizard to do?", select

the default option Install theSoftware Automatically. SelectNext to continue.

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7. The driver will begin to install.A warning message will appear

to inform the user that thedriver has not passed MicrosoftWindows Logo testing. SelectContinue Anyway.

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8. As the driver finishes installing,a status window will appear.

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9. Select the Finish button whenall of the installation steps havebeen completed.

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10. A message reading Your new hardware is installed and ready to use willappear on the Taskbar to confirm successful installation.


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16.F. Connecting to a PC Using a USB Cable

1. Turn on the instrument and computer.

2. Connect the GloMax-Multi+ Detection System to the computer using a

USB cable. Use the USB port on the back panel of the instrument. The USBport on the front of the instrument is used only for data transfer.

3. From the Home screen, go to the Instrument Control screen, and touch theTools button.

4. Touch the External PC Control button.

5. Launch the GloMax-Multi+ Detection System PC software from thecomputer.

6. Check the Instrument Status bar of the PC software for connectivity. Whenthe connection is successful, the Instrument Status bar should readReady.

7. If it does not, select the Tools button of the Instrument Control screen onthe computer, and confirm that the Instrument Port setting is on USB.

From this point, all instrument control should be done from the computer. Toreturn control to the GloMax-Multi+ Detection System, follow theinstructions in Section 16.H.

16.G.Connecting to a PC Using an RS-232 Serial Cable

1. Turn on the instrument and computer.

2. Connect the GloMax-Multi+ Detection System to a computer using theRS-232 serial port located on the back panel of the instrument.

3. From the Home screen, go to the Instrument Control screen, and touch theTools button.

4. Touch the External PC Control button.

5. Launch the GloMax-Multi+ Detection System PC software from the

computer.

6. Check the Instrument Status bar of the PC software for connectivity. Whenthe connection is successful, the Instrument Status bar should readReady.

7. If it does not, touch the Tools button on the Instrument Control screen ofthe computer, and confirm that an appropriate COM Instrument Port isselected.

From this point, all instrument control should be done from the computer. To

return control to the GloMax

-Multi+ Detection System, follow theinstructions in Section 16.H.


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17.B. Fluorescence Module

The Fluorescence Module is required for fluorescence detection. If yourinstrument does not have this module, contact Promega Technical Services for

information about purchasing and installation of the Fluorescence Module. SeeSection 22.D for ordering information.

Installing the Fluorescence Module

1. If the door is open, touch the Door button to automatically close theinstrument door and set the Microplate Sample Tray into Home position.

2. Turn off the GloMax-Multi+ Detection System.

3. Manually open the instrument door, and hold it open.

4. Slide the Fluorescence Detection Head into the position labeled Fluorlocated inside the instrument. Be careful not to damage the printed circuitboard on the back of the detection head.

5. Hand-tighten the two captive screws using the Allen Wrench (size 7/16inches) provided with the kit. Using too much force may damage thescrews. See Figure 44 to locate the screws, A and B.

Figure 44. Arrows A and B point to the two captive screws on the fluorescencemodule.

6. Insert one of the Fluorescence Optical Kits into the opening found on thedetection head. Push in completely.

7. Close the instrument door.

8. Power on the instrument.

9. To ensure the instrument detects the newly installed module, verify theFluorescence icon appears on the Instrument Status bar on theInstrument Control screen. See Figure 45.

Note: To prevent dust from accumulating on the optics, always keep anOptical Kit in the Fluorescence Module and keep the instrument doorclosed.

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B


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Figure 45. Detection Mode icon on the Instrument Status bar indicating thefluorescence module is detected.

17.C. Absorbance Module

The Absorbance Module is required for absorbance detection. If yourinstrument does not have this module, contact Promega Technical Services forinformation about purchasing and installation of the Absorbance Module. SeeSection 22.D for ordering information.

Installing the Absorbance Housing Unit

1. Touch the Door button to automatically close the instrument door andreset the Microplate Sample Tray to Home position.

2. Power off the GloMax-Multi+ Detection System and disconnect the powersupply from the back of the instrument.

Figure 46. Removing the metal cover plate.

3. Gently tip the GloMax-Multi+ Detection System so that it rests on its backpanel so that the bottom of the instrument is accessible.

4. Using a 3/16 inch Allen wrench remove the four 3/16 inch Allen headscrews from the metal plate cover exposing the interior (Figure 46).

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5. Carefully insert the connector on the Visible Absorbance housing unit intothe connector inside the bottom of the GloMax-Multi+ Detection System(Figure 47).

Figure 47. Attaching the Visible Absorbance housing unit.

6. Using the provided 3/16 inch Allen wrench, tighten the four 3/4 inchAllen head screws.

7. Manually open the instrument door and install the Visible AbsorbanceModule.


9. To ensure that the instrument is detecting the newly installed VisibleAbsorbance Module, verify that the Absorbance icon appears on theInstrument Status bar of the Instrument Control screen.

Installing the Absorbance Module

1. If the door is open, touch the Door button to automatically close theinstrument door and set the Microplate Sample Tray into Home position.

2. Turn off the GloMax-Multi+ Detection System.

3. Manually open the instrument door, and hold it open.

4. Hold the Absorbance Module with the fingertips of one hand. Guide theAbsorbance Module into the threaded stainless steel post located on the leftside of the instrument interior. See Figure 48.

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Figure 48. Installing the Absorbance Module.

5. Slide the Absorbance Module into position. Be cautious to not damage theprinted circuit board on the back of the detection head.

6. Hand tighten the two captive screws using the Allen wrench (size 7/16 inches)provided with the kit. Too much force may damage the screws. SeeFigure 49.

Figure 49. Arrows point to two captive screws on the Absorbance Module.

7. Close the instrument door.


9. To ensure the instrument detects the newly installed module, verify theAbsorbance icon appears on the Instrument Status bar on the InstrumentControl screen. See Figure 50.

Figure 50. Detection Mode icon on the Instrument Status bar indicating theAbsorbance Module is detected.

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6. Using the provided 3/16 inch Allen wrench, tighten the four 3/4 inch Allenhead screws.

7. Using the provided 1/4 inch Allen wrench, remove all four feet from the

bottom of the GloMax-Multi+ Detection System (Figure 53).8. Reattach each of the Instrument feet and the provided spacer with the 1.5

inch Allen head screw using the provided 1/4 inch Allen wrench(Figure 54).

9. With all four extended instrument feet attached, set the GloMax-Multi+Detection System back on its feet.

10. Manually open the instrument door and install the UV-Vis AbsorbanceModule. See Section 17.C for complete instructions.


Figure 53. Removing the feet from the bottom of the instrument.

12. To ensure that the instrument is detecting the newly installed UV-VisAbsorbance Module, verify that the Absorbance icon appears on theInstrument status bar of the Instrument Control screen (Figure 50).

Figure 54. Spacer and 1.5 inch Allen head screw.

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Figure 55. Instrument foot and spacer attached to bottom of instrument.

18. Injector System Installation and Operation

18.A. Injector System Components

Unpack the GloMax-Multi+ Injector System carefully. Verify all componentsand accessories have been received. See the component list enclosed with theinjector system for details.

The components included are shown in Figure 56.

Figure 56. GloMax-Multi+ Detection Injector System and accessories.

7963TA

Injector System Console Inlet Tubing Assembly

Outlet Tubing Assembly Waste Collection Tray

DB-15 Data Cable

Inlet Tube Holder

VerticalSupport Rod


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18.B. Installation Pro