Molecular Devices ImageXpress Micro User Manual
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Molecular Devices ImageXpress Micro User Manual

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ImageXpress
Micro
®
High Content Imaging System
Options
User Guide
5014166 D
September 2015

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Summary of Contents for Molecular Devices ImageXpress Micro

  • Page 1 ImageXpress Micro ® High Content Imaging System Options User Guide 5014166 D September 2015...
  • Page 2 For research use only. Not for use in diagnostic procedures. The trademarks mentioned herein are the property of Molecular Devices, LLC or their respective owners. These trademarks may not be used in any type of promotion or advertising without the prior written permission of Molecular Devices, LLC.

  • Page 3: Table Of Contents

    Contents Foreword ..........3 Who this User Guide is For.
  • Page 4 ImageXpress Micro High Content Imaging System Options User Guide Chapter 3: Environment Control and Fluidics Options Maintenance ........45 Carbon Dioxide Tubing .

  • Page 5: Who This User Guide Is For

    ImageXpress Transmitted Light option The ImageXpress Environment Control option, ImageXpress Fluidics option, and ImageXpress Transmitted Light option described in this guide are available for only the ImageXpress Micro High Content Imaging System. Conventions In this user guide, the following typographical conventions are used.
  • Page 6 ImageXpress Micro High Content Imaging System Options User Guide MetaXpress Software Use the MetaXpress Software with the ImageXpress Micro System to ® select a standard image analysis routine or to develop a custom protocol to fit your specific acquisition and analysis needs. The MetaXpress Software workflow is divided into two major parts: acquisition and analysis.

  • Page 7: Obtaining Support

    +1 408-747-1700. In Europe call +44 (0) 118 944 8000. Part of effective communication with Molecular Devices is determining the channels of support for the ImageXpress Micro System, including the MetaXpress Software. Molecular Devices provides a wide range of support: •...
  • Page 8 ImageXpress Micro High Content Imaging System Options User Guide • Technical Support: Phone: Contact Technical Support at (800)-635-5577 (U.S. only) or +1 408-747-1700. In Europe call +44 (0) 118 944 8000. Online: Visit http://www.moleculardevices.com/support follow the links in the knowledge base to the Technical Support Request Form to send an email to a group of experienced Technical Support representatives.

  • Page 9: Operations

    Environment Control Option Operations The ImageXpress Environment Control option is designed to maintain an ® environment for living cells to enable multi-day live-cell timelapse imaging. Temperature, carbon dioxide, and humidity can all be maintained within the sample plate so that cells can be kept alive for many days, growing at a rate comparable to that expected in a standard cell culture incubator.

  • Page 10: Environment Control Hardware

    ImageXpress Micro High Content Imaging System Options User Guide Environment Control Hardware The ImageXpress Environment Control option consists of a sealing ring on top of the sample plate and a top door above the plate that together forms a small, sealed volume. Humidified carbon dioxide is sourced into this small volume to form the required environment above the plate.
  • Page 11 Environment Control Option Operations Figure 1-1: Environmental Control Option Controller used for the ImageXpress Micro Standard and XLS Systems Figure 1-2: Systems Power & Options Controller used for the ImageXpress Micro Confocal System Note: The Compound Plate Temperature setting is available only when the Fluidics option is also installed.
  • Page 12 Figure 1-3: Environment Control option warm air hose and carbon dioxide tubing on the back of the Environmental Control Option Controller Figure 1-4: Environment Control option warm air hose and carbon dioxide tubing on the back of the ImageXpress Micro Standard System 5014166 D...
  • Page 13 Environment Control Option Operations Figure 1-5: Environment Control option water reservoir Figure 1-6: Environment Control option live-cell sealing ring 5014166 D...

  • Page 14: Items To Be Provided By The Customer

    ImageXpress Micro High Content Imaging System Options User Guide Items to Be Provided by the Customer The customer must provide the following items for the Environment Control option installation: • A tank of pre-mixed 5% CO and 95% air. •...

  • Page 15: Setting Up The Water Reservoir

    After you release , the current temperature in the chamber is displayed. Note: For the ImageXpress Micro Confocal System, you can set the temperature with the Instrument power switch in the On position. To regulate the temperature in the chamber, the Sample Heater power switch must be also in the On position.
  • Page 16 ImageXpress Micro High Content Imaging System Options User Guide To set up the water reservoir 1. In the MetaXpress software, click Screening > Plate acquisition and Control > Eject Plate to open the top door of the instrument. 2. Remove the insert that surrounds the top door.

  • Page 17: Setting Up The Carbon Dioxide

    4. Turn on the CO regulator to approximately 15 PSI to 20 PSI. 5. From the top of the ImageXpress Micro System, lift the insert surrounding the top door and remove it. Keep the top door of the instrument closed.

  • Page 18: Loading The Sample Plate

    ImageXpress Micro High Content Imaging System Options User Guide Loading the Sample Plate You must use the live-cell sealing ring to maintain the CO flow to the sample plate. The sealing ring helps to contain the inputted air directly over the cells and maintains the proper CO and temperature levels.
  • Page 19 Environment Control Option Operations 3. Place the live-cell sealing ring directly on top of the plate, making sure that it fits securely onto the plate. Figure 1-9: Environment Control option live-cell sealing ring 4. In the MetaXpress software, click Screening > Plate acquisition and Control >...
  • Page 20 ImageXpress Micro High Content Imaging System Options User Guide Before you run an experiment: • Allow the instrument to warm up for at least two hours. ° • Make sure the plate sealing ring is at 37 C before use, either by keeping it inside the instrument or inside an incubator.

  • Page 21: Environment Control Software

    Environment Control Option Operations Environment Control Software The MetaXpress software controls the ImageXpress Micro System screening and monitors the system’s environmental parameters. You can check Environment Control settings and connections in the MetaXpress software. To check the Environment Control settings 1.

  • Page 22: Environmental Parameters In Image Information

    ImageXpress Micro High Content Imaging System Options User Guide Environmental Parameters in Image Information The MetaXpress Software records environmental information when images are captured as shown in Figure 1-11 on page Note: There are wire sensors in the system to detect water, but the software does not record this as an image annotation.

  • Page 23: Doing Timelapse Experiments

    Environment Control Option Operations Doing Timelapse Experiments The ImageXpress Environment Control option incubates live cells, enabling imaging experiments to be performed over hours or days. Wells can be imaged repeatedly at fixed time intervals, and movies can be constructed from a series of images. Note: In a timelapse experiment, the time starts when the first well is acquired.
  • Page 24 ImageXpress Micro High Content Imaging System Options User Guide 4. Under Configure, click Timelapse to configure timelapse acquisition options. See Figure 1-13 on page Figure 1-13: Timelapse tab 5. In the Number of time points field, specify the number of time points to use.
  • Page 25 Environment Control Option Operations 7. If applicable, select additional options on the Autofocus tab. See Figure 1-14 on page 25. These options include:  First timepoint only: Recommended for fast kinetic experiments.  All timepoints: Recommended for long timelapse experiments. ...
  • Page 26 ImageXpress Micro High Content Imaging System Options User Guide 8. If applicable, select additional options on the Wavelength tabs. Wavelength tabs provide options for acquiring individual wavelength images during a timelapse experiment. See Figure 1-15 on page These options include: ...

  • Page 27: Reviewing Timelapse Data

    Environment Control Option Operations Reviewing Timelapse Data As with other ImageXpress System data, timelapse data is displayed in the MetaXpress software Review Plate Data dialog. To review the timelapse data 1. In the MetaXpress software, click Screening > Review Plate Data. Figure 1-16: Review Plate Data dialog 2.
  • Page 28 ImageXpress Micro High Content Imaging System Options User Guide Note: To clear the selection, click Clear Selection at the bottom of the dialog. 6. Click Load Selected Images. In the image window, you can view each time point as a plane in the stack using the navigation buttons above the image pane.
  • Page 29 Environment Control Option Operations To make a movie 1. Before you begin, load timelapse images and create a stack. See Reviewing Timelapse Data on page 2. Select the image window of the stack on the MetaXpress Software desktop. 3. In the MetaXpress software, click Stack > Make Movie. In the Simplified Menu Structure, click Edit >...
  • Page 30 ImageXpress Micro High Content Imaging System Options User Guide 5014166 D...

  • Page 31: Fluidics Hardware

    Fluidics Option Operations The ImageXpress Fluidics option consists of a single-channel, fluidics ® robot. This robot picks up tips from a tip box and draws fluid from and delivers fluid to a given well on the compound and sample plates. For the sample plate, the solenoid-operated plate shutter opens briefly, allowing the robot access to a given well without compromising the environmental control.
  • Page 32 ImageXpress Micro High Content Imaging System Options User Guide • Tip stripper. Used for ejecting tips. • Waste bin. Used to contain ejected tips and discarded liquid. • Doors with safety interlocks. Prevents doors from being opened during operation. Figure 2-1: Fluidics hardware...

  • Page 33: Items To Be Provided By The Customer

    Plates The tip and compound plate region inside the environmental enclosure can accommodate up to two compound plates. Molecular Devices recommends using standard 96-well or 384-well plates. U-bottom and V- bottom plates are beneficial when withdrawing and dispensing very small volumes of compound.
  • Page 34 4. In the right pane of the User Settings for hardware configuration dialog, click ImageXpress Micro Fluidics, and then click Settings. Figure 2-2: ImageXpress Micro Fluidics Settings dialog 5. In the ImageXpress Micro Fluidics Settings dialog, If applicable, change the tip adapter or the COM port. 5014166 D...

  • Page 35: Configuring Fluidics Software

    Fluidics Option Operations Configuring Fluidics Software (Stations and Properties) To use the ImageXpress Fluidics option, you must configure fluidics ® stations and properties. Note: The Configure Fluidic Stations functions are available at Devices > Configure Fluidic Stations, in the simplified menu, Control > ImageXpress >...
  • Page 36 ImageXpress Micro High Content Imaging System Options User Guide 2. On the Plate Acquisition Setup dialog, on the Configure tab, open the Acquisition tab. Figure 2-4: Plate Acquisition Setup dialog: Configure tab, Acquisition tab 3. Under Acquisition, select Use Fluidics.
  • Page 37 Fluidics Option Operations 5. Click Configure Stations. Figure 2-6: Configure Fluidics Stations dialog 6. On the Configure Fluidics Stations dialog, for each Tip Tray, select a Tip Type. Note: The system comes pre-configured with settings for FLIPR Tetra 96 and 384 tips. 7.
  • Page 38 ImageXpress Micro High Content Imaging System Options User Guide 8. If applicable, for the Sample Plate, type a value for X, Y, and Z offsets from the well center (in μm) for the sample plate. Type a positive Z offset to have the pipettor sample higher than normal.
  • Page 39 Fluidics Option Operations 12. On the Fluidics System Properties dialog, leave the default values for the fluidic system properties set as-is, or modify any values as needed. Table 2-1: Fluidic Systems Property values Option Description Draw Rate (μl/s) The rate at which the pump draws fluid. Dispense Rate (μl/s) The rate at which the pump dispenses fluid.

  • Page 40: Manual Fluidics Control

    ImageXpress Micro High Content Imaging System Options User Guide 13. Click OK to close the Fluidics System Properties dialog and return to the Configure Fluidics Stations dialog. 14. If applicable, click Define Tips and in the Define Tips dialog, define additional tip types, and then click Save.
  • Page 41 Fluidics Option Operations To control fluidics manually 1. In the MetaXpress software, click Devices > Fluidic Control. In the simplified menu, click Control > ImageXpress > Fluidic Control. 2. In the Fluidic Control dialog, specify the action you would like to do.
  • Page 42 ImageXpress Micro High Content Imaging System Options User Guide To record the steps in a journal 1. In the MetaXpress software, on the Journal menu, click Start recording. 2. Do the steps that you want to record. See To control fluidics manually on page 3.

  • Page 43: Doing Fluidics Experiments

    Fluidics Option Operations Doing Fluidics Experiments Fluidics experiments are defined by adding events to the Scheduled Events list. You can add three types of events: Compound Addition, Washout, and Journal. Fluidics events can be performed on all wells being imaged, or on a subset of wells. For example, you can add different amounts of compound to different wells, or treat wells for different amounts of time.
  • Page 44 ImageXpress Micro High Content Imaging System Options User Guide Figure 2-12: Adding events to a fluidics experiment 4. Click Add new Event to add an event to the Scheduled Events field. 5. In the Fluidic Event dialog, specify the time point at which you want the event to occur, and then specify whether the event should occur before or after the time point.
  • Page 45 Fluidics Option Operations Figure 2-13: Time point setting in Fluidic Event dialog For example, if you select time point 5 and select Before imaging, then the fluidic event occurs before the time point. If you select After imaging, then the fluidic event occurs after the time point.
  • Page 46 ImageXpress Micro High Content Imaging System Options User Guide 6. In the Fluidic Event dialog, select the event type and its properties. Event types include:  Compound addition: A Compound addition event, shown in Figure 2-15 on page 46, draws liquid from the compound plate and adds it to the sample plate.
  • Page 47 Fluidics Option Operations  Washout: A Washout event removes liquid from the sample plate and replaces it with liquid from the compound plate. There is a one-to-one mapping between the wells as shown Figure 2-16. An exchange consists of a washout event in which fluid is removed from the sample plate and discarded, and fresh media or fluid is placed in the well.
  • Page 48 ImageXpress Micro High Content Imaging System Options User Guide  Journal: A Journal event is used for any special cases or custom protocols (for example, experiments not using a one- to-one mapping of wells, or drawing once and dispensing multiple times) as shown in Figure 2-17.

  • Page 49: Preventing Evaporation From Compound Plates

    Evaluated Seals Molecular Devices has evaluated both heat-applied and pressure-applied seals with a variety of polypropylene and polystyrene plates. Two examples are listed here. Other manufacturers might provide similar performance.
  • Page 50 ImageXpress Micro High Content Imaging System Options User Guide Pressure-applied seals G&L Precision Die Cutting pressure-applied aluminum foil seals • Part number: G&L Aluminum Microplate Liddings (GL-255) • For specifications, ordering information, pricing, and sealing instructions, contact G&L Precision (www.glprecision.com) •...

  • Page 51: Chapter 3: Environment Control And Fluidics Options Maintenance

    For additional information about cleaning the system, see the ImageXpress Micro Widefield High Content Imaging System User Guide or the ImageXpress Micro Confocal High Content Imaging System User Guide.

  • Page 52: Carbon Dioxide Tubing

    ImageXpress Micro High Content Imaging System Options User Guide Carbon Dioxide Tubing The tubing within the environmental enclosure should be cleaned only if moisture has collected in the tubing or visual inspection of the tubing suggests contamination. There are two tubing sections associated with carbon dioxide delivery.

  • Page 53: Cleaning The Water Reservoir

    Environment Control and Fluidics Options Maintenance Cleaning the Water Reservoir Air supplied to the environmental enclosure flows through the water reservoir, ensuring the air has a high degree of humidity. To clean the water reservoir Required tool: Phillips screwdriver 1. In the MetaXpress software, click Screening > Plate acquisition and Control >...

  • Page 54: Waste Disposal Box And Plate Sealing Ring

    ImageXpress Micro High Content Imaging System Options User Guide Waste Disposal Box and Plate Sealing Ring Proper cleaning of the waste disposal box and the plate sealing ring is highly dependent on the type of waste being disposed of and whether the box is being used with a protective liner.

  • Page 55: Chapter 4: Transmitted Light Option Operations

    Transmitted Light Option Operations The ImageXpress Transmitted Light option is designed for phase- ® contrast transmitted-light imaging. In general terms, transmitted light is used to describe microscopy in which light is transmitted from a source on one side of the specimen, with the objective on the other side. Typically the light is first passed through a condenser so high illumination is focused on the specimen.

  • Page 56: Transmitted Light Hardware

    ImageXpress Micro High Content Imaging System Options User Guide Transmitted Light Hardware The ImageXpress Transmitted Light option consists of the following hardware components. • Nikon phase optics. Allows either phase contrast or brightfield imaging. See Figure 4-1 on page 57 Figure 4-2 on page •...
  • Page 57 Transmitted Light Option Operations Figure 4-1: ImageXpress Micro Standard System with the Transmitted Light option Figure 4-2: ImageXpress Micro Standard System with Transmitted Light option (detailed view) 5014166 D...
  • Page 58 ImageXpress Micro High Content Imaging System Options User Guide Figure 4-3: Transmitted Light shutter Figure 4-4: Hinge-release knob and hinge Item Description Hinge-release knob Hinge Hinge-release knob Assembly tipped back with hinge-release knob unscrewed 5014166 D...

  • Page 59: Transmitted Light Software Configuration

    Transmitted Light Option Operations Transmitted Light Software Configuration You can create illumination settings to turn the lamp on and off. Do the following procedure to create illumination settings named Transmitted Lamp Off and Transmitted Light, and to set a level for the lamp illumination.
  • Page 60 ImageXpress Micro High Content Imaging System Options User Guide 2. In the Configure Illumination dialog, create an illumination setting named Transmitted Lamp Off, select Lamp, ImageXpress Micro Transmitted Light, and set the lamp illumination to zero, as shown in Figure 4-6.

  • Page 61: Transmitted Light Phase Ring Alignment

    Software. See To use the phase ring adjustment lens on the ImageXpress Micro Confocal System on page To use the flip-in lens on the ImageXpress Micro Standard and XL Systems CAUTION! Wear gloves, and do not touch, move, or otherwise damage the mirror to the right of the flip-in lens.
  • Page 62 4. Move the flip-in lens into place for the alignment procedure. The flip-in lens must be out of the way for any actual imaging. Figure 4-8: Flip-in lens on the ImageXpress Micro Standard and XLS Systems 5. Power on the instrument.
  • Page 63 Transmitted Light Option Operations To use the flip-in lens on the ImageXpress Micro XLS System 1. Power off the instrument. 2. Use a Phillips screwdriver to remove both of the left side panels of the instrument to access the flip-in lens mounted on the outside of the optics cover.
  • Page 64 ImageXpress Micro High Content Imaging System Options User Guide Figure 4-11: Flip-in lens extended for phase ring alignment 7. Slide the flip-in lens assembly onto the optics cover with the flip-in lens on the inside for the phase ring alignment.
  • Page 65 Transmitted Light Option Operations To use the phase ring adjustment lens on the ImageXpress Micro Confocal System 1. Exit the MetaXpress Software if it is running. 2. Click Start > Programs > MetaXpress and then right-click Meta Imaging Series Administrator and select Run as administrator to start the MetaXpress Meta Imaging Series Administrator software.
  • Page 66 ImageXpress Micro High Content Imaging System Options User Guide 6. In the IXConfocal Module Emission Wheel Settings dialog, set the Filter Exchange Mode to Off. Figure 4-13: The IXConfocal Module Emission Wheel Settings dialog 7. Click Open Control Dialog. Figure 4-14: IXConfocal Module Emission Wheel Control dialog 8.
  • Page 67 Transmitted Light Option Operations To align the phase ring Note: For the ImageXpress Micro Confocal System, make sure that the instrument is in widefield mode before aligning the phase ring. 1. Make sure that the phase ring adjustment lens is in place.
  • Page 68 ImageXpress Micro High Content Imaging System Options User Guide 3. Turn the adjustment ring, which is located below the arm of the Transmitted Light tower, to the middle of the aperture range. Figure 4-16: Aperture stop position for phase ring alignment...
  • Page 69 Transmitted Light Option Operations 4. Move the field-stop slider, located just above the A label on the condenser turret, all the way to the right. Figure 4-17: Field stop slider 5. Select the diffusion filter, or place some lens paper or lint-free wipes on top of the condenser lens to serve as a diffuser.
  • Page 70 ImageXpress Micro High Content Imaging System Options User Guide Figure 4-18: Out-of-focus image of phase contrast ring 11. Adjust the position of the Z-stage to obtain a reasonably sharp image of the objective phase ring. Figure 4-19: In-focus image of phase contrast ring...
  • Page 71 Transmitted Light Option Operations 12. Use the region tools in the MetaXpress software to mark the position of the phase ring. Make sure that you capture both horizontal and vertical axes. Use either the ellipse region tool or multiple line tools. See Figure 4-20.
  • Page 72 ImageXpress Micro High Content Imaging System Options User Guide 15. Snap an image. The condenser annulus ring shows. See Figure 4-21. You might need to adjust the exposure time. Figure 4-21: Image of condenser annulus ring 16. In the MetaXpress software, click Live Mode in either the Acquire or Plate Acquisition and Control dialog.
  • Page 73 If required, adjust the lamp power or exposure level. 21. In the MetaXpress software, click Stop Live. 22. For the ImageXpress Micro Standard, XL, or XLS Systems, move the flip-in lens out of the optical path. ...

  • Page 74: Transmitted Light Experiments

    ImageXpress Micro High Content Imaging System Options User Guide Transmitted Light Experiments Do the following procedure if you are running an experiment with Transmitted Light as one of the selected wavelengths. Transmitted light acquisition is compatible with the confocal modes in the ImageXpress Micro Confocal System.

  • Page 75: Index

    Index doors with safety interlocks 32 draw overfill 40 draw rate 40 aperture stop 68 environmental control carbon dioxide hardware 10 environmental control 10 environmental control option 9 recording 21 evaporation prevention 49 setting up 17 experiments 43 tubing 52 cleaning tubing 52 components, customer-provided...
  • Page 76 ImageXpress Micro High Content Imaging System Options User Guide humidity environmental control 10 phase contrast objectives 57 transmitted light 55 phase ring alignment 61 illumination settings 59 plate heaters 31 interlocks 32 plate region 33 plate sealing ring 54 plates...
  • Page 77 Index shutter 31 simplified menu structure 6 single-channel pipetting 31 washout event 47 systems power and options waste bin 32 controller waste disposal box 54 environmental control 11 water reservoir setting up 15 wet dispense 40 white light lamp 56 temperature recording 22 setting up 14...
  • Page 78 The trademarks used herein are the property of Molecular Devices, LLC or their respective owners. ©2015 Molecular Devices, LLC. Specifications subject to change without notice. Patents: www.moleculardevices.com/productpatents All rights reserved. FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES...

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