Page 1 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 5084773 A March 2021...
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
Water Immersion Option Safety Chapter 1: ImageXpress Confocal HT.ai System ImageXpress Confocal HT.ai System Features MetaXpress Software Features Theory of Operation Chapter 2: Using the ImageXpress Confocal HT.ai System Starting the System Understanding Status Lights Maintaining the Instrument Shutting Down the System...
Page 4 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Plate Acquisition Setup Dialog: Configure Tab, Objective and Camera Tab Plate Acquisition Setup Dialog: Configure Tab, Plate Tab Plate Acquisition Setup Dialog: Configure Tab, Sites to Visit Tab Plate Acquisition Setup Dialog: Configure Tab, Acquisition Tab...
Page 5 Appendix A: Instrument Specifications Appendix B: Compatible Objectives Appendix C: Filter Specifications and Calibration Compatibility Appendix D: Optional Parts and Accessories Appendix E: File Privileges for the MetaXpress Software Appendix F: Robotic Plate Handling Verifying External Control Settings Appendix G: Electromagnetic Compatibility 5084773 A...
Page 6 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 5084773 A...
Page 7: Safety Information
Always remember that the key to safety is operating the instrument with care. DANGER! If the instrument is used in a manner not specified by Molecular Devices, the protection provided by the equipment might be impaired. Safety Symbols in This Guide All safety symbols in the user guide are framed within a triangle.
Page 8: Safety Labels On The Instrument
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Safety Labels on the Instrument Each safety label found on the instrument contains an alert symbol that indicates the type of potential safety hazard related to the label. The following table lists the alert symbols that can be found on Molecular Devices instruments.
Page 9: Protective Housing And Safety Interlocks
Safety Interlock Failure If the focusing laser stays on when the top door of the instrument is open, it is unsafe to continue using the instrument due to a safety interlock failure. Contact Molecular Devices Support immediately. See Obtaining Support on page 237.
Page 10: Laser Safety
Never attempt to open the instrument or access the high-power laser products. Installation and service is to be performed by a Molecular Devices Field Service Engineer only. The Class 4 laser product is designed and tested in accordance with IEC 61010-1, CAN/CSA C22.2 Number 61010-1, CENELEC EN 61326-1:2006, and FCC Part 15 Subpart B, Class A.
Page 11 Safety Information The ImageXpress Confocal HT.ai system is equipped with a redundant laser safety system. When samples are being loaded or unloaded, hardware interlocks prevent the laser modules from powering on until the instrument top door is closed. DANGER! VISIBLE AND/OR INVISIBLE LASER RADIATION.
Page 12: Electrical Safety
We recommend that you power off the instrument when it is not in use. WARNING! The ImageXpress Confocal HT.ai system is an Equipment Class 1 product that relies on protective earth grounding for safe operation. Any interruption of the protective earth ground conductor—inside or outside the instrument—or disconnection of the protective earth ground terminal can result in personal injury.
Page 13: Moving Parts Safety
CAUTION! Moving the instrument can disrupt sensitive optical alignments. We recommend that you contact Molecular Devices Technical Support to schedule a Field Service Engineer to help with moving your instrument. Your warranty or service contract does not cover problems caused during or as a result of shipment or relocation of the instrument.
Page 14: Chemical And Biological Safety
Review the Chemical and Biological Safety information contained in this guide. See Chemical and Biological Safety on page Perform only the maintenance tasks described in this guide. Contact Molecular Devices to inspect and perform preventive maintenance service on the instrument each year. See Obtaining Support on page 237.
Page 15: Water Immersion Option Safety
CAUTION! Do not remove or change the water immersion objectives that are installed within the instrument. A Molecular Devices Field Service Engineer must install and remove water immersion objectives. Do not touch the leak detection sensor, which is the brass color ring around the top of the objective below the cap.
Page 16 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 5084773 A...
Page 17: Chapter 1: Imagexpress Confocal Ht.ai System
Chapter 1: ImageXpress Confocal HT.ai System The ImageXpress® Confocal HT.ai High-Content Imaging System is an integrated cellular imaging and analysis system designed for rapid, automated screening of fluorescently labeled biological samples in microplates. With the addition of modular options, the system provides environmental control for live cell imaging and uses transmitted light capability for label-free imaging.
Page 18: Imagexpress Confocal Ht.ai System Features
ImageXpress Confocal HT.ai System Features Figure 1-1: ImageXpress Confocal HT.ai System Components without Options Illumination System: Excitation The ImageXpress Confocal HT.ai system is equipped with an external light source connected to the instrument with a light guide. External Light Source The external light source is a Class 4 high-power laser product with a rated lifetime of 20,000 hours.
Page 19 A linear encoder that features better than 100 nm resolution monitors the X-Y stage position. Autofocus Laser The ImageXpress Confocal HT.ai system uses a laser to autofocus the image. The red (690 nm) diode laser projects a laser spot onto the sample. Reflections of this spot from the bottom of the plate and the plate-sample interface are imaged by a dedicated, fast-focus sensor and the spot acts as a reference for focus software.
Page 20 The emission wavelength range is 400 nm to 750 nm. Camera The ImageXpress Confocal HT.ai system uses a scientific CMOS camera with a 2048 × 2048 image sensor format (6.5 × 6.5 µm pixel size) and a peak quantum efficiency of 82%. Confocal Disks Three Confocal disk box configurations are available for the ImageXpress Confocal HT.ai...
Page 21: Metaxpress Software Features
Chapter 1: ImageXpress Confocal HT.ai System MetaXpress Software Features Use the MetaXpress software with the ImageXpress Confocal HT.ai system to select standard acquisition and analysis protocols or to develop custom protocols to fit your acquisition and analysis needs. The MetaXpress software workflow has two major parts: acquisition and analysis.
Page 22: Theory Of Operation
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Theory of Operation Figure 1-2: ImageXpress Confocal HT.ai System Optical Path The ImageXpress Confocal HT.ai system uses the following components and functions: Fluorescence Imaging, see page 23 Confocal Imaging, see page 23...
Page 23 Excitation and Emission Filters In the ImageXpress Confocal HT.ai system, the excitation filters are located inside the light source. The emission filters and the dichroic mirrors are located within two separate, controllable wheels inside the confocal module.
Page 24 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Dichroic Mirror In the ImageXpress Confocal HT.ai system, framed dichroic mirrors are in a five-position filter wheel. See Figure 1-2. A dichroic mirror is a specially designed beam splitter that transmits light above a certain cutoff wavelength and reflects light at shorter wavelengths.
Page 25 The ImageXpress Confocal HT.ai system can be configured with high-quality Nikon objectives. Compatible Objectives on page 243. If the objective you want to use is not listed, contact Molecular Devices to verify compatibility with your system. Note: Some objectives have adjustable spherical aberration correction collars for imaging through thick substrates such as most microplates.
Page 26 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 5084773 A...
Page 27: Chapter 2: Using The Imagexpress Confocal Ht.ai System
Chapter 2: Using the ImageXpress Confocal HT.ai System This section provides an overview of the workflow for using the ImageXpress Confocal HT.ai system. Starting the System, see below Understanding Status Lights, see page 33 Maintaining the Instrument, see page 34...
Page 28 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Powering On the External Light Source Power on the external light source before you start the software. DANGER! The external light source is a Class 4 high-power laser product. See Laser Safety on page 10 for details on the Class 4 high-power laser product and specific warning and danger statements for working with the external laser light source.
Page 29 Chapter 2: Using the ImageXpress Confocal HT.ai System To power on the external light source: 1. Confirm that the MetaXpress software on the acquisition computer is shut down. 2. Confirm that the instrument is running. 3. Confirm that the following instrument interlocks are closed: All instrument doors are closed.
Page 30 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 7. Press the POWER/STATUS button (5). The status indicator briefly turns yellow, then red. 8. Turn the master key switch to the ON position. The status indicator briefly turns yellow, then green. 9. Wait about 30 seconds until both LASER READY indicators (4) turn blue.
Page 31 Chapter 2: Using the ImageXpress Confocal HT.ai System Starting the Software Finally, start the MetaXpress software. To start the software: 1. Do one of the following: a. Double-click the MetaXpress icon on the computer desktop. b. Click Start > MetaXpress 6 > MetaXpress.
Page 32 Check that the ImageXpress Systems Power and Options Controller and the external light source are powered on, if present. Restart the system. If the message continues, use AxoTrace and contact Molecular Devices Technical Support. Logging AxoTrace Software Messages to .txt Files on page 239.
Page 33: Understanding Status Lights
Chapter 2: Using the ImageXpress Confocal HT.ai System Understanding Status Lights The status light on the ImageXpress Confocal HT.ai instrument provides information on the instrument status. You can turn off the status lights. See Turning Off the Status Indicator Lights on page 236.
Page 34: Maintaining The Instrument
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Maintaining the Instrument User maintenance tasks for the ImageXpress Confocal HT.ai system include cleaning the instrument, changing filters, and changing and cleaning objectives. Perform only the maintenance tasks described in this guide. Contact Molecular Devices Technical Support to inspect and perform preventive maintenance service on the instrument each year.
Page 35: Chapter 3: System Installation And Testing
Chapter 3: System Installation and Testing The ImageXpress Confocal HT.ai system ships fully configured. A Molecular Devices Field Service Engineer installs the system at your site. The base system includes the instrument, laser light source, acquisition computer, and accessory kit. See...
Page 36: Verifying Device Settings In The Meta Imaging Series Administrator Software
Verifying Device Settings in the Meta Imaging Series Administrator Software Use the Meta Imaging Series Administrator Software to configure all hardware and software components. This section describes how to verify that the ImageXpress Confocal HT.ai system hardware components are properly configured in the Meta Imaging Series Administrator Software and in the MetaXpress software.
Page 37 Chapter 3: System Installation and Testing 3. In the List of Groups list, select MetaXpress. Figure 3-2: Meta Imaging Series Administrator Software Home Dialog 4. Click Configure Hardware to display the Configure Hardware dialog. 5. Click Configure Devices to display the User Settings dialog. Figure 3-3: User Settings Dialog 5084773 A...
Page 38 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 6. In the Claimed Devices list, select ImageXpress Micro X and then click Settings to display the ImageXpress Micro X Settings dialog. Figure 3-4: ImageXpress Micro X Settings Dialog 7. In the Move field, increase the step size to 10,000 µm.
Page 39 Chapter 3: System Installation and Testing 11. In the Claimed Devices list, select ImageXpress Micro Y and then click Settings to display the ImageXpress Micro Y settings dialog. Figure 3-6: ImageXpress Micro Y Settings Dialog 12. In the Move field, increase the step size to 10,000 µm. 13.
Page 40 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 16. In the Claimed Devices list, select ImageXpress Micro Z and then click Settings to display the ImageXpress Micro Z setting dialog. Figure 3-8: ImageXpress Micro Z Settings Dialog 17. Verify that the value in the Device Units field is 50.
Page 41: Verifying Camera Settings In The Meta Imaging Series Administrator Software
32. Click OK to display the Configure Hardware dialog and proceed to the next section. Verifying Camera Settings in the Meta Imaging Series Administrator Software To verify the installation of the correct version of the ImageXpress Confocal HT.ai system camera driver: 1.
Page 42: Verifying And Backing Up Settings In The Metaxpress Software
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Verifying and Backing Up Settings in the MetaXpress Software After you confirm hardware settings in the Meta Imaging Series Administrator Software, check the settings in the MetaXpress software. See the following sections:...
Page 43 Chapter 3: System Installation and Testing Verifying Illumination Settings Use the Configure Illumination dialog to confirm illumination settings for the ImageXpress Confocal HT.ai system objectives before you use the system. To verify the illumination settings: 1. In the MetaXpress software, do one of the following to display the Configure Illumination dialog: From the simplified menu, click Control >...
Page 44 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Verifying Calibration Settings Use the Calibrate Distances dialog to verify calibration distances. To verify the calibration settings: 1. In the MetaXpress software, do one of the following to display the Calibrate Distances dialog: From the simplified menu, click Measure >...
Page 45 3. Make sure the appropriate magnification setting displays for each calibration. Note: Use the same calibration setting for both X and Y. You can use the following estimated values for ImageXpress Confocal HT.ai system calibration settings: Note: To measure pixel sizes more accurately, in the IX Taskbar, click System Maintenance and then click Measure Pixel Sizes.
Page 46 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Verifying the Laser Autofocus Sensor This procedure uses a bead plate to verify that the laser autofocus sensor responds. To verify that the Laser Autofocus sensor responds: 1. In the MetaXpress software, do one of the following to display the Plate Acquisition Setup dialog: From the simplified menu, click Screening >...
Page 47 Chapter 3: System Installation and Testing 15. In the Preview Pass image window, ensure that there is at least one peak. If the Preview Pass image window shows at least one peak, the Laser Auto Focus Sensor is enabled and functional. If the Preview Pass image window does not show any peaks, ensure that the plate is properly seated.
Page 48 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Verifying the Plate Reference Point (A1 Center) This procedure requires the metal slide holder plate that ships with the instrument to verify that the plate reference point is in the center of well A1.
Page 49 Figure 3-17: A1 Pinhole in Focus at 10X If the A1 pinhole is not centered, or if you cannot find the hole, contact Molecular Devices Technical Support. See Obtaining Support on page 237.
Page 50 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Verifying Plate Types Use this procedure to verify that the plate type files included with the software are available from the Plate Acquisition Setup dialog. To verify the plate types were installed: 1.
Page 51 Chapter 3: System Installation and Testing Confirming Laser Autofocus Settings for Plate Files Before you use a plate file, confirm that the laser autofocus settings are optimized for the plate. Note: If you use an objective with a correction collar, ensure that the correction collar is set appropriately for the plate.
Page 52 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Verifying Shading Correction Files-Legacy Note: This procedure is required if you select the Legacy Correction check box for one or more wavelengths in an acquisition protocol. If you select Legacy Correction as the Shading Correction method for an acquisition wavelength, you need the shading correction image files for each magnification and illumination setting.
Page 53: Chapter 4: Preparing For Acquisition
Chapter 4: Preparing for Acquisition This section provides general guidelines to consider before you acquire plate data. These guidelines help ensure that the images you acquire are the best possible quality. Consider the following criteria to obtain the best possible fluorescence image quality: Assay Design, see below Plate Selection, see page 55 Sample Preparation, see page 57...
Page 54 Uniformly staining thick samples for confocal imaging can be difficult. Organism-Based Assays You can use the ImageXpress Confocal HT.ai system to image whole organisms, such as nematodes or zebrafish. Treat the organisms with anesthetic, use a gel preparation, or both to immobilize the organisms for best results.
Page 55: Plate Selection
Plate Material WARNING! The ImageXpress Confocal HT.ai instrument uses a Class 4 high-power laser light source, which can cause materials to smolder or burn, especially at close range. Dark materials, which absorb heat, and materials such as plastic, paper, and fabric can easily be burned by visible laser beams.
Page 56 If one or more plate types do not work with your robot grippers, contact Molecular Devices for assistance. Also make sure that there is a consistent location to affix a barcode on the side of the plate.
Page 57: Sample Preparation
Chapter 4: Preparing for Acquisition Sample Preparation There are many variables involved in sample preparation. It is best to test these variables during the assay optimization phase before you prepare multiple plates for screening. This section describes considerations for imaging assays. Cell Density Cell density can affect the performance of the cells as well as downstream image analysis.
Page 58: Plate Acquisition Settings
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Plate Acquisition Settings You make most plate acquisition settings in the Plate Acquisition Setup dialog in the MetaXpress software. The settings are dependent on the content and distribution of the samples, as well as the requirements of the experiment.
Page 59 Shading is an artifact that can come from the objective, optics, light source, or background light from the room. The ImageXpress Confocal HT.ai system is designed to minimize shading effects and provides a shading correction feature within the software. This is recommended for assays where comparison of individual cell intensities is critical, such as a cell cycle assay.
Page 60 The ImageXpress Confocal HT.ai system uses a standard, selectable light source filter to create the correct excitation wavelength, and a filter wheel to select the corresponding emission wavelength filtration for the experiment.
Page 61: Instrument Maintenance
189. The basic design of the ImageXpress Confocal HT.ai system light source and light path within the instrument help ensure that the light that reaches the sample is the best possible quality. If you find that the light quality has become degraded, contact your system administrator or your Molecular Devices representative to correct the problem.
Page 62 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 5084773 A...
Page 63: Chapter 5: Configuring Plate Acquisition Options
MetaMorph® Microscopy Automation and Image Analysis Software. The MetaMorph Software contains numerous dialogs for image acquisition, processing, and acquisition. The MetaXpress software adds database integration and tools to control the ImageXpress Confocal HT.ai system and to acquire and analyze microplates.
Page 64: Accessing The Plate Acquisition Setup Dialog
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Accessing the Plate Acquisition Setup Dialog The MetaXpress software displays menus along the top of the screen. The Screening menu provides access to the dialogs that allow you to configure protocols and acquire images. The Screening menu also provides access to plate configuration dialogs including the Plate Acquisition Setup dialog.
Page 65: Plate Acquisition Setup Dialog Layout
The Plate Acquisition Setup dialog provides three primary functions: Use the Plate Map and Site Map area to view the plate layout and control some of the movable physical components of the ImageXpress Confocal HT.ai system. Use the Configure tab to configure the plate acquisition protocol.
Page 66 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Plate Map and Site Map The Plate Map area in the upper left is a graphical representation of the plate type you select. The following functionality is available above the plate map: Use the Protocol drop-down to select from the eight most recently used plate acquisition protocols.
Page 67 Use the Site Map to control some of the movable physical components of the ImageXpress Confocal HT.ai system and to configure the sites to acquire. Sites you select to acquire display green and sites you choose to not acquire display gray.
Page 68 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Configure Tab Use the Configure tab to configure the plate acquisition protocol. The Configure tab is organized in a “top-to-bottom” tab structure that runs down the left side to guide you through the process of setting up the protocol configuration.
Page 69: Plate Acquisition Setup Dialog: Configure Tab, Objective And Camera Tab
Chapter 5: Configuring Plate Acquisition Options Plate Acquisition Setup Dialog: Configure Tab, Objective and Camera Tab Use the Objective and Camera tab to set the objective magnification, the camera binning, and the acquisition mode for a protocol. Use these settings to improve either the image acquisition speed or the image quality.
Page 70 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Table 5-1: Configure Tab, Objective and Camera Tab Configuration Options Option Description Magnification Selects the magnification setting to assign X and Y offset values (parcentricity) and a Z offset (parfocality) to a specific objective. When you select an objective, the objective physically moves to the acquisition position.
Page 71 Chapter 5: Configuring Plate Acquisition Options Configuring Objective and Camera Options Tip: As you configure an acquisition protocol, you can open the Summary panel to view the current values for the protocol settings. See Viewing the Summary Panel on page 123.
Page 72: Plate Acquisition Setup Dialog: Configure Tab, Plate Tab
Use the Plate tab to configure the plate dimensions to accurately control the X, Y, and Z movements of the ImageXpress Confocal HT.ai system. Define accurate plate dimensions to prevent the instrument from making potentially hazardous movements. The plate dimensions you define also ensure that laser-based autofocus is as accurate as possible.
Page 73 Chapter 5: Configuring Plate Acquisition Options Table 5-2: Plate Settings Option Description Plate Name Specifies the plate type for the protocol. You can select a configuration or you can create a new plate configuration. The fields display values for the plate type you select.
Page 74 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Table 5-2: Plate Settings (continued) Option Description Plate Width Specifies the Plate width in mm. The ANSI standard is 85.5 mm. Row Offset Specifies the distance in µm between the center of well A01 and the top edge of the plate.
Page 75 Note: The Laser Autofocus Wizard calculates measurements as accurately as possible. Some manual verification and adjustment of the settings might be necessary to optimize the results, particularly for thin-bottom plates. See the application help or contact Molecular Devices Technical Support. See Obtaining Support on page 237.
Page 76 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Configuring the Dimensions for New Plates Use the Plate tab to define the dimensions for a new custom plate. If the plate is 96-well or 384-well format, select a similar plate template, and modify the values as needed. If the plate is any other size, you must specify all values for the plate.
Page 77: Plate Acquisition Setup Dialog: Configure Tab, Sites To Visit Tab
Chapter 5: Configuring Plate Acquisition Options Plate Acquisition Setup Dialog: Configure Tab, Sites to Visit Tab Use the Sites to Visit tab to configure the number of sites to acquire for each well in a plate acquisition protocol. You can acquire a single site per well or multiple sites per well. The number of sites that you can acquire in a well depends on the size of the well, the objective magnification, the distribution of sample material in the well, the type of plate, and the fluid content of the well.
Page 78 ImageXpress Confocal HT.ai High-Content Imaging System User Guide To acquire a single site in each well: 1. Select the Sites to Visit tab. 2. Select the Single Site Site Option. 3. Select the Custom Field of View (%) check box to specify a percentage of the field of view for the camera to acquire, and then enter the X (width) value and Y (height) value.
Page 79 Chapter 5: Configuring Plate Acquisition Options Acquiring a Fixed Number of Sites in Each Well Use the Sites to Visit tab to acquire a fixed number of sites in each well. Figure 5-9: Configure Tab, Sites to Visit Tab, Fixed Number of Sites Option To acquire a fixed number of sites in each well: 1.
Page 80 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 4. In the Columns field and the Rows field, enter the maximum number of sites to visit. For example, Columns = 2 and Rows = 2 acquires up to four sites, Columns = 3 and Rows = 3 acquires up to nine sites.
Page 81 Chapter 5: Configuring Plate Acquisition Options 7. To adjust spacing, do one of the following: To manually increase the spacing between adjacent sites, enter positive numbers in the Spacing fields or to overlap the sites, enter negative numbers in the Spacing fields.
Page 82 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Configuring Adaptive Acquisition for Well Sites The Sites to Visit tab Adaptive Acquisition option is a computational algorithm that counts the number of cells on the fly during sample acquisition. This increases the chances of collecting valid data in every well.
Page 83 Chapter 5: Configuring Plate Acquisition Options 5. Use the plate map to specify the wells for data acquisition. Active wells are green and inactive wells are gray. When the plate dimensions, plate skirt, and/or objective prevents acquisition from the wells at the edge of the plate, the affected wells are blocked, and display surrounded by a gray rectangle.
Page 84 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 7. To adjust spacing, do one of the following: To manually increase the spacing between adjacent sites, enter positive numbers in the Spacing fields or to overlap the sites, enter negative numbers in the Spacing fields.
Page 85 Chapter 5: Configuring Plate Acquisition Options 8. Specify the Adaptive Acquisition settings. Table 5-3: Adaptive Acquisition Settings Setting Description Minimum The minimum number of sites to visit in a well. The software acquires at least Sites to Visit this minimum number of sites and then it continues to acquire sites until it reaches the total number of cells counted per well.
Page 86 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 9. To determine if the acquisition settings are appropriate to differentiate nuclei, do the following: a. Select a well and site to review your settings. b. Click Focus to bring the sample into focus.
Page 87 Chapter 5: Configuring Plate Acquisition Options Calculating the Intensity Above Local Background Gray Levels The Sites to Visit tab Adaptive Acquisition option is a computational algorithm that analyzes the number of cells on the fly during sample acquisition. This increases the chances of collecting valid data in every well.
Page 88 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Configuring Multi-Well Acquisitions Run a multi-well acquisition to acquire high density plates faster. A multi-well acquisition uses the size of the camera field of view to acquire several wells simultaneously while taking a single image, thereby reducing plate acquisition time.
Page 89 Chapter 5: Configuring Plate Acquisition Options To run a multi-well acquisition: 1. Select the Sites to Visit tab. 2. Select the Multi-well Site Option. 3. Leave the Custom Field of View (%) check box clear. For multi-well acquisitions, always use the full field of view.
Page 90: Plate Acquisition Setup Dialog: Configure Tab, Acquisition Tab
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Plate Acquisition Setup Dialog: Configure Tab, Acquisition Tab Use the Acquisition tab to specify the autofocus options and additional acquisition options for the protocol. Most acquisition options you select add an additional tab to the Configure tab.
Page 91: Configuring Autofocus Options
Chapter 5: Configuring Plate Acquisition Options Configuring Autofocus Options The Acquisition tab provides two autofocus options: Laser-Based Focusing is generally set to find the bottom of the well, and then moves the objective a distance up from the well bottom. This method is the fastest and does not cause photo damage to your wells.
Page 92 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 4. Configure the Laser-Based Focusing options. Laser-Based Focusing is available when you select the Enable Laser-Based Focusing check box on the Acquisition tab. Table 5-4: Autofocus Tab, Laser-Based Focusing Options Option Description Configure Displays the Configure Laser Autofocus Settings dialog.
Page 93 Chapter 5: Configuring Plate Acquisition Options 5. Configure the Image-Based Focusing options. Image-Based Focusing is available when you select the Enable Image-Based Focusing check box on the Acquisition tab. Table 5-5: Autofocus Tab, Image-Based Focusing Options Option Description Algorithm Select one of the following focusing algorithm options: Standard: Based on a standard group of settings including a normal camera signal level.
Page 94 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 6. Configure the remaining options. Table 5-6: Autofocus Tab, General Options Option Description Initial Well for Sets the well for the first Find Sample autofocus. Select one of the following: Finding First Well Acquired: Finds the sample autofocus using the first well that is Sample acquired.
Page 95: Specifying The Number Of Acquisition Wavelengths
Chapter 5: Configuring Plate Acquisition Options Specifying the Number of Acquisition Wavelengths Use the Wavelengths tab to specify the total number of acquisition wavelengths for an experiment. You must specify at least one acquisition wavelength and you can specify up to eight.
Page 96: Configuring Acquisition Wavelengths
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Configuring Acquisition Wavelengths Use the W# tabs below the Wavelength tab to configure acquisition wavelengths. The sequence in which you configure the acquisition wavelengths corresponds to the acquisition order. For example, if you configure the W1 tab as a DAPI wavelength and then configure the W2 tab as a FITC wavelength, then the DAPI acquisition wavelength runs first and the FITC acquisition wavelength runs second.
Page 97 Confocal 100% WARNING! The ImageXpress Confocal HT.ai instrument uses a Class 4 high-power laser light source, which can cause materials to smolder or burn, especially at close range. Dark materials, which absorb heat, and materials such as plastic, paper, and fabric can easily be burned by visible laser beams.
Page 98 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Table 5-7: W# Tab: General Options (continued) Option Description Auto Expose Click to automatically set the exposure time required to obtain the Target Max Intensity. If Auto Expose results in a value of 5 ms or less to achieve this intensity, the software prompts you to activate Bright Sample mode to prevent saturation of the sample.
Page 99 Chapter 5: Configuring Plate Acquisition Options Table 5-8: W# Tab: Autofocus Options (options vary based on Acquisition tab selections) (continued) Option Description Laser and Displays for the first acquisition wavelength (W1) and only if you select Enable Image Laser-Based Focusing and Enable Image-Based Focusing on the Acquisition tab.
Page 100 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Table 5-8: W# Tab: Autofocus Options (options vary based on Acquisition tab selections) (continued) Option Description Image-Based Available for the second or greater acquisition wavelengths and if you select Enable Image-Based Focusing on the Acquisition tab. Specify the Z-Offset from W1 in µm in the Offset field or click Calculate Offset to calculate the offset to use...
Page 101 Chapter 5: Configuring Plate Acquisition Options Table 5-9: W# Tab: Acquisition Options (options vary based on the Series option on the Acquisition tab) (continued) Option Description Acquire Z Single Plane acquires a single plane based on the autofocus options for the Series allows wavelength, as if the Acquire Z Series option were disabled.
Page 102 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Table 5-10: W# Tab: Shading Correction Options Option Description Notes No shading correction or background Not recommended for montages. subtraction is done. Not recommended for assays where intensity of individual objects will be compared across the field of view.
Page 103 Chapter 5: Configuring Plate Acquisition Options Table 5-10: W# Tab: Shading Correction Options (continued) Option Description Notes Background subtraction is performed Not recommended for transmitted Subtraction on the images. No shading correction light images. Only is performed. Background subtraction Not recommended for time lapse removes stray light that is unrelated assays of dim samples.
Page 104 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Table 5-10: W# Tab: Shading Correction Options (continued) Option Description Notes Auto This is a shading algorithm tuned for Not recommended for montages. Correction for transmitted light but can be used for Not recommended for Z-stack fluorescence.
Page 105 Chapter 5: Configuring Plate Acquisition Options Using Legacy Correction Image Shading Note: This section is applicable if you select Legacy Correction as the shading correction method for an acquisition wavelength. See Configuring Acquisition Wavelengths on page Before you do legacy shading correction, acquire the images for each acquisition wavelength and save the images to a stored correction images directory.
Page 106: Configuring Series Acquisition Options
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Configuring Series Acquisition Options The Acquisition tab provides two series acquisition options (you can select one or both): Acquire Time Series acquires images at multiple time points. When you select this option, specify the set of images to acquire at each time point.
Page 107 Chapter 5: Configuring Plate Acquisition Options Configuring Continuous Time Series Acquisitions Use the Timelapse tab to configure time series acquisitions. There are two types of time series acquisitions. This section describes continuous time series acquisition. For discontinuous time series acquisition, see Configuring Discontinuous Time Series Acquisitions on page 109.
Page 108 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 2. Configure the time series acquisition options. Table 5-11: Time Series Acquisition Settings Option Description Number of Specify the total number of time points to acquire. If you change this value, the...
Page 109 Chapter 5: Configuring Plate Acquisition Options 3. In the Interval field, enter the amount of time between the start of an acquisition at one time point and the start of an acquisition at the next time point. Click the corresponding drop-down and select the unit of time measure.
Page 110 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 8. Select the Timelapse tab and do the following to add continuous time points: a. Click the Perform Time Series For drop-down: Select One Well Then The Next for fast kinetic time lapse in a single well (or site).
Page 111 Chapter 5: Configuring Plate Acquisition Options Configuring Z Series Acquisition Options Use the Z Series tab to configure Z series options. To configure Z Series options: 1. Select the Z Series tab. Figure 5-20: Z Series Tab on the Configure Tab Tip: The blue bars represent the planes to acquire for each site imaged.
Page 112 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 2. Configure the Z series acquisition options. Table 5-12: Z Series Acquisition Options Option Description # of Steps The number of planes at a given site or well position for which to take images.
Page 113 Chapter 5: Configuring Plate Acquisition Options 3. To navigate the Z Series for a site or well position and modify the coordinates for the Z motor before you acquire any images, do the following: Figure 5-21: Z Series Diagram Drag the Ruler Zoom slider (below the Z series diagram) or use the mouse scroll wheel to zoom in or zoom out on the diagram.
Page 114 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Drag the Current Position flag on the Z series diagram to update the current position for the Z motor. After you move the flag, you can then click Set T or Set B to set the top and bottom positions.
Page 115: Configuring Journals To Run During Acquisition
Chapter 5: Configuring Plate Acquisition Options Configuring Journals to Run During Acquisition Use the Journals tab to configure journals to run during different stages of the acquisition. You can assign one journal configuration per acquisition step. 1. Select the Acquisition tab. 2.
Page 116 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Table 5-13: Journal Acquisition Steps Option Description Before Each Runs during the acquisition loop, after the illumination is set and focusing is Image complete. After Each Runs during the acquisition loop, after the shutter closes and before images Image save.
Page 117 Chapter 5: Configuring Plate Acquisition Options 5. Click for each acquisition step with a selected check box to display the Select Plate Acquisition Journal dialog. 6. Select a journal file. Figure 5-23: Select Plate Acquisition Journal Dialog 7. Click Open to display the Journal tab. 8.
Page 118: Configuring Post-Acquisition Analysis Options
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Configuring Post-Acquisition Analysis Options Use the Analysis tab to select the analysis to run on a plate after the acquisition completes. If you select this option, the software adds the analysis job to the Auto Run queue in the MDCStore database for analysis either by MetaXpress software that is set to Auto Run mode or by the MetaXpress PowerCore™.
Page 119 Chapter 5: Configuring Plate Acquisition Options Figure 5-25: Analysis Tab (Z Series and 2D Projection Image Only) 4. Click the Analysis drop-down and select the analysis protocol to run post-acquisition. Note: This list includes any application modules, custom modules, or journal assays that you have saved to the MDCStore database.
Page 120 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 6. Select the acquisition time points for which to run the analysis. Table 5-14: Time Point Options - Time Series Acquisition Option Description All Time Points Analyze all acquisition time points. Time Point Range Analyze the acquisition time points that fall within the range.
Page 121: Plate Acquisition Setup Dialog: Configure Tab, Display Tab
Chapter 5: Configuring Plate Acquisition Options Plate Acquisition Setup Dialog: Configure Tab, Display Tab Note: Use the Active Wavelength Preview tool to generate a preview of an acquired image for each wavelength. Use the Display tab to configure the settings for the MetaXpress software desktop appearance such as image size and positions, the Plate Acquisition Setup dialog size and position, and other image properties during acquisition.
Page 122 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Previewing the Acquisition Layout Use the Plate Acquisition Status dialog to preview the display settings. To preview display settings: 1. Select the Display tab. 2. Click Display Acquisition Layout. Individual image widows display where the image will appear for each wavelength and the Plate Acquisition Status dialog displays where any acquisition related dialog will appear during acquisition.
Page 123: Viewing The Summary Panel
Chapter 5: Configuring Plate Acquisition Options Viewing the Summary Panel The Summary panel displays to the right of the Plate Acquisition Setup dialog to list a summary of the current acquisition settings. To view the summary panel: 1. At the bottom of the Plate Acquisition Setup dialog, click Summary >> to display the Summary panel.
Page 124: Saving Plate Acquisition Protocols
Saving Plate Acquisition Protocols You can save a plate acquisition protocol to the database or to a file. If you plan to integrate your ImageXpress Confocal HT.ai system with a robot for automated plate loading, you must save the protocol to a file.
Page 125: Running Plate Acquisition Protocols
Chapter 5: Configuring Plate Acquisition Options Running Plate Acquisition Protocols Use the Run tab in the Plate Acquisition Setup dialog to run the plate acquisition protocol. After you load a protocol, configure the wells and sites for which to acquire a data, and verify that the current acquisition settings are appropriate, you can run the protocol and acquire the plate data.
Page 126 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 6. Do the following to make last minute tweaks as needed: Click the blue wavelength name to display the corresponding W# Wavelength tab where you adjust wavelength settings. See Configuring Acquisition Wavelengths on...
Page 127: Chapter 6: Customizing The Metaxpress Software
Chapter 6: Customizing the MetaXpress Software A powerful feature of the MetaXpress software is the ability to customize the operation of the software for individual users or groups of users. Various objectives and workflows call for customized settings that can be switched as needed. Programs within the MetaXpress Software Suite, such as the Meta Imaging Series Administrator Software and the Create Taskbar command, let you create settings that match the needs of your users.
Page 128: Users And Groups In The Meta Imaging Series Administrator Software
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Users and Groups in the Meta Imaging Series Administrator Software Use the Meta Imaging Series Administrator Software commands to define and configure settings for individual users and groups in the MetaXpress software. There are two modes for...
Page 129 Chapter 6: Customizing the MetaXpress Software Creating an Offline Version of the MetaXpress Software We recommend creating an offline group for MetaXpress software users. This offline group does not include hardware settings and is useful for analysis of acquired images. Since the offline group does not include hardware settings, it does not attempt to establish communication with the other MetaXpress software components.
Page 130 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Creating MetaXpress Software Group Icons and Adding Them to the Windows Desktop After you create the MetaXpress software offline group and add users with the previous procedure, use the Create Icons command to create icons for the new group. This command installs shortcuts for new groups to the MetaXpress folder on the Windows desktop.
Page 131: Custom Toolbars And Taskbars
Chapter 6: Customizing the MetaXpress Software Custom Toolbars and Taskbars After you configure groups, you can create or modify custom toolbars and taskbars to include specific combinations of tools and commands. Customizing Toolbars Use the Configure Drop-ins/Toolbars command to add menu commands to toolbars, move commands from one toolbar to another, and add journals to or remove journals from toolbars.
Page 132 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Creating Taskbars Use Taskbars to access frequently used commands and journals. Each taskbar can have up to 48 buttons in a configuration of rows and columns. You can mix and match journals, commands, or other taskbars within the same taskbar.
Page 133 Chapter 6: Customizing the MetaXpress Software 8. If needed, click Undo to undo the last action or click Clear Button to clear an item from the active button. 9. Click Rename Taskbar to display the Rename Taskbar dialog where you change the name of the taskbar.
Page 134: Default Paths For Data
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Default Paths for Data Use the Configure Default Paths dialog to change the default file paths for each group or user. You can modify these paths so that the users on the system have their own dedicated data folders.
Page 135: Chapter 7: Expansion Solution Options
Chapter 7: Expansion Solution Options You can upgrade the ImageXpress Confocal HT.ai system to include the following expansion solution options: Environmental Control Option, see below Transmitted Light Option, see page 162 Water Immersion Option, see page 179 Contact your Molecular Devices representative to discuss adding expansion solution options to your system.
Page 136 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Items Included in the Installation The following hardware components are included in an ImageXpress Environmental Control option installation: ImageXpress Systems Power and Options Controller. See Figure 7-1. Warm air hose and CO tubing. See...
Page 137 Chapter 7: Expansion Solution Options Figure 7-3: Warm Air Hose and CO Tube on the Back of the ImageXpress Confocal HT.ai System Figure 7-4: Water Reservoir Figure 7-5: Live-Cell Sealing Ring 5084773 A...
Page 138 After you release «, the current temperature in the chamber is displayed. Note: For the ImageXpress Confocal HT.ai 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 139 Chapter 7: Expansion Solution Options Setting Up the Water Reservoir The water reservoir adds humidity to the air flow supplied to the environmental enclosure. Do one of the following: Filling the Water Reservoir Outside the Instrument, see below Filling the Water Reservoir Inside the Instrument, see page 143 Filling the Water Reservoir Outside the Instrument You will need the following: Syringe...
Page 140 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 4. Remove the lock screw from the top access panel. Figure 7-8: Top Access Panel Lock Screw 5. Remove the top access panel surrounding the top door and open the upper hinged side door for more access.
Page 141 Chapter 7: Expansion Solution Options 6. Locate the water reservoir through the top opening. Figure 7-10: Environment Control Water Reservoir Table 7-2: Environment Control Water Reservoir Description Item Description Luer Locks Water Sensors Wires Water Sensor 7. Disconnect the CO /air tubing from the luer locks on both side of the water reservoir.
Page 142 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 13. Reconnect the wires to the sensors on the green luer lock side. The color of the sensor wire is irrelevant when connecting to the sensor socket. Either color of sensor wire can connect to either sensor socket.
Page 143 Chapter 7: Expansion Solution Options Filling the Water Reservoir Inside the Instrument You will need the following: Syringe 250 mL (8.5 oz) deionized water, preferably sterilized To fill the water reservoir inside the instrument: 1. In the MetaXpress software, do one of the following to open the instrument top door: From the simplified menu, click Screening >...
Page 144 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 4. Remove the lock screw from the top access panel. Figure 7-14: Top Access Panel Lock Screw 5. Remove the top access panel surrounding the top door and open the upper hinged side door for more access.
Page 145 Chapter 7: Expansion Solution Options 6. Locate the water reservoir through the top opening. Figure 7-16: Environment Control Water Reservoir Table 7-4: Environment Control Water Reservoir Description Item Description Luer Locks Water Sensors Wires Water Sensor 7. Open the upper hinged side door for more access. 8.
Page 146 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Setting Up the CO Tank A pre-mixed CO tank with a regulator must be set up and connected to the CO inlet on the ImageXpress Systems Power and Options Controller, which controls the flow rate of CO delivered to the water reservoir within the environmental enclosure.
Page 147 Chapter 7: Expansion Solution Options 5. Remove the lock screw from the top access panel. Figure 7-18: Top Access Panel Lock Screw 6. From the top of the instrument, lift the top access panel surrounding the top door and remove it. Keep the instrument top door closed. Figure 7-19: Top Access Panel Around the Top Door On and Off 7.
Page 148 ImageXpress Confocal HT.ai High-Content Imaging System 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 and temperature levels.
Page 149 Chapter 7: Expansion Solution Options Before You Run an Experiment Check the water reservoir level. Refill as needed. Check the CO supply. Refill as needed. Allow the instrument to warm up for at least two hours. The current temperature in the chamber is displayed on the front of the controller.
Page 150 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Environmental Control Software The MetaXpress software controls the ImageXpress Confocal HT.ai instrument screening and monitors the system’s environmental parameters. You can check Environment Control settings and connections in the software. To check the Environment Control settings: 1.
Page 151 Chapter 7: Expansion Solution Options Performing Time Lapse Experiments The ImageXpress Environmental 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 time lapse acquisitions, the time starts when the first well is acquired.
Page 152 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 4. Select the Timelapse tab. Figure 7-23: Plate Acquisition Setup Dialog, Timelapse Tab 5. In the Number of time points field, enter the number of time points. 6. Click the Perform Time Series For drop-down: Select All Selected Wells for long time lapse experiments.
Page 153 Chapter 7: Expansion Solution Options 9. If applicable, select each W# Wavelength tabs. Figure 7-25: Plate Acquisition Setup Dialog, Wavelength 1 Tab 10. Click the Timelapse drop-down: Select At All Time Points to acquire all time points for the wavelength. Select At Start of Experiment to acquire only the first time point for the wavelength.
Page 154 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Reviewing Time Lapse Data As with other ImageXpress System data, time lapse data displays in the Review Plate Data dialog. To review the time lapse data: 1. In the MetaXpress software, do one of the following to display the Review Plate Data dialog: From the simplified menu, click Screening >...
Page 155 Chapter 7: Expansion Solution Options 8. In the image window, use the navigation buttons above the image pane to view each time point as a plane in the stack. Figure 7-27: Stack of Images with Navigation Buttons Above the Image Pane Note: In a time lapse experiment, the time starts when the first well is acquired.
Page 156 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Making Time Lapse Movies To make a movie: 1. Before you begin, load time lapse images and create a stack. 2. Select the image window of the stack on the MetaXpress software desktop.
Page 157 Chapter 7: Expansion Solution Options Figure 7-29: CO Tubing Connections on the Water Reservoir Table 7-5: CO Tubing Connections on the Water Reservoir Descriptions Item Description Input tube connection Output tube connection CAUTION! Do not let the second stretch of tubing that leaves the water reservoir to the stage area come into contact with the water in the reservoir.
Page 158 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Cleaning the Water Reservoir Clean the water reservoir as needed when you see dirt or mold in the reservoir. Requirements: Phillips screwdriver 70% ethanol 250 mL (8.4 oz) deionized water, preferably sterilized To clean the water reservoir: 1.
Page 159 Chapter 7: Expansion Solution Options 4. Remove the lock screw from the top access panel. Figure 7-31: Top Access Panel Lock Screw 5. Remove the top access panel surrounding the top door and open the upper hinged side door for more access. Figure 7-32: Top Access Panel Removed 5084773 A...
Page 160 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 6. Locate the water reservoir through the top opening. Figure 7-33: Environment Control Water Reservoir Table 7-7: Environment Control Water Reservoir Description Item Description Luer Locks Water Sensors Wires Water Sensor 7. Disconnect the CO /air tubing from the luer locks on both sides of the water reservoir.
Page 161 Chapter 7: Expansion Solution Options 15. Reconnect the wires to the sensors on the green luer lock side. Note: The color of the sensor wire is irrelevant when connecting to the sensor socket. Either color of sensor wire can connect to either sensor socket. Figure 7-34: Water Level Sensor Wire Sockets Figure 7-35: Water Level Sensor Wires Inside the Instrument 16.
Page 162: Transmitted Light Option
See Figure 7-37. Phase alignment lens. Located in the emission filter wheel and is placed in position using the MetaXpress software for the ImageXpress Confocal HT.ai system. Hinge. Allows robotic access to the plate if necessary. See Figure 7-38.
Page 163 Chapter 7: Expansion Solution Options Figure 7-36: Transmitted Light Option (Detailed View) Figure 7-37: Transmitted Light Shutter 5084773 A...
Page 164 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Figure 7-38: Hinge Release Knob and Hinge Table 7-8: Hinge Release Knob and Hinge Description Item Description Hinge Release Knob (in place) Hinge Hinge Release Knob (removed) 5084773 A...
Page 165 Chapter 7: Expansion Solution Options 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 166 ImageXpress Confocal HT.ai High-Content Imaging System User Guide The following example sets the lamp power to 20% of maximum. Figure 7-41: Configure Illumination Dialog: Transmitted Light TL now displays as an option in the Plate Acquisition Setup dialog from the Active Wavelength drop-down and on each W# Wavelength tab from the Illumination drop-down.
Page 167 Condenser Focus Control Condenser Adjustment Screws (Aperture Lateral Adjustment Control) Condenser Turret Field Stop Slider (Condenser Aperture Diaphragm Control) Note: For the ImageXpress Confocal HT.ai system, make sure that the instrument is in widefield mode before setting up Koehler illumination. 5084773 A...
Page 168 ImageXpress Confocal HT.ai High-Content Imaging System User Guide To set up the ImageXpress Transmitted Light option for Koehler Illumination: 1. Load the sample (multi-well plate or slide). 2. Select a low magnification objective such as the 10x Plan Apo or a 4x objective.
Page 169 Chapter 7: Expansion Solution Options 16. Click Focus to confirm proper illumination. If you see dark edges in your field of view, repeat step 13. Figure 7-45: Adjustment of the Field Aperture Diaphragm Comparison 5084773 A...
Page 170 The phase ring alignment procedure requires a special lens to be placed in position before starting the phase ring adjustment process. On the ImageXpress Confocal HT.ai system, you must install the phase ring adjustment lens in Position 8 of the emission wheel to perform the phase ring adjustment process. After the adjustment process, you must remove the phase ring adjustment lens and replace it with the original filter.
Page 171 Chapter 7: Expansion Solution Options 2. Rotate the condenser turret so that the A position is selected (at the front). Figure 7-46: Condenser Turret with Position A at the Front 3. If Koehler illumination has not been set up, turn the adjustment ring, which is located below the arm of the Transmitted Light tower, to the middle of the aperture range.
Page 172 The image of the phase ring might be off-center. This is acceptable if most of the image is visible. Tip: If the phase ring is not visible at all, contact Molecular Devices Technical Support. See Obtaining Support on page 237.
Page 173 Chapter 7: Expansion Solution Options 12. Adjust the position of the Z-stage to obtain a reasonably sharp image of the objective phase ring. Figure 7-50: In Focus Image of Phase Contrast Ring 13. Use the region tools in the software to mark the position of the phase ring. Use the rectangular region tool to make sure that you capture both horizontal and vertical axes.
Page 174 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 16. Snap an image to show the condenser annulus ring. You might need to adjust the exposure time. Figure 7-52: Image of Condenser Ring with the Loaded Region 17. In either the Acquire dialog or the Plate Acquisition and Control dialog, click Live Mode.
Page 175 Chapter 7: Expansion Solution Options 22. In the MetaXpress software, click Stop Live. The lens you use for phase ring adjustment automatically moves out of the optical path when you select an emission wavelength for the acquisition. 23. In the Configure Illumination dialog, verify that the IXConfocal Module Emission Filter setting is restored to its normal setting, such as Empty.
Page 176 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Performing Transmitted Light Experiments Do the following procedure if you are running an experiment with Transmitted Light as one of the selected wavelengths. To run a Transmitted Light experiment: 1. If you run a phase-contrast experiment, make sure to select a phase-contrast objective.
Page 177 Chapter 7: Expansion Solution Options Replacing the Transmitted Light Bulb The ImageXpress Transmitted Light option uses a replaceable 12-volt halogen light bulb. See Optional Parts and Accessories on page 249. WARNING! The surface of the halogen lamp gets very hot during operation and remains hot for up to 20 minutes after operation.
Page 178 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 4. Face the light source from the rear and identify the bulb and the lever. Figure 7-56: Bulb and Lever 5. Put on cotton gloves to handle the bulb. CAUTION! Never touch the bulb with bare hands.
Page 179: Water Immersion Option
CAUTION! Do not remove or change the water immersion objectives that are installed within the instrument. A Molecular Devices Field Service Engineer must install and remove water immersion objectives. Do not touch the leak detection sensor, which is the brass color ring around the top of the objective below the cap.
Page 180 Water Immersion Objectives CAUTION! Do not remove or change the water immersion objectives that are installed within the instrument. A Molecular Devices Field Service Engineer must install and remove water immersion objectives. The Water Immersion option can include up to two of the water immersion objectives listed in the following table.
Page 181 Relief valve to maintain constant pressure CAUTION! Do not move the water immersion controller to a location that is above or below where it is installed by the Molecular Devices Field Service Engineer. To ensure high-performance acquisition with water immersion, the Field Service Engineer calibrates the water dispense rate.
Page 182 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Performing Experiments with the Water Immersion Option The process for acquiring images with water immersion objectives is essentially the same as the process for acquiring images with air objectives. This section describes the additional...
Page 183 Chapter 7: Expansion Solution Options Setting Up Plate Acquisition to Use a Water Immersion Objective In the MetaXpress software, you configure an acquisition using a water immersion objective the same way that you would configure an acquisition using an air objective. When using a water immersion objective, the Plate Acquisition Setup dialog provides: A water immersion status icon beneath the plate map.
Page 184 ImageXpress Confocal HT.ai High-Content Imaging System User Guide The following table describes how the WI Status icon indicates the status of the water immersion system. Table 7-13: WI Status descriptions Icon System Status The water immersion system and settings are appropriately configured.
Page 185 Chapter 7: Expansion Solution Options Confirming Water Immersion Status The software automates most water immersion activity. The Water Immersion Control dialog is provided primarily for informational purposes. Figure 7-60: Water Immersion Control Dialog To open the Water Immersion Control dialog, do one of the following: In the Plate Acquisition Setup dialog, click WI Status.
Page 186 If you plan to store the instrument, you plan to move the instrument, or you do not intend to use the water immersion system for a long period of time, contact Molecular Devices for instructions.
Page 187 To test that the vacuum line is not clogged, click Test Vacuum Line. c. To test that water is dispensed correctly, click Test Dispense. If a message continues to indicate an error or issues persist, contact Molecular Devices. 5084773 A...
Page 188 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 5084773 A...
Page 189: Chapter 8: Maintenance
To avoid personal injury or damage to the instrument during maintenance procedures, observe the following precautions: Some procedures require you to fully shut down the ImageXpress Confocal HT.ai system and unplug the power cable. See Shutting Down the System on page If the MetaXpress software is running, exit the software before you turn off the instrument.
Page 190: Routine Maintenance
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Routine Maintenance To ensure optimal operation of the instrument, perform the following routine maintenance procedures as needed: Wipe off visible dust from exterior surfaces with a lint-free cloth to avoid dust build-up on the instrument.
Page 191 Chapter 8: Maintenance Cleaning the ImageXpress Confocal HT.ai System The following procedure is designed to clean the plate-loading region of the instrument without damaging the internal components of the imaging system. Before you clean the instrument, read and follow the Maintenance Precautions on page 189.
Page 192: Light Source Maintenance
Replacing the Confocal Disk Box In the ImageXpress Confocal HT.ai system, the Confocal disk box is installed in the Confocal module housing within the lower enclosure of the instrument. There are three Confocal disk box configurations. The standard unit contains a single 60 µm pinhole Confocal disk for basic Confocal protocols.
Page 193 Chapter 8: Maintenance The following sections describe how to replace the Confocal disk box: Removing the Disk Box, see below Installing the Disk Box, see page 194 Testing the Disk Box, see page 195 Removing the Disk Box Use this procedure to change from one type of Confocal disk box to another type of Confocal disk box, or to replace a Confocal disk box.
Page 194 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Installing the Disk Box Note: Before you begin, follow the steps in the previous sections to remove the disk box. To install a disk box: 1. Use bulb duster to blow visible particulates off the disk box lens and window.
Page 195 Chapter 8: Maintenance Testing the Disk Box Use this procedure to verify confocal disk box is installed and functioning correctly. To test a disk box: 1. In the MetaXpress software, do one of the following to open the instrument top door: From the simplified menu, click Screening >...
Page 196 Click Show Live for a live view. WARNING! The external light source for the ImageXpress Confocal HT.ai system is a Class 4 high-power laser product. Prolonged exposure can cause plate materials to smolder or burn. If live-mode exposure is longer than 30 seconds, the software automatically closes the laser shutter.
Page 197 Chapter 8: Maintenance 13. Go back to the Component Control dialog. Figure 8-8: Component Control Dialog (Running) 14. Click the Position drop-down and select any Running position. Note: Running positions are odd numbers. 15. Wait about 2 minutes for the disk to start spinning. 16.
Page 198: Creating Hardware Profiles
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Creating Hardware Profiles A hardware profile is a system settings file to which the software refers to identify which hardware and configurations are available for use. To create a new hardware profile: 1.
Page 199 Chapter 8: Maintenance 3. Click Configure Hardware to display the Configure Hardware dialog. Figure 8-12: Configure Hardware Dialog 4. Click Create New Setting to display the Create New Setting dialog. Figure 8-13: Create New Setting dialog 5. In the Setting Name field, enter the setting name. 6.
Page 200: Filter Maintenance
Mode to prevent errors or missing settings. Filter Maintenance Whenever you replace or add to the optical components in the ImageXpress Confocal HT.ai system, you must also enter the filter information into the MetaXpress software to reflect the new hardware configuration.
Page 201 Chapter 8: Maintenance Replacing Emission Filters The emission filters are installed in a wheel within the confocal module housing. The accessory kit includes a specialized maintenance tool that you can use to replace the emission filter. You may also need the following basic tools required to replace a filters: Powder-free gloves Flashlight Bulb duster...
Page 202 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 5. In the Claimed Devices list, under ImageXpress Confocal Module, select IXConfocal Module Emission Wheel and click Settings to display the IXConfocal Module Emission Wheel Settings dialog. Figure 8-17: IXConfocal Module Emission Wheel Settings Dialog 6.
Page 203 Chapter 8: Maintenance 11. Open the filter access door on the left of the confocal module housing. The slot for the emission filter is on the left. Figure 8-19: Confocal Module Housing with Filters Access Door Open 12. Use a flashlight to look into the emission access slot to get familiar with the tool alignment groove and verify that the you can see the rim of the filter.
Page 204 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Installing Emission Filters Note: Before you begin, follow the steps in the previous section to remove the emission filter. To install an emission filter: CAUTION! To protect the emission filter, load the emission tool with the emission filter over a table or other work surface.
Page 205 Chapter 8: Maintenance 9. If the filter is a different kind than the one you removed, in the IXConfocal Module Emission Wheel Settings dialog, in the corresponding Position Labels field, enter the new filter name. Figure 8-22: IXConfocal Module Emission Wheel Settings Dialog 10.
Page 206 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Editing Filter Settings The ImageXpress Confocal HT.ai system uses emission filters and dichroic mirror filters. If needed, you can edit the settings for the emission filters in the Meta Imaging Series Administrator Software.
Page 207 Chapter 8: Maintenance Editing Illumination Settings To edit illumination settings: 1. 1. In the MetaXpress software, do one of the following to display the Configure Illumination dialog: From the simplified menu, click Control > Devices > Configure Illumination. From the standard menu, click Devices > Configure Illumination. Figure 8-24: Configure Illumination Dialog 2.
Page 208 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 4. Set up other illumination settings if needed. The value in the Wavelength field should match the center wavelength for the emission filter. CAUTION! Never set more than one 89 North shutter to Active.
Page 209: Objective Maintenance
HT.ai system, you must physically change or add the component and then enter the changes in the software to reflect the new hardware configuration. CAUTION! A Molecular Devices Field Service Engineer must install and remove water immersion objectives. The Water Immersion option supports up to two water immersion objectives and they must be installed in the first and/or fourth objective slots.
Page 210 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Correct Objective Placement Whenever possible, Molecular Devices installs objectives with correction collars in the outer positions 1 and 4. The outer positions provide easier access through the side panels to adjust the correction collar.
Page 211 Cleaning Objectives Clean the objective when debris or contaminants collect on an objective. CAUTION! You must contact Molecular Devices Technical Support to schedule a Field Service Engineer to install or remove a water immersion objective. Before you clean an objective, read and follow the Maintenance Precautions on page 189.
Page 212 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Using Oil-Immersion Objectives You can use oil-immersion objectives with the ImageXpress Confocal HT.ai system for research-mode imaging. For ordering information, contact your sales representative. Note: We do not recommend using an oil-immersion objective to scan an entire plate.
Page 213 5. Locate the correction collar on the objective. Loosen or remove an air objective to locate the correction collar, if needed. CAUTION! You must contact a Molecular Devices Field Service Engineer to remove a water immersion objective. 6. Rotate the correction collar to its new setting. If needed, replace or tighten the air objective.
Page 214: Installing Objectives
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Installing Objectives The objectives in the ImageXpress Confocal HT.ai system are mounted in a four-position linear selector. Objectives are very delicate components. Use special care when you handle objectives. Also see Compatible Objectives on page 243.
Page 215 Chapter 8: Maintenance 18. Do one of the following to display the Configure Magnification dialog: From the simplified menu, click Control > Devices > Configure Magnification. From the standard menu, click Devices > Configure Magnification. 19. Enter the new objective information. See Updating Magnification Settings on page 230.
Page 216 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Editing Objective Settings in the Meta Imaging Series Administrator Software To edit objective settings: 1. Click Start > MetaXpress 6 > Meta Imaging Series Administrator to display the Meta Imaging Series Administrator dialog.
Page 217 Chapter 8: Maintenance 9. In the Installed Devices field, select ImageXpress Micro and then click Settings to display the ImageXpress Micro Settings dialog. 10. Select the Parameter Group #1 tab. 11. For a Plate Bottom Reference and XY reference, select the Maintenance Mode check box. 12.
Page 218: Configuring Parfocality After Changing Objectives
MetaXpress Objective dialog in the Meta Imaging Series Administrator Software. You can configure parfocality with or without the taskbar. CAUTION! You must contact Molecular Devices Technical Support to schedule a Field Service Engineer to install or remove a water immersion objective.
Page 219 11. Click Show Live to display a Live image window. WARNING! The external light source for the ImageXpress Confocal HT.ai system is a Class 4 high-power laser product. Prolonged exposure can cause plate materials to smolder or burn. If live-mode exposure is longer than 30 seconds, the software automatically closes the laser shutter.
Page 220 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 12. Do one of the following to display the Move Stage to Absolute Position dialog: From the simplified menu, click Control > Devices > Move Stage to Absolute Position. From the standard menu, click Devices > Move Stage to Absolute Position.
Page 221 Chapter 8: Maintenance 15. In the Configure Magnification dialog, click the Setting drop-down and select the objective with the next highest numerical aperture (NA) that is in position #2. Repeat the following steps for each objective and write down the focus values for each. You will use these values to determine the parfocality for each objective.
Page 222: Determining The Plate Bottom Reference Point After Changing The Reference Objective
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Determining the Plate Bottom Reference Point After Changing the Reference Objective Note: Do this procedure only if you replace the reference objective. If you did not replace the reference objective, see Entering the Focus Objective Values in the Meta Imaging Series Administrator Software on page 228.
Page 223 Chapter 8: Maintenance Finding the Plate Bottom Reference Objective and Reference Point Before you start this procedure, make sure that the MetaXpress software is in maintenance mode. See Editing Objective Settings in the Meta Imaging Series Administrator Software on page 216.
Page 224 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 13. In the Current Position: X field, enter 64900. 14. In the Current Position: Y field, enter 42200, and then press Enter to move to the approximate center of the plate. 15. Do one of the following to display the Focus dialog: From the simplified menu, click Control >...
Page 225 Chapter 8: Maintenance 23. In the Focus dialog, on the Auto Focus tab, click Preview Pass to display a Preview Pass image window that displays a graph of focus intensities compared to Z-position. Figure 8-33: Preview Pass Image Window The top of the red peak represents the brightest pixel of the preview pass. The top of the green peak represents the highest focus score.
Page 226 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 25. Write down the Z-position for the center focus position in the following table. Table 8-4: Plate Bottom Reference Points Position X Value Y Value Z Value Center 64900 42200 Upper-Left 8900...
Page 227 Chapter 8: Maintenance Entering the Plate Bottom Reference Objective Value in the MetaXpress.ref Configuration File Note: Only do this procedure if you need to complete Finding the Plate Bottom Reference Objective and Reference Point on page 223. To enter the value for the position of the plate bottom reference objective in the MetaXpress.ref configuration file: 1.
Page 228: Entering The Focus Objective Values In The Meta Imaging Series Administrator Software
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Entering the Focus Objective Values in the Meta Imaging Series Administrator Software Turn off maintenance mode before you do the steps in this section. See Entering the Focus Objective Values in the Meta Imaging Series Administrator Software on page 228.
Page 229 Chapter 8: Maintenance 16. If you did the steps to Finding the Plate Bottom Reference Objective and Reference Point on page 223: a. In the Installed Devices field, select ImageXpress Micro Z. b. Click Settings to display the ImageXpress Micro Z Settings dialog. c.
Page 230: Updating Magnification Settings
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Updating Magnification Settings Before you begin this procedure, make sure that the MetaXpress software is NOT in maintenance mode. See Entering the Focus Objective Values in the Meta Imaging Series Administrator Software on page 228.
Page 231 Chapter 8: Maintenance Calibrating Distances Turn off maintenance mode before you do the steps in this section. See Entering the Focus Objective Values in the Meta Imaging Series Administrator Software on page 228. To calibrate distances: 1. Do one of the following to display the Calibrate Distances dialog: From the simplified menu, click Measure >...
Page 232 4. In the X and Y fields, enter the calibration values for the new objective. X and Y values must match. You can use the following estimated values for ImageXpress Confocal HT.ai system calibration settings: Note: To measure pixel sizes more accurately, in the IX Taskbar, click System Maintenance and then click Measure Pixel Sizes.
Page 233: Updating Legacy Shading Correction Settings
Chapter 8: Maintenance Updating Legacy Shading Correction Settings Note: This procedure is required if you select the Legacy Correction check box for one or more acquisition wavelengths. All legacy correction images should be acquired using the widefield acquisition mode. If you select Legacy Correction as the Shading Correction method for an acquisition wavelength, you need the shading correction image files for each magnification and illumination setting.
Page 234 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Importing the Main Taskbar Journal Suite The IX Taskbar is a collection of tools that enhance and streamline common imaging workflows. Do the following procedure only if the Main Taskbar is not installed on the computer. If the...
Page 235 Chapter 8: Maintenance 15. Navigate to C:\MX6\Taskbars or the location in your custom installation. 16. Select the Main Taskbar.jtb file. 17. Click Open to display the Main Taskbar dialog. Figure 8-40: Main Taskbar Dialog 18. Click Run IX Taskbar Installer to display the IX Taskbar Installer dialog. Figure 8-41: IX Taskbar Installer 19.
Page 236: Turning Off The Status Indicator Lights
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Running the Shading Correction Journal from the Main Taskbar To run the shading correction journal from the Main taskbar: 1. From the Main taskbar, select System Maintenance and then click Set up Shading Correction.
Page 237: Obtaining Support
Our Support website—www.moleculardevices.com/service-support—describes the support options offered by Molecular Devices, including service plans and professional services. It also has a link to the Molecular Devices Knowledge Base, which contains documentation, technical notes, software upgrades, safety data sheets, and other resources.
Page 238 Gathering Support Information Before you contact Molecular Devices for support, you need the following information available to help the Technical Support personnel troubleshoot the problem: The steps that led up to the occurrence of the problem The settings of any dialogs used when the problem occurred...
Page 239 5. Click File > Exit to close the AxoTrace window. 6. Start the MetaXpress software. 7. Turn off the ImageXpress Confocal HT.ai system and then turn it back on. The light source can remain on during this step. 8. Reproduce the error.
Page 240 6. In the Name field enter the file name. 7. Navigate to the location to save the file. 8. Click Save to display the report in .pdf format. 9. Send the report file to the Molecular Devices support personnel as an email attachment. 5084773 A...
Page 241: Appendix A: Instrument Specifications
For additional specifications, refer to the ImageXpress Confocal HT.ai Pre-Installation Guide . WARNING! If the instrument is used in a manner not specified by Molecular Devices, the protection provided by the equipment might be impaired. The ImageXpress Confocal HT.ai system is an Equipment Class 1 product that relies on protective earth grounding for safe operation.
Page 242 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Instrument Dimensions Figure A-1: Instrument Height, Width, and Length Figure A-2: Distance Between the Outside Edges of the Instrument Feet 5084773 A...
Page 243: Appendix B: Compatible Objectives
Appendix B: Compatible Objectives The following table details the Nikon objectives that are compatible with the ImageXpress Confocal HT.ai system. It also provides plate compatibility information and theoretical calculation of confocal performance for the objectives you use in the system.
Page 244 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Table B-1: Nikon Objectives Compatible with the ImageXpress Confocal HT.ai System and Settings (continued) Objective Magnification and Type Phase Numerical Working (Molecular Devices Part Number) Contrast Aperture (NA) Distance Plate Compatibility 20x Plan Apo Lambda 0.75...
Page 245 Appendix B: Compatible Objectives Table B-1: Nikon Objectives Compatible with the ImageXpress Confocal HT.ai System and Settings (continued) Objective Magnification and Type Phase Numerical Working (Molecular Devices Part Number) Contrast Aperture (NA) Distance Plate Compatibility 60x Plan Apo Lambda Oil 1.40...
Page 246 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Table B-2: Theoretical Calculation of Confocal Performance with Compatible Objectives Optical Optical 60 µm Pinhole 42 µm Pinhole 50 µm Thickness Thickness Diameter in Diameter in Slit Width in using 60 µm using 42 µm Objective Airy Units Airy Units...
Page 247: Appendix C: Filter Specifications And Calibration Compatibility
Appendix C: Filter Specifications and Calibration Compatibility Shading Correction Calibration Filter Wavelengths Plate Slide DAPI Excitation: 405/20 nm Pink Green (GP-8) Emission: 452/45 nm Dichroic: FF409-Di03 Excitation: 445/20 nm Pink Yellow (GP-11) Emission: 483/32 nm Dichroic: ZT 445/520 FITC Excitation: 467.5/21 nm Pink Yellow (GP-11) Emission: 520/28 nm...
Page 248 ImageXpress Confocal HT.ai High-Content Imaging System User Guide 5084773 A...
Page 249: Appendix D: Optional Parts And Accessories
Appendix D: Optional Parts and Accessories This section describes the items available from Molecular Devices for the ImageXpress Confocal HT.ai system. For a list of compatible Nikon objectives, see Compatible Objectives on page 243. Part Number Description 5039730 Confocal Single Disk Unit, including: 60 µm pinhole disk for basic confocal imaging...
Page 250 ImageXpress Confocal HT.ai High-Content Imaging System User Guide Part Number Description E1005 Breathe Seal for Cell & Tissue Culture, sterile, packed in tamperproof bags of 25; pack of 50 5004555 Immersion Oil, 8 cc, MXA20233 1-2600-0865 Replacement Live Cell Plate Sealing Ring...
Page 251: Appendix E: File Privileges For The Metaxpress Software
Appendix E: File Privileges for the MetaXpress Software This appendix describes which directories must be accessible to MetaXpress software administrators and users. The following information assumes that the MetaXpress software is installed to the default path of C:\MX6. Note: There are no restrictions as to where the software is installed. You can install the software to C:\Program Files\MX6.
Page 252 .zip file in the C:\Backup\ folder. If the software configuration or instrument settings get corrupted, you can restore the settings from the backup archives. Contact Molecular Devices Technical Support for assistance. C:\Shading Images...
Page 253: Appendix F: Robotic Plate Handling
Appendix F: Robotic Plate Handling If your ImageXpress Confocal HT.ai system has a robotic plate handler integrated with the base imaging system, there are additional hardware and user procedures. The robotic plate handler is integrated with the ImageXpress Confocal HT.ai system so that plates can be scanned for barcodes, loaded onto the imaging system where images are acquired, and then returned to a home location.
Page 254: Verifying External Control Settings
ImageXpress Confocal HT.ai High-Content Imaging System User Guide Verifying External Control Settings If you have a robot attached to the ImageXpress Confocal HT.ai system, you need to confirm that the External Control settings in the Meta Imaging Series Administrator Software are enabled and that the correct COM port is selected.
Page 255: Appendix G: Electromagnetic Compatibility
Appendix G: Electromagnetic Compatibility Regulatory for Canada (ICES/NMB-001:2020) This ISM device complies with Canadian ICES-001. Cet appareil ISM est confomre à la norme NMB-001 du Canada. ISM Equipment Classification (Group 1, Class A) This equipment is designated as scientific equipment for laboratory use that intentionally generate and/or use conductively coupled radio-frequency energy for internal functioning, and are suitable for use in all establishments, other than domestic and those directly connected to a low voltage power supply network which supply buildings used for domestic...
Page 256 The trademarks used herein are the property of Molecular Devices, LLC or their respective owners. ©2021 Molecular Devices, LLC. Specifications subject to change without notice. Patents: www.moleculardevices.com/patents All rights reserved. FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES...

Molecular Devices ImageXpress Confocal HT.ai User Manual

Chapter 1: Imagexpress Confocal Ht.ai System

Chapter 2: Using the Imagexpress Confocal Ht.ai System

Chapter 3: System Installation and Testing

Chapter 4: Preparing for Acquisition

Chapter 5: Configuring Plate Acquisition Options

Chapter 6: Customizing the Metaxpress Software

Chapter 7: Expansion Solution Options

Chapter 8: Maintenance

Appendix A: Instrument Specifications

Appendix B: Compatible Objectives

Appendix C: Filter Specifications and Calibration Compatibility

Appendix D: Optional Parts and Accessories

Appendix E: File Privileges for the Metaxpress Software

Appendix F: Robotic Plate Handling

Appendix G: Electromagnetic Compatibility

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