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Takara Bio ICELL8 User Manual

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Takara Bio USA, Inc.
ICELL8® Imaging
System User Manual
Cat. No. 640000
(071919)
Takara Bio USA, Inc.
1290 Terra Bella Avenue, Mountain View, CA 94043, USA
U.S. Technical Support:
technical_support@takarabio.com
United States/Canada
Asia Pacific
800.662.2566
+1.650.919.7300
Europe
Japan
+33.(0)1.3904.6880
+81.(0)77.565.6999
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  • Page 1 Takara Bio USA, Inc. ICELL8® Imaging System User Manual Cat. No. 640000 (071919) Takara Bio USA, Inc. 1290 Terra Bella Avenue, Mountain View, CA 94043, USA U.S. Technical Support: technical_support@takarabio.com United States/Canada Asia Pacific Europe Japan Page 1 of 18 800.662.2566...

  • Page 2: Table Of Contents

    Figure 1. Software interface for cell imaging......................... 4 Figure 2. Workflow diagram depicting single-cell isolation and imaging steps..............5 Figure 3. ICELL8 Imaging System Components........................6 Figure 4. Starting the Mercury Burner............................ 7 Figure 5. Starting the Camera..............................7 Figure 6.

  • Page 3: Preface

    About this Manual This manual provides instructions for safe and successful operation of the imaging system included with the ICELL8 Single-Cell System (Cat. No. 640000). Please be sure to review the information in this manual thoroughly before using the equipment.

  • Page 4: Introduction

    ICELL8 chip. The Micro-Manager software takes 288 images through DAPI and Texas Red filters from 144 fields of view (6 x 6 wells in each field of view) in approximately 7 minutes. The cell specimens can be quickly returned to cold storage while the images are analyzed.

  • Page 5: Figure 2. Workflow Diagram Depicting Single-Cell Isolation And Imaging Steps

    ICELL8® Imaging System User Manual Workflow Diagram The imaging system described in this user manual is provided as a component of the ICELL8 Single-Cell System. For detailed information about other ICELL8 components, as well as compatible kits and reagents, please refer to corresponding documents.

  • Page 6: Figure 3. Icell8 Imaging System Components

    Micro-Manager software: Micro-Manager is open source software, created by the Vale Laboratory at UCSF and adapted for use with the ICELL8 Imaging System. Micro-Manager software controls the motorized stage, optical filters, and camera. It also acquires and manages microscope images.

  • Page 7: Startup

    ICELL8® Imaging System User Manual III. Startup Starting the Hardware 1. Turn on the Mercury Burner Power Supply (Figure 4, below) and allow it to warm up for at least 5 minutes. It will emit faint clicks before turning green. When the light is solid green, it is ready.

  • Page 8: Starting The Software

    ICELL8® Imaging System User Manual Starting the Software 1. Turn on the computer and launch the Micro-Manager software: NOTE: Wait until all hardware is on and ready before starting the software, otherwise the software will generate an error message. 2. Once the Micro-Manager Copyright window opens, click OK to continue to the Main window.

  • Page 9: Operation

    Figure 10. Main window and Multi-Dimensional Acquisition window. The Multi-Dimensional Acquisition window acquires stacks of multiple images using various parameters. For single-cell images of ICELL8 chip nanowells, the software uses the “Multiple positions (XY)” setting to acquire images of 6 x 6 well regions.

  • Page 10: Figure 11. Live Window With Image Of 6 X 6 Well Region

    ICELL8® Imaging System User Manual NOTE: The wells will appear blurry as you focus below the top of the well to find the cells near the bottom of the well. Figure 11. Live window with image of 6 x 6 well region.

  • Page 11: Figure 12. Multi-Dimensional Acquisition Window

    ICELL8® Imaging System User Manual Figure 12. Multi-Dimensional Acquisition window. 7. Place the chip on the microscope Holding Platform with the chamfered (notched) corner facing the upper right corner (see Figure 13, below). Ensure that the chip is sitting flat on the chuck from all sides and is in the correct orientation.

  • Page 12: Focusing On Cells

    ICELL8® Imaging System User Manual 8. Remove the second release liner. Hold down the tab of the Imaging Film while peeling off the top layer release liner of the Imaging Film using tweezers. Save the peeled release liner. The chip will remain sealed by the double-sided intermediate film layer.

  • Page 13: Figure 17. Adjusting Camera Depth

    ICELL8® Imaging System User Manual 3. In the lower portion of the Main window, select the Contrast tab and select a value in the range of 12–14 bits in the Camera Depth menu. Figure 17. Adjusting camera depth. 4. Adjust the contrast of the chip image by selecting Autostretch and Ignore and a value in the range of 1–3% for Ignore (see Figure 18, below).

  • Page 14: Figure 19. Adjusting Microscope Focus

    ICELL8® Imaging System User Manual 5. Use the microscope manual focus (under the stage on both sides; see Figure 19, below) to achieve the optimal Z-plane focal depth to visualize a field of single cells. Figure 19. Adjusting microscope focus.

  • Page 15: Figure 21. Moving The Motorized Stage To Position 143

    ICELL8® Imaging System User Manual Figure 21. Moving the Motorized Stage to Position 143. 7. In the Main window, under “Configuration settings”, select Texas Red as the illumination channel for examination of fiducial-containing wells. Figure 22. Setting the illumination channel to Texas Red.

  • Page 16: Figure 24. Checking Cell-Stain Channels

    ICELL8® Imaging System User Manual 9. Select Pos143 in the Stage Position List window and confirm that all 36 nanowells are properly captured in the image. There will be four fiducial-containing nanowells in the shape of a square, as shown in Figure 23 (above). Position 143 is located at the lower right corner of the stage fixture.

  • Page 17: Acquiring Images

    ICELL8® Imaging System User Manual Acquiring Images 1. Click Aquire! and accept the prompt to create a new folder based on the current date. The camera will take images of the fields of view with both the DAPI and Texas Red filters. The entire image folder should contain 288 TIFF files from 144 fields of view (Pos0–Pos143);...

  • Page 18: Appendix A: Maintenance And Troubleshooting

    © 2017 Takara Bio Inc. All Rights Reserved. All trademarks are the property of Takara Bio Inc. or its affiliate(s) in the U.S. and/or other countries or their respective owners. Certain trademarks may not be registered in all jurisdictions. Additional product, intellectual property, and restricted use information is available at takarabio.com.

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