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logos biosystems CELENA X User Manual

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USER MANUAL
LBSM-MD-ML-CLX-001
www.logosbio.com
Rev. 5

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Summary of Contents for logos biosystems CELENA X

  • Page 1 USER MANUAL LBSM-MD-ML-CLX-001 www.logosbio.com Rev. 5...
  • Page 2 LED Filter Cubes are LTC Licensed Products provided under an intellectual property license from Life Technologies Corporation. © 2020 Aligned Genetics, Inc. All rights reserved. Logos Biosystems is the brand name of the life science business of Aligned Genetics, Inc.

  • Page 3: Table Of Contents

    Table of Contents GENERAL ................................... 4 1.1 A BOUT THIS USER MANUAL 1.2 E XPLANATION OF SYMBOLS SAFETY INFORMATION ............................5 2.1 D EVICE SYMBOLS 2.2 D EVICE SAFETY ERSONAL SAFETY AFETY STANDARDS GETTING STARTED ............................... 8 3.1 P RODUCT CONTENTS CELENA®...
  • Page 4 5.3 C HANGE OBJECTIVES 6.4 A DJUST OBJECTIVE CORRECTION COLLARS 6.5 V ERSION UPGRADE APPENDIX A: TROUBLESHOOTING.......................... 48 MAGE QUALITY XPLORER ECHANICAL APPENDIX B: SPECIFICATIONS ..........................50 CELENA® X H ONTENT MAGING YSTEM APPENDIX C: ORDERING INFORMATION....................... 51 NSTRUMENTS BJECTIVES FILTER CUBES CCESSORIES APPENDIX D: PURCHASER NOTIFICATION ......................

  • Page 5: General

    1. General 1.1 About this user manual This user manual provides the information necessary to operate the CELENA® X High Content Imaging System. PLEASE READ THIS USER MANUAL BEFORE OPERATING THE SYSTEM. If any part of this user manual is not clear, please contact Customer Support for assistance.

  • Page 6: Safety Information

    2. Safety information 2.1 Device symbols Electrical symbols Symbol Description Power symbol Safety symbol Symbol Description Risk group 3. UV emitted from this device may be hazardous. Avoid eye exposure. Invisible laser radiation. Avoid exposure to beam. Class 3b laser product.

  • Page 7: Device Safety

    Leave sufficient space around the device for air circulation and cooling. Take care that the device does not overheat during long and continuous operation. Use components provided or authorized by Logos Biosystems. If the proper combination of components is not used, product safety cannot be guaranteed.

  • Page 8: Personal Safety

    Community provisions for which this marking is required. Users must be aware of and follow the conditions described in this manual for operating the instrument. The protection provided by the device may be impaired if the device is used in a manner not specified by Logos Biosystems. Korean standards Symbol Description The KC certification mark indicates that this device conforms with Korea’s...

  • Page 9: Getting Started

    3. Getting Started 3.1 Product contents Your CELENA® X High Content Imaging System is shipped with the following components: CELENA® X main unit  Camera module (installed as ordered)  Condenser (installed as ordered)  Filter cubes (installed as ordered) ...
  • Page 10 CELENA® X Main unit CELENA® X Controller Accessories Single Slide Holder Microplate Holder Universal Vessel Holder Cable PS-1...
  • Page 11 Cable PS-2 Cable SIG Cable USB 2.0 Cable USB 3.0 (Optional) Cable Laser AF Power cord...

  • Page 12: Celena® Xmain Unit

    Flathead Screwdriver Product description CELENA® X CELENA® X is an integrated imaging system designed for rapid, high content image acquisition. Customizable imaging protocols, image-based and laser autofocusing modules, and motorized XYZ stage simplify slide scanning and well plate imaging. CELENA® X main unit CELENA®...

  • Page 13: Celena® X Controller

    CELENA® X cable connections Back of the CELENA® X main unit ① Laser AF: Deliver the output signal from the laser autofocus module. ② SIG: Transmit the communication signals between the main unit and the controller. ③ PS-1: Deliver DC power from the controller to the main unit. ④...

  • Page 14: Software

    CELENA® X Cell Analyzer is used to process and analyze images to quantify numerous cellular phenotypes simultaneously. CELENA® X Cell Analyzer also provides tools to edit and annotate images as well as create videos. CELENA X Cell Analyzer CELENA® X Labels...

  • Page 15: Setting Up

    CELENA® X Labels Laser safety label UV light safety label Top of the CELENA® X Main unit CELENA® X main unit label Back of the CELENA® X main unit CELENA® X Controller label Back of the CELENA® X Controller 3.3 Setting up...

  • Page 16: Unpacking

    Unpacking Move the unpacked boxes to the site of operation. CELENA® X main unit Open the CELENA® X box and remove the Styrofoam top and sides. Lift the CELENA® X out of its box by grasping its base firmly. Place the CELENA® X on a flat, level surface that is free of vibration. Anti-vibration tables are recommended for optimal use.

  • Page 17: Turn On The Celena® X

    Remove shipping restraints Unscrew screw A and pull it up to remove it from the LED filter cube stage cover. Unscrew screw B and pull it up to remove it from the X/Y stage. The shipping restraints must be removed before the CELENA® X is turned on. Store the screws in the accessories box for future use.

  • Page 18: Install Filter Cubes And Objectives

    Using other programs as well as CELENA® X Explorer at the same time can affect both imaging and analysis time. Using only CELENA® X Explorer is recommended when imaging and acquisition. Install filter cubes and objectives CELENA® X Explorer must be on to install filter cubes and objectives. For detailed instructions on how to install filter cubes, go to 5.2 Change filter cubes.

  • Page 19: Celena® X Explorer

    4. CELENA® X Explorer 4.1 User interface The CELENA® X Explorer is the graphical user interface for the CELENA® X High Content Imaging System. CELENA® X Explorer ① CHANNELS: Gives control over light, camera, and focus settings. ② VESSEL: Allows you to select the appropriate vessel, wells, and fields to capture. ③...

  • Page 20: Channels

    Channels This panel is used to set light, camera, and focus parameters for a project. ① Add: Adds the CHANNELS settings to the project protocol. ② Objectives: Allows you to select from the currently installed objectives. ③ Channels: Allows you to select from the currently installed filter cubes and adjust light and camera settings.
  • Page 21 In case of brightfield condenser, click the condenser (BF) button for transmitted light. The dropdown menu allows you to control the condenser’s iris diaphragm.  AUTO Automatically adjusts to accommodate the selected objective  100% Used for objectives with high magnification ...
  • Page 22 ① Focus slider: Used to adjust focus. The focus slider represents the full focal range. Adjust focus by moving the slider in the desired direction. ② Z-stage speed: Used to adjust the speed at which the Z-stage moves with each action.
  • Page 23 Multi-well plates at least 1 field/well ±100 µm Slides every 10-20 fields ±10 µm *Optimal AF frequency is also affected by objective magnification. Adjust accordingly. *Optimal AF range is also dependent on vessel bottom flatness. Adjust accordingly. When setting up Multiscan AF, you can select to use either the image-based or laser autofocus (optional;...

  • Page 24: Vessel

    Tips:  Pre-scan automatically adjusts optimal parameters in most cases. Perform Pre-scan at first.  Perform Test AF after Pre-scan to verify the laser autofocus properly finds a focal plane. If there is a slight difference from the best focal plane adjust Focus offset and run Test AF again.

  • Page 25: Z-Stack

    The acquisition order is initially set to start from Z-stack > Channel > Field > Well > Time lapse. If you want to have it start from Channel, please contact your local distributor or Logos Biosystems. Select Stitch: Click Stitch button if you want to acquire images with overlap. If the button is turned on the window to set Overlap Region will display.

  • Page 26: Time Lapse

    Start/End Start Well 2 Well 1 Well 3 Range (current) Well 2 Current position Well 1 Well 3 Z-stack with autofocus Well 2 AF position Well 1 Well 3 Z-stack methods  Start/End: Set the start and end positions of the Z-stack. Set start: Use to set the current focal plane as the start position.

  • Page 27: Project

     Maximum frame rate: Can only be used when imaging a single field in a single focal plane with one channel to capture up to 30 frames per second. This option can be used for high-speed experiments such as calcium imaging. Note: The interval will take into account other protocol settings such as the autofocus settings and exposure time.

  • Page 28: Toolbar

    ② Project details: Shows you the file path and name of a created project. ③ Open protocol: Allows you to open a previously saved protocol. This opens a previously saved protocol file (.cxprotocol). When you open a protocol, make the appropriate adjustments to each parameter as needed.

  • Page 29: Settings

    Settings Settings allows you to set system options and perform calibration procedures. To access the Settings window, click the Settings wheel above the tool bar. Camera The camera settings allows you to select the camera, bit depth, saturated pixel color, as well as auto white balance of the color camera.
  • Page 30 ① Change objectives: Can be used to install and remove objectives. ② Adjust correction collars: Can be used to adjust the correction collar of objectives. ③ Objective information: Can be used to set the installed objectives and label them. Filter cubes The filter cubes settings allows you to change filter cubes and set the pseudocolor and description for each installed filter cube.

  • Page 31: Workflow

    Vessels The vessels settings allows you to create and edit custom vessels. ① Vessel details: Can be used to select the vessel type, number of wells, well shape, and vessel name. ② Vessel description: Shows the associated catalog numbers of the selected vessel. ③...
  • Page 32 Overview Upon starting Explorer, you will create a new project protocol to capture images. Select vessel. ▼ Select objective. ▼ Select a channel. ▼ Set light and camera parameters. ▼ Focus on the sample. ▼ Adjust light. ▼ Set Multiscan AF options. ▼...
  • Page 33 Click the correct vessel in Product tab. It is crucial that you select the correct vessel to ensure proper focusing and vessel navigation. If the vessel you need is not available, go to Settings > Vessels to create a vessel. Be careful not to drop the vessel into the CELENA®...
  • Page 34 This device uses Class 3B laser and ultraviolet LEDs. Make the CELENA® X door is closed when imaging to protect your eyes. Direct exposure to and diffuse reflections of the laser can be hazardous to the eye. Minimize the time that the sample is being exposed to light to prevent from photobleaching and/or phototoxicity.
  • Page 35 Focus sharply. Move the focus slider in the desired direction or enter the desired value in the Z-position box. Alternatively, click Find Focus to have the CELENA® X find the optimal focal plane. Set the range to scan from the current focal position. A long search range is useful when finding the focal plane of an unknown object.
  • Page 36  Image-based: Make sure the current field is focused sharply. Set the range to scan from the current focal position. You can choose to turn the user-defined focus offset on or off. The user-defined focus offset means that the system will calculate the difference between what the system defines as the optimal focal plane and what the user defines as the focal plane of interest and automatically calibrate the focus accordingly.

  • Page 37: Save A Project Protocol

    Repeat for all necessary channels. 4. Select areas and fields to be In the VESSEL panel, select the well(s) to image in the vessel map. captured Click individual wells or drag and drop to select multiple wells. Wells selected for imaging will be filled with yellow.

  • Page 38: Load A Project Protocol

    Name the protocol and designate the file path. Load a project protocol To load a previously saved protocol, click Open Protocol in the PROJECT panel. Make the appropriate adjustments to each parameter as needed. This is especially important for the Multiscan AF feature and Z-stack imaging. Make sure to adjust Multiscan AF settings for each channel being imaged.

  • Page 39: Maintenance

    If liquid is spilled on the instrument, turn off the power and wipe dry immediately. Do not exchange components between instruments unless they have been provided or authorized by Logos Biosystems. Use only optical-grade cleaning materials to clean optical components.
  • Page 40 Click Next. Click the filter cube you want to change. The filter cube stage will move to that position. Unplug the connector (A) of the filter cube. Loosen the screw (B) in the cube with a flat-head screwdriver. Gently pull out the filter cube.
  • Page 41 Insert the desired LED filter cube, fasten the screw, and plug in its connector. Repeat as necessary. Click Finish when complete. This will return you to the original filter cubes settings window. ① Select the installed filter cube from the registered filter cubes list. ②...
  • Page 42 Click Start. Remove the vessel holder from the stage. Click Next. Click the objective you want to change. The turret will turn to that position.
  • Page 43 Grasp the objective at its base and unscrew it from the turret. Replace it with the desired objective and screw it in securely. If applicable, set the correction collar (A) as needed. Repeat as necessary. Click Finish when complete. This will return you to the original objectives settings window.
  • Page 44 ① Select the installed objective from the compatible objectives list. ② Select the post in which it was installed from the installed objectives list. ③ Click >>. ④ Double-click the label box to change how it shows up in the CHANNELS panel. If it is blank, the label will be the name of the objective.
  • Page 45 Click Next. Click the desired objective. The turret will turn to that position. Grasp the objective at its base and unscrew it from the turret. Set the correction collar (A) as needed.
  • Page 46 Reinstall the objective with care. Repeat as necessary. Click Finish to complete.
  • Page 47 Procedure Backup the files, CELENA X Explorer.ini and CELENA X_User.db to the desktop. You can find the CELENA X Explorer.ini in the Explorer folder where the executable file (exe.) is located and CELENA X_User.db file in the User folder. LogosBiosystems > CELENA X > CELENA X Exploer.ini LogosBiosystems >...
  • Page 48 After the uninstallation, check if the Explorer folder remains. (Logosbiosystems > Logosbiosystems > CELENA X > Explorer) If the Explorer folder exists, delete the folder. Install the new software. Overwrite the backup files of CELENA X Explorer.ini and CELENA X_User.db to the original locations.
  • Page 49 Appendix A: Troubleshooting Image quality Uneven focus Make sure the vessel bottom is clean and free of fingerprints. Place the vessel in the appropriate vessel holder. Make sure it fits snugly and lies flat. Make sure you focus sharply on a sample before setting up the autofocus for Multiscan AF. Make sure you have selected the correct vessel.
  • Page 50 Abnormal program termination Close the message window. Turn off the instrument. Check whether the cables of PS-1, PS-2 and SIG are properly connected. Reconnect USB2.0 and USB3.0 cables. Turn on the instrument. Restart Explorer If this window still appears contact your local distributor or the service engineer. Image irresponsive to changes Turn on the light in the CHANNELS panel.
  • Page 51 Pass, Cy5 Long Pass and custom filters Objective turret Motorized; 5 interchangeable objectives Compatible objectives 1.25-100X; Olympus, Zeiss, and Logos Biosystems objectives Only objectives with RMS-thread and the parfocal length of 60 mm are compatible. Condenser Motorized; basic or phase contrast condenser...
  • Page 52 Appendix C: Ordering Information Instruments Cat # Product CX30000 CELENA® X High Content Imaging System CELENA® X main unit  CELENA® X Controller  External PC with CELENA® X Explorer installed  Universal Vessel Holder  Microplate Holder  Single Slide Holder ...
  • Page 53 Cy5 Long Pass 620/60 665lp I10141 Custom I10142 Accessories Cat # Product I10411 Microscope Calibration Slide #1...
  • Page 54 Appendix D: Purchaser Notification Limited use label license Research use only The purchaser of this product should use this product only for research for the sole benefit of the purchaser. By use of this product, the purchaser agrees to be bounded by the terms of this limited use statement whether the purchaser is a for-profit or a not-for-profit entity.
  • Page 55 Logos Biosystems Aligned Genetics, Inc. HEADQUARTERS FL 2 & 3 28 Simindaero 327beon-gil, Dongan-gu Anyang-si, Gyeonggi-do 14055 OUTH OREA Tel: +82 31 478 4185 Fax: +82 31 360 4277 Email: info@logosbio.com 7700 Little River Turnpike STE 207 Annandale, VA 22003...

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