Nexcelom Bioscience Cellometer Auto T4 User Manual
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Nexcelom Bioscience Cellometer Auto T4 User Manual

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Summary of Contents for Nexcelom Bioscience Cellometer Auto T4

  • Page 2 Cellometer® is a registered trademark of Nexcelom Bioscience LLC. Other products referred to in this document may be either trademarks and/or registered trademarks of their...

  • Page 3: Table Of Contents

    Table of Contents CHAPTER 1. INTRODUCTION ....................1 Auto T4 System Overview........................1 Contents of Shipping Container ......................2 About this User Manual .......................... 3 Glossary of Abbreviations ........................4 Glossary of Symbols..........................5 CHAPTER 2. EQUIPMENT SAFETY .................... 7 Potential Hazards ...........................
  • Page 4 Table of Contents Cell Type Drop-down Defaults........................26 Importing Cell Types ............................. 26 From Nexcelom Cell Library ..............................26 From Custom Libraries ................................27 Deleting Cell Types............................27 From Home Screen................................27 From Import/Export Cell Type Screen ..........................28 Clearing Cell Types from Drop-down ......................28 Exporting Cell Types to a Library ........................
  • Page 5 Table of Contents Automatically Saving Data ..........................52 Automatically Saving Images ......................... 52 Loading Data and Images ........................53 Loading Data Files ............................53 Loading Image Files ............................53 Exporting Data and Images ........................54 CHAPTER 11. CLEANING, MAINTENANCE AND STORAGE ............55 Cleaning ..............................
  • Page 6 Table of Contents 8001009 Cellometer® Auto T4 User Manual Rev C...

  • Page 7: Chapter 1. Introduction

    Using Brightfield imaging and pattern-recognition software, the Cellometer Auto T4 quickly identifies individual cells and accurately calculates Cell Count, Mean Diameter (micron) and Cell Concentration (cells/mL). Customizable cell type parameters are designed to assist with the declustering of clumpy cell lines.

  • Page 8: Contents Of Shipping Container

    Note: Store USB drive where it can easily be located if software needs to be installed on another computer. Cellometer Auto T4 User Manual and Quick Start Guide Available as .pdf files on the USB Drive with software Figure 3. Chambered Slide...

  • Page 9: About This User Manual

    Chapter 1. Introduction ABOUT THIS USER MANUAL This Cellometer Auto T4 User Manual provides information on the following topics: System Overview and Shipping Contents Basic Counting and Analysis Workflow • • Potential Hazards and Safety Information Cell Types and Adjustment Parameters •...

  • Page 10: Glossary Of Abbreviations

    Chapter 1. Introduction GLOSSARY OF ABBREVIATIONS The following abbreviations may be displayed on the shipping container, on the Cellometer Auto T4 device label or in this user manual. Amperes Kilowatt Alternating Current Pounds American National Standards Institute Personal Computer ANSI...

  • Page 11: Glossary Of Symbols

    Chapter 1. Introduction GLOSSARY OF SYMBOLS The following international symbols may be displayed on the shipping container, on the Cellometer Auto T4 device label or in this user manual. Fragile, Handle with Care Keep Dry ISO0626 per This End Up ISO0623 per ISO0621 per EN ISO 15223-1.
  • Page 12 Chapter 1. Introduction 8001009 Cellometer® Auto T4 User Manual Rev C...

  • Page 13: Chapter 2. Equipment Safety

    Chapter 2. Equipment Safety As with any equipment that involves moving parts, there are potential hazards involved in the operation and maintenance of this instrument. This chapter identifies potential hazards and suggests precautions to avoid them. POTENTIAL HAZARDS This section describes instrument safety features designed to minimize potential hazards. Before using the system, familiarize yourself with this information.
  • Page 14 Chapter 2. Equipment Safety Instrument System Hazard Read the instructions, warnings and cautions provided with the instrument before using. WARNING: Inspect instruments and cables for breaks, cracks, nicks and other damage before every use. This may be done visually under magnification or with a high voltage insulation testing device. If damaged, do not use.

  • Page 15: Safety Information

    Chapter 2. Equipment Safety SAFETY INFORMATION Personnel operating and maintaining the Auto T4 instrument should be familiar with the safety information included in this section. Safety Protocols Nexcelom assumes no liability whatsoever for any damage, loss or injury resulting from an application of a product that is not in strict accordance with the instructions provided with the product.
  • Page 16 Chapter 2. Equipment Safety 8001009 Cellometer® Auto T4 User Manual Rev C...

  • Page 17: Chapter 3. Instrument Description

    Chapter 3. Instrument Description This chapter describes the Auto T4 instrument components, device label and system specifications. AUTO T4 COMPONENTS Front View Back View Device Label Auto Sample Slot Focus Knob Auto Sample Slot: Chambered slides are inserted through this slot for viewing (accepts only one slide at a time). Focus Knob: Radial dial used to focus the camera.

  • Page 18: System Specifications

    The Auto T4 System is comprised of the instrument connected via USB cable to an Operating Computer used to run the instrument software. The Operating Computer can be linked to a network for accessing external files, printers and storing count results. Cellometer Auto T4 Cell Counter Channels Brightfield...

  • Page 19: Chapter 4. Unpacking And Site Preparation

    Nexcelom Bioscience has designed an Installation Qualification/Operation Qualification (IQ/OQ) validation specifically for the Cellometer Auto T4. Our experienced Support team is available to assist your organization during set up of the instrument and while performing the IQ/OQ validation. Contact Nexcelom Support for more information at +1 (978) 327-5340 or via email: support@nexcelom.com...

  • Page 20: Site Preparation

    Chapter 4. Unpacking and Site Preparation SITE PREPARATION Facility Requirements Instrument must be plugged directly into a surge protector (recommended) or power outlet. Ensure all cables are free from kinks or tangles prior to starting the Auto T4. Due to how the instrument is used with liquids, the instrument should be placed on a level surface. Keep the area around the Auto T4 clean between, during and post operation.

  • Page 21: Chapter 5. Operation

    Chapter 5. Operation This chapter describes installing the software and steps to be performed when using instrument for the first time. WARNING: Never force the door open or place a foreign object inside the Auto Sample Slot. Should anything get stuck or spill inside the sample slot, power OFF the instrument and contact Nexcelom Support for service at +1 (978) 327-5340 or via email: support@nexcelom.com INSTALLING THE SOFTWARE...
  • Page 22 5. Click Install to install the instrument software on the hard drive of the Operating Computer. A screen containing prerequisite software to be installed on your computer may be displayed. Click Install to continue. 6. When prompted, click OK to check for prior versions of Cellometer Auto T4 software. 7. Click Finish to exit the installer.

  • Page 23: Using Instrument For The First Time

    Chapter 5. Operation USING INSTRUMENT FOR THE FIRST TIME Starting the Software 1. On the desktop of Operating Computer, launch the Auto T4 software by double-clicking the Cellometer Auto software icon. 2. Enter instrument Serial Number from the device label into the input fields. The device label can be found on back of the instrument.

  • Page 24: Taking A Background Image

    Chapter 5. Operation Taking a Background Image After initial setup of the instrument, you must take a background image of the system by itself (without the cell counting chambered slide inserted) to normalize the cell counter. Unless the instrument is moved to a new location, there is generally no need to take another background image.
  • Page 25 Chapter 5. Operation 6. Once the image is complete, a pop-up message indicates the background image has been saved. Click OK. 7. Click the Display Image button to view the background image taken. 8. Confirm that the background is a uniform gray color. If there is any discoloration (e.g., light/dark areas) of the image, contact Nexcelom Support for assistance at +1 (978) 327-5340 or via email: support@nexcelom.com...
  • Page 26 Chapter 5. Operation 8001009 Cellometer® Auto T4 User Manual Rev C...

  • Page 27: Chapter 6. Software Gui Elements

    Chapter 6. Software GUI Elements This chapter presents key graphical user interface (GUI) menus and screens available in the Auto T4 software. Note: The Auto T4 software used to run the instrument must be installed on the Operating Computer before you can use the GUI.
  • Page 28 Chapter 6. Software GUI Elements Sample ID – Allows entry of a unique sample identifier. Dilution Factor – Allows entry of final dilution factor for cell sample to calculate accurate concentration. Live/Dead Count – Displays numbers of live and dead cells found. Displayed only if Test Viability is selected.

  • Page 29: Available Menus

    Chapter 6. Software GUI Elements AVAILABLE MENUS The following menus are available in the Menu Bar displayed across the top of the Home screen. File Menu Load a previously saved image and display it. Load a previously saved image and count it. Choose a new data file.

  • Page 30: Help Menu

    Frequently Asked Questions/Application Notes (an internet connection is required when using this option). Displays version number of the Cellometer Auto T4 software currently installed. Note: Alternatively, clicking the Help icon located in the bottom right corner of the Home screen also displays online help contents and resources, a link for submitting a Support ticket, and Nexcelom contact information.

  • Page 31: Import/Export Cell Type Screen

    Chapter 6. Software GUI Elements IMPORT/EXPORT CELL TYPE SCREEN Cell Types define image analysis parameters used in cell sample detection. The Import/Export Cell Type Screen is used to import either standard cell types from the Nexcelom Cell Library installed with the Auto T4 software or custom cell types from an external network location.

  • Page 32: Cell Type Drop-Down Defaults

    Chapter 6. Software GUI Elements Cell Type Drop-down Defaults The Auto T4 displays several commonly known cell types that have been pre-selected to appear in the Cell Type drop-down menu available on the Home screen as initial defaults. These cell types are listed below: •...

  • Page 33: From Custom Libraries

    Chapter 6. Software GUI Elements FROM CUSTOM LIBRARIES 1. In the Import from Library section of the Import/ Export Cell Type screen, click the custom cell library radio button to choose a custom library (if available) or click Browse to navigate to folder containing a custom library and double-click on a library name to select it.

  • Page 34: From Import/Export Cell Type Screen

    Chapter 6. Software GUI Elements FROM IMPORT/EXPORT CELL TYPE SCREEN 1. From the Home screen, open the Cell Type Menu in the menu bar and select the Import/Export… option. 2. In the Cell Types in Drop-Down Menu pane (displayed on the right), highlight the cell type to be deleted. 3.

  • Page 35: Exporting Cell Types To A Library

    Chapter 6. Software GUI Elements Exporting Cell Types to a Library Users can export cell types selected to appear in the Cell Type drop-down menu available on the Main Screen (displayed in the right pane) to create a custom Cell Library. 1.
  • Page 36 Chapter 6. Software GUI Elements 8001009 Cellometer® Auto T4 User Manual Rev C...

  • Page 37: Chapter 7. Sample Preparation

    Chapter 7. Sample Preparation PREPARING CELLOMETER CHAMBERED SLIDES 1. For SD100 slides, remove the protective film from both sides of the slide. PD100 slides do not have this film. Note: It may be difficult to peel film from the slide. Some clients use tape to grab the film and then pull it off. Do not touch or write on the clear optical window areas of the slide.

  • Page 38: Loading Sample Into Counting Chamber

    Chapter 7. Sample Preparation LOADING SAMPLE INTO COUNTING CHAMBER Cellometer chambered slides consist of two independent counting chambers manufactured to a precisely- controlled volume. A cell suspension of 20 µL is pipetted into a chamber through one of the induction ports. 1.

  • Page 39: Chapter 8. Counting And Analysis Workflow

    Chapter 8. Counting and Analysis Workflow This chapter presents basic workflow for counting cells/beads using the Auto T4, including best practices and tips. Sample images are provided as a reference for many common cell types. PERFORM CELL COUNTING 1. Power ON the instrument and from the desktop of the Operating Computer, launch the Auto T4 software. 2.
  • Page 40 Chapter 8. Counting and Analysis Workflow 5. Slowly turn the Focus Knob on side of the instrument until good focus is achieved to ensure accurate counts. Live cells will have a bright center and a dark, crisp clearly defined edge as shown in the examples below. 6.

  • Page 41: Review Count Results

    Chapter 8. Counting and Analysis Workflow REVIEW COUNT RESULTS Once counting is complete, count results are displayed in the bottom left and right corners of the screen. Use the controls in the View Image area to view different areas of the counting chamber. Confirm data is accurate by clicking the Counted check box to view image using a graphic overlay outlining counted cells (e.g., green for counted cells and yellow for cells not counted because they were larger than the specified cell diameter).

  • Page 42: Best Practices And Workflow Tips

    Chapter 8. Counting and Analysis Workflow Best Practices and Workflow Tips The progress bar at the bottom of the software panel indicates counting is in progress. A numeric counter also • displays updated cell counts. • Count results displayed in the bottom left corner of the screen represent the entire counting chamber and include Cell Count, Mean Diameter (micron) and Cell Concentration (cells/mL).

  • Page 43: View Cell Size Analysis

    Chapter 8. Counting and Analysis Workflow VIEW CELL SIZE ANALYSIS The Cell Size Analysis tool displays cell size data from count results by generating a histogram that enables users to optimize cell diameter parameters for the sample (e.g., to exclude debris or very large cells) and refine count data. By overlaying data from multiple samples into a single histogram, users can create an analysis of the change in cell diameter size over time.

  • Page 44: Calculate Sample Adjustment

    Chapter 8. Counting and Analysis Workflow CALCULATE SAMPLE ADJUSTMENT The Sample Adjustment tool assists you in calculating the adjustment necessary in order to achieve a specified target concentration or target number of cells in a tube. Before calculating this adjustment, you must first perform a count on either a live image or saved image to display the current cell count and concentration (or live/dead count, total cell count, viability %, live cell concentration and total cell concentration if Test Viability was selected).

  • Page 45: Reference Of Cell Sample Images

    Common cell type and bead sample images taken by the Auto T4 are presented for your reference below. Visit the Cellometer Auto T4 page on our website, click the Applications tab and scroll down to find additional cell type sample images.
  • Page 46 Chapter 8. Counting and Analysis Workflow 8001009 Cellometer® Auto T4 User Manual Rev C...

  • Page 47: Chapter 9. Cell Types

    Chapter 9. Cell Types Chapter 9. Cell Types This chapter describes editing customizable cell type parameters to assist with declustering of clumpy cell lines. In addition, it lists all cell types included in the built-in Nexcelom Cell Library. EDITING CELL TYPES Cell types can be modified to meet your requirements using parameters available on the Cell Type Screen.
  • Page 48 Chapter 9. Cell Types Maximum – Indicates maximum cell diameter to be counted. Roundness – Indicates minimum roundness factor to be counted. Contrast Enhancement – Defines contrast enhancement for cells in relation to the background. Default Dilution Factor – Indicates if a default dilution factor should be used and enables entry of a specific value.

  • Page 49: Adjusting Cell Diameter Parameters

    Chapter 9. Cell Types Adjusting Cell Diameter Parameters For each cell type, Cell Diameter Minimum/Maximum (in microns) parameter settings indicate the size of cells to be included in the count To determine if cell diameter parameters are set up properly, use the Counted check box in the Home screen to display a graphical overlay outlining counted cells (e.g., green for counted cells and yellow for cells not counted as they are larger than the specified cell diameter).

  • Page 50: Adjusting Trypan Blue Viability Parameters

    Chapter 9. Cell Types Adjusting Trypan Blue Viability Parameters Using default settings for Trypan Blue Viability Parameters provides the fastest counting speed for standard cell types stained a solid dark blue by trypan blue. As some cell types exhibit different morphology after being stained, it may be necessary to adjust parameters for accurate detection of the trypan blue population as indicated below.

  • Page 51: Adjusting Decluster Parameters For Clumpy Cells

    Chapter 9. Cell Types CHO cell count results were analyzed using different "Sensitivity" settings. Figure (a) shows CHO cell image while figures (b) – (e) show an incremental increase in “Sensitivity” parameter from 1 to 4, respectively. Green outlines indicate live cells while red outlines indicate dead cells.

  • Page 52: Sample Clumpy Mcf7 Cells

    Chapter 9. Cell Types SAMPLE CLUMPY MCF7 CELLS In the sample below, clumpy MCF7 cells are analyzed using both default and modified Decluster Edge Factor and Decluster Th Factor parameter settings. Edge Factor = 0.5 Edge Factor = 0.7 Th Factor = 1.0 Th Factor = 0.8 MCF7 cell count results were analyzed using different "Decluster"...
  • Page 53 Chapter 9. Cell Types Mouse thymocyte count results using the parameter settings identified in the screen above are shown in figure (a) with the Decluster feature activated (i.e., Do not Decluster Clumps check box is not selected) and in figure (b) without the Decluster feature.

  • Page 54: Creating New Cell Types

    Chapter 9. Cell Types CREATING NEW CELL TYPES Using Cell Type Wizard 1. Open the Cell Type menu from the menu bar and select the Cell Type Wizard… option. 2. Select the source of new cell parameters: Live Image or Saved Image 3.

  • Page 55: Nexcelom Cell Library

    Chapter 9. Cell Types NEXCELOM CELL LIBRARY The complete collection of cell types available for import from the Nexcelom Cell Library is listed below. 2372 DU145 HUVEC MDA-MB-435 Raji DU145 G37 Hybridoma MDA-MB-468 Ramos I9.2 MDBK 786-O Initial Cell Type medium size cells RCC45 A204...
  • Page 56 Chapter 9. Cell Types 8001009 Cellometer® Auto T4 User Manual Rev C...

  • Page 57: Chapter 10. Data Management

    Chapter 10. Data Management This chapter describes how to save count result data and image files, export them to an Excel spreadsheet and how to load previously-saved data and images back into the Auto T4 for reanalysis. SAVING DATA AND IMAGES Manually Saving Data 1.

  • Page 58: Automatically Saving Data

    Chapter 10. Data Management Automatically Saving Data 1. Open the Options Menu and select the Save Options… option. 2. Click the Auto Save data.txt check box to select it. 3. If you want to create a new data text file each time cells are counted, click the Create new data.txt with each sample check box to select it.

  • Page 59: Loading Data And Images

    Chapter 10. Data Management LOADING DATA AND IMAGES Loading Data Files 1. Open the File Menu and select the Open Data File option. 2. Navigate to a folder on the Operating Computer or network where data files are stored and select data file to be loaded.

  • Page 60: Exporting Data And Images

    Chapter 10. Data Management EXPORTING DATA AND IMAGES Data and images can be exported by clicking the Export to Excel icon after performing a cell count. 1. To Export Count Details to Excel, choose items to be included in the export (e.g., images with data file, JPG image copies with file, counted images with file) and indicate whether to open Excel when Done.

  • Page 61: Chapter 11. Cleaning, Maintenance And Storage

    Chapter 11. Cleaning, Maintenance and Storage Keeping the Auto T4 and its operative area clean between runs, during use and post runs is a best practice and prevents contamination. Caring for the instrument and its consumables is also a best practice. CAUTION: Always power OFF the instrument before cleaning as damage to the machine could occur.

  • Page 62: Storage

    Chapter 11. Cleaning, Maintenance and Storage Preventive Inspection and Maintenance Regular preventive inspections should be carried out to reduce safety concerns of the instrument due to aging, normal wear and tear, etc. The manufacturer assumes no responsibility for improper changes or repairs carried out on the instrument or its accessories by unauthorized persons.

  • Page 63: Chapter 12. Troubleshooting And Faqs

    Chapter 12. Troubleshooting and FAQs This chapter lists troubleshooting steps for resolving potential issues, common software error messages and Frequently Asked Questions (FAQs). TROUBLESHOOTING AND ERROR MESSAGES Particles appear in the background/background is too dark • Take a new background image by following the steps presented in Taking a Background Image on page 18.

  • Page 64: Frequently Asked Questions

    Chapter 12. Troubleshooting and FAQs IF ERROR REQUIRES CREATING A SUPPORT TICKET If error encountered that cannot be resolved and/or the problem persists after rebooting the Auto T4 system, perform the following steps before creating a Support ticket: 1. Record the error message. 2.

  • Page 65: Chapter 13. Nexcelom Support

    Lawrence, Massachusetts 01843 Phone: (978) 327-5340 Fax: (978) 327-5341 support@nexcelom.com San Diego Office Manchester Office Shanghai Office Nexcelom Bioscience LLC Nexcelom Bioscience Ltd Nexcelom Bioscience Instruments 11100 Roselle Street, Suite B Unit 5, Rutherford House (Shanghai) Co. Ltd San Diego, California 92121...
  • Page 66 Chapter 13. Nexcelom Support 8001009 Cellometer® Auto T4 User Manual Rev C...

  • Page 67: Appendix A. Consumables

    Appendix A. Consumables This appendix presents Nexcelom consumables designed specifically for the Auto T4 such as disposable chambered slides, beads and reagents. Catalog numbers are listed for all available sizes. DISPOSABLE CHAMBERED SLIDES Cellometer Cell Counting Chambered Slides are compatible with all Cellometer Systems. Each all-plastic, disposable slide contains two sample counting chambers with a precisely-controlled height.
  • Page 68 Packed with protective film. 5 Cases Good for 9,000 counts 1-year expiration. Visit the Nexcelom-products/cellometer-disposable-counting-chambers page on our website for an up-to-date list and to purchase Cellometer Counting Chambered slides directly from Nexcelom Bioscience. 8001009 Cellometer® Auto T4 User Manual Rev C...

  • Page 69: Beads And Reagents

    Appendix A. Consumables BEADS AND REAGENTS Nexcelom Bioscience provides a wide variety of beads and reagents for all of your analysis needs. Catalog # Description Size Unit Image 5 µm polystyrene bead in Trypan Blue B05-02-050 1 mL 4 Vials Solution.
  • Page 70 Appendix A. Consumables 8001009 Cellometer® Auto T4 User Manual Rev C...

  • Page 71: Appendix B. Declaration Of Conformity

    Address: 360 Merrimack Street, Suite 200, Lawrence MA 01843 USA Product Name: Cellometer Auto T4 High-Speed Cell Counter Model Number: Cellometer Auto T4 Conforms to the appropriate country standards and governing regulations listed below. We, as the manufacturer, are fully responsible for the design and production of the above-mentioned equipment.
  • Page 72 Appendix B. Declaration of Conformity 8001009 Cellometer® Auto T4 User Manual Rev C...

  • Page 73: Appendix C. Warranty And License Details

    (including unauthorized upgrades), disclose it to third parties or use it for any purpose other than running your Cellometer system. Nexcelom Bioscience LLC does not grant you any other rights to use or disclose the original Cellometer software or upgrades, and any further uses will be prosecuted by Nexcelom Bioscience LLC to the maximum extent possible by law.

  • Page 74: Limitation Of Liability (Hardware And Software)

    Nexcelom Proprietary Information without Nexcelom’s written consent. Nexcelom Bioscience LLC retains all ownership rights in the Nexcelom Proprietary Information and, other than limited license set forth in this section, Buyer shall have no right in or to the Nexcelom Proprietary Information.
  • Page 75 Cellometer® Auto T4 User Manual 8001009, Rev C 360 Merrimack St, Suite 200 Lawrence, MA 01843 USA T: +1 (978) 327-5340 F: +1 (978) 327-5341 info@nexcelom.com www.nexcelom.com...

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