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Nexcelom Bioscience Cellometer Auto T4 User Manual Download Page 23

 

Chapter 5. Operation 

8001009 Cellometer® Auto T4 User Manual Rev C 

 

17 

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. 

3.

 

Click 

Done 

to save the instrument Serial Number. 

4.

 

A pop-up message will appear indicating a background image needs to be taken before counting.  
Click 

OK 

to continue. 

 

 

 

Summary of Contents for Cellometer Auto T4

Page 1: ......

Page 2: ... Nexcelom assumes no responsibility for errors or omissions or for damages resulting in the use of information contained in this document or from the use of programs and source code that may accompany it In no event shall the publisher and author be liable for the loss of profit or any other commercial damage caused or alleged to have caused directly or indirectly by this document Cellometer is a ...

Page 3: ...3 INSTRUMENT DESCRIPTION 11 Auto T4 Components 11 Device Label 11 System Specifications 12 CHAPTER 4 UNPACKING AND SITE PREPARATION 13 Unpacking the Instrument 13 Site Preparation 14 CHAPTER 5 OPERATION 15 Installing the Software 15 Using Instrument for the First Time 17 Starting the Software 17 Taking a Background Image 18 CHAPTER 6 SOFTWARE GUI ELEMENTS 21 Home Screen 21 Available Menus 23 File ...

Page 4: ...rm Cell Counting 33 Review Count Results 35 Best Practices and Workflow Tips 36 Saving Data and Images 36 Printing Data 36 View Cell Size Analysis 37 Calculate Sample Adjustment 38 Reference of Cell Sample Images 39 CHAPTER 9 CELL TYPES 41 Editing Cell Types 41 Cell Type Screen 41 Adjusting Cell Diameter Parameters 43 Adjusting Trypan Blue Viability Parameters 44 Adjusting Decluster Parameters for...

Page 5: ...55 Routine Maintenance 55 Storage 56 CHAPTER 12 TROUBLESHOOTING AND FAQS 57 Troubleshooting and Error Messages 57 Frequently Asked Questions 58 CHAPTER 13 NEXCELOM SUPPORT 59 Scope of Support Services 59 Contacting Support 59 APPENDIX A CONSUMABLES 61 Disposable Chambered Slides 61 Beads and Reagents 63 APPENDIX B DECLARATION OF CONFORMITY 65 APPENDIX C WARRANTY AND LICENSE DETAILS 67 Warranty Inf...

Page 6: ...Table of Contents 8001009 Cellometer Auto T4 User Manual Rev C vi ...

Page 7: ...d and Total cell counts Viability and Live and Total cell concentrations including a helpful graphic overlay that may be used to highlight counted live and dead cells with color coded outlines in the image being viewed The small footprint of the Auto T4 is designed to fit in small lab spaces and Cellometer Auto software can be loaded on most computers With an Operating Computer connected to the in...

Page 8: ...slide contains two separate counting chambers Nexcelom provided Laptop Figure 4 If a laptop was purchased from Nexcelom optional along with the instrument it will be shipped in a separate box This laptop will have the specifications listed below Windows 10 OS 64 bit Intel Core Processor i3 8130U 2 20GHz 4GB RAM 500GB7200RPM SATA NoteBook Drive 15 6 1366 x768 HD LED Wide Screen If a laptop was not ...

Page 9: ... Software Installation and First time Use Troubleshooting and FAQs Key Software Screens and Menus Nexcelom Support Services and Contacts Slide Preparation and Loading Samples Consumables Conformity and Warranty The following Precaution Signifiers are used in conjunction with the symbol in this user manual IMPORTANT Note indicating that to skip or move past content_of_note may result in improper fu...

Page 10: ...de at MHz Megahertz BR Brightfield µL Microliter C Degrees Celsius µm Micron Cm Centimeter mL Milliliter EU European Union mm Millimeter FAS Field Application Scientist ms Millisecond FCC Federal Communications Commission nm Nanometer FDA Food and Drug Administration OQ Operational Qualification FL Fluorescence OSHA Occupational Safety and Health Administration GUI Graphical User Interface P N Par...

Page 11: ...d on shipping container FCC Part 15 Supplier Declaration of Conformity Located in user manual Declaration of Conformity to Medical Device Directive 93 42 EEC Located on SN label and in user manual ISO 9001 2015 Certified Follow Instructions for Use Located in user manual On Power Connection to Mains Located on instrument s Power Switch Off Power Disconnection from Mains Located on instrument s Pow...

Page 12: ...Chapter 1 Introduction 8001009 Cellometer Auto T4 User Manual Rev C 6 ...

Page 13: ...to fully understand and comply with local state and federal laws rules and regulations in the use of this system Cables and accessories not specified within the instructions for use with this instrument are not authorized Using other cables and or accessories may adversely impact safety and performance Note The Auto T4 instrument should not be used adjacent to or stacked with other equipment unles...

Page 14: ...ument CAUTION Do not stack equipment on top of the Instrument or place the Instrument on top of electrical equipment This is an unstable configuration and does not allow for adequate cooling In addition it is recommended that as much distance as possible be provided between the instrument and any equipment emitting a high vibration signature High levels of vibration may affect clarity of the image...

Page 15: ...nnection prior to use The Auto T4 must be set regulated and used in accordance with the instructions outlined in this user manual Failure to observe safety warnings and precautions may present a risk Only individuals with appropriate safety training and knowledge should operate assist in the operation of or perform cleaning and routine maintenance of this instrument Only the operator should be res...

Page 16: ...Chapter 2 Equipment Safety 8001009 Cellometer Auto T4 User Manual Rev C 10 ...

Page 17: ...to focus the camera Power Switch Controls power going to the instrument USB 2 0 Port Port used to connect the Auto T4 to the Operating Computer Power Cord Connector Input Port used to connect the power supply to the Auto T4 Device Label Lists the instrument serial number SN model manufacturer and input power requirement DEVICE LABEL Serial Number Manufacturer Polarity DC Power Connector Waste Elec...

Page 18: ...ults Cellometer Auto T4 Cell Counter Channels Brightfield Number of Channels 1 Commonly Used Compatible Dyes Trypan Blue Counting Speed Per Sample 30sec Volume per chamber 20 µL Cell Size Diameter Range 5 60 µm most mammalian cells Concentration Range 1x105 1x107 Software Cellometer Auto Pattern recognition software to quickly and accurately decluster identify and count individual cells Weight 10 ...

Page 19: ...en the outer box 4 Remove protective packaging 5 Remove the inner box 6 Open the secondary box to remove the Power Cord USB Computer Cable and Software USB 7 Remove the Auto T4 8 Place the Auto T4 in the prepared area see Facility Requirements on page 14 9 Remove the protective seals around the Auto T4 10 Insert the Power Cord Connector into the Auto T4 11 Plug the Power Cord into a surge protecto...

Page 20: ... position the device such so that it is difficult to disconnect from power main Transporting the Instrument Prior to transporting the instrument by vehicle disconnect the main from the wall and package all accessories and consumables individually and safely ensuring cables are also protectively wrapped Refer to instrument dimensions to ensure vehicle to be used for transporting meets these require...

Page 21: ...ong with the instrument the software was pre installed on the laptop and you can skip this section Continue to Using Instrument for the First Time on page 17 Note Ensure that instrument is not connected to the computer during software installation until specified To install the Cellometer software on a computer that was not purchased from Nexcelom follow these steps 1 Insert Software USB Drive tha...

Page 22: ...oftware 7 Click Finish to exit the installer 8 Confirm that Cellometer Auto software icon now appears on desktop of Operating Computer 9 Remove the Software USB and store it according to personal company guidelines 10 Plug power cord for the Auto T4 instrument into a surge protector recommended or outlet and power ON 11 Once software installation is complete connect USB 2 0 cable between the Auto ...

Page 23: ... 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 3 Click Done to save the instrument Serial Number 4 A pop up message will appear indicating a background image needs to be taken before counting Click OK to continue ...

Page 24: ...ter Unless the instrument is moved to a new location there is generally no need to take another background image 1 On the desktop of Operating Computer launch the Auto T4 software by double clicking the Cellometer Auto icon 2 Navigate to the Options Menu and select the Take Background Image option 3 Remove the chambered slide from the instrument if applicable 4 Click OK to confirm the Auto Sample ...

Page 25: ...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 9 Return to the Home screen to begin counting samples ...

Page 26: ...Chapter 5 Operation 8001009 Cellometer Auto T4 User Manual Rev C 20 ...

Page 27: ...r was purchased from Nexcelom along with the instrument the software was pre installed on the laptop and is ready for use HOME SCREEN The Home Screen also referred to as the main screen in the Auto T4 software is described below 1 Display Image Button Shows cells currently in the counting chamber Not enabled if a saved image has been loaded for display 2 Count Button Initiates counting procedure N...

Page 28: ...so users can indicate where count result data and images should be saved 15 Image Area Displays image captured by the instrument 16 Save Data Button Enables users to save the data in the indicated file 17 View Saved Data Button Displays the data that would be saved 18 Calculate Concentration Adjustment Icon Activates Sample Adjustment tool used to calculate volume needed to adjust concentration ca...

Page 29: ...7 Print results of the last count 8 Export results of the last count to MS Excel 9 Exit the software Cell Type Menu 1 Launch the Cell Type Wizard 2 Import Export cell types 3 Create a new cell type 4 Edit the currently selected cell type 5 Delete the currently selected cell type 6 Set the current settings as default Options Menu 1 Edit the counting options 2 Edit the save options 3 Take new backgr...

Page 30: ... a support ticket including sample images and access to Frequently Asked Questions Application Notes an internet connection is required when using this option 3 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 subm...

Page 31: ...e a new library 1 Selects Nexcelom Cell Library to display Nexcelom cell types in the Cell Types in Cell Library pane 2 Selects a custom library to display available cell types in the Cell Types in Cell Library pane If no library name is displayed click the Browse button and navigate to an external cell type library 3 List of cell types in the currently selected Cell Library 4 Imports the highligh...

Page 32: ...down as necessary from the Nexcelom Cell Library See Nexcelom Cell Library on page 49 for a complete list of available cell types Importing Cell Types Used to import cell types from the Nexcelom Cell Library or from external custom libraries Individual cell types can then be selected to appear in the Cell Types drop down menu displayed on the Home screen FROM NEXCELOM CELL LIBRARY The Nexcelom Cel...

Page 33: ... Click Import Highlighted to import highlighted cell type to appear in the Cell Types in Drop down Menu pane displayed on the right Note If the custom library selected is of a manageable size for the Cell Types drop down menu Import All can be used to import all cell types from the library Deleting Cell Types FROM HOME SCREEN 1 Use the Cell Type drop down menu to select the cell type to be deleted...

Page 34: ...essary If cell type was from a custom library or created using the Auto T4 it may be permanently deleted unless cell type was exported and saved prior to deletion Clearing Cell Types from Drop down 1 From the Home screen open the Cell Type Menu in the menu bar and select the Import Export option 2 To clear all cell types from the Cell Types in Drop down Menu pane displayed on the right select the ...

Page 35: ...e right pane to create a custom Cell Library 1 From the Home screen open the Cell Type Menu in the menu bar and select the Import Export option 2 To export cell types listed in the Cell Types in Drop down Menu pane displayed on the right select the Drop down Menu Options check box 3 Click the Export to Create Library button 4 Click OK to confirm save 5 Save file in designated folder 6 Click the Do...

Page 36: ...Chapter 6 Software GUI Elements 8001009 Cellometer Auto T4 User Manual Rev C 30 ...

Page 37: ...nd then pull it off Do not touch or write on the clear optical window areas of the slide 2 Place the Cellometer disposable chambered slide on a fresh Kimwipe 3 To prepare two samples at once label the individual chambers e g Chamber 1 and Chamber 2 in the white margin of each chamber Take care to ensure the clear portion of the counting chamber is not touched Note Both chambers of the slide do not...

Page 38: ...ep 1 above When preparing a sample to test for cell viability ensure that the trypan blue stock concentration used is 0 2 If stock concentration is 0 4 it is recommended you dilute it with a balanced salt buffer e g PBS and filter diluted solution with a 0 2 micron filter before adding stain to your sample 2 Immediately transfer 20 µL of sample into a counting chamber by pipetting directly onto sa...

Page 39: ...into counting chamber See Preparing Cellometer Chambered Slides on page 31 for details 3 Load the chambered slide into the Auto T4 by inserting the counting chamber that contains the cell sample into Auto Sample Slot until touching the internal stop The chamber inside the instrument will be counted Note If testing for cell viability you must stain the sample before loading it into the slide chambe...

Page 40: ...t center and a dark crisp clearly defined edge as shown in the examples below 6 In the Setup area of the screen choose the Cell Type from the drop down list enter a Sample ID and ensure the Dilution Factor is accurate for the sample 7 Click the Count button while the cell image is displayed Note When software is in Data Analysis Mode the Count button will not be enabled Click the Recount icon to r...

Page 41: ...for cells not counted because they were larger than the specified cell diameter Note When viewing counted cells a red line will appear along edges of the Viewing Pane identifying the outer perimeter of the counting area Any cells appearing on or outside of this red line are not counted If testing for cell viability select the Test Viability check box in the Setup area Additional count results incl...

Page 42: ...fying cell type parameters click the Recount Image icon to recount cells in the image Data analysis features include Sample Adjustment and Cell Size Analysis tools Based on the measured cell concentration you can calculate the sample adjustment to achieve a target concentration or cell number In addition cell size data for the sample can be displayed using a histogram See View Cell Size Analysis o...

Page 43: ...ots cell size diameter distribution Size Microns on the X axis against the Percentage of Total Count Percent on the Y axis with the scales of each axis determined by count results The coloring of bars reflects the nature of cells similar to how the graphic overlay is used to highlight counted cells e g if testing for viability green outlines live cells while red outlines dead cells Other features ...

Page 44: ...1 Click on the Sample Adjustment icon to launch the tool 2 Choose the concentration source to be used for calculating the adjustment live dead or total concentration and click Continue 3 Enter the Original Sample Volume The Total Cell Number in Sample value will be updated according to sample volume 4 Choose whether you want a Target Concentration cells mL or a Target Number of Cells and enter the...

Page 45: ...d 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 Cell Type Sample Images Bead Sample Images CD4 cells stained with Trypan Blue Jurkat Cells 5 µm Beads 10 µm Beads 15 µm Beads ...

Page 46: ...Chapter 8 Counting and Analysis Workflow 8001009 Cellometer Auto T4 User Manual Rev C 40 ...

Page 47: ... Cell Library EDITING CELL TYPES Cell types can be modified to meet your requirements using parameters available on the Cell Type Screen Cell Type Screen 1 Cell Type Name Identifies name given to the cell type 2 Detailed Description Contains cell type description 3 Save as New Cell Type Saves changes to parameter settings as new cell type 4 Lock from Future Editing Locks parameter settings so they...

Page 48: ...ackground 16 Do Not Decluster Clumps Indicates whether decluster enhancement settings are to be used 17 Use Template for Printing Indicates whether to use a print template 18 Set Default Allows users to select a default print template 19 Browse Allows users to browse for an existing print template 20 Edit Allows users to edit an existing print template 21 New Allows users to create a new print tem...

Page 49: ...d cell diameter For example the figure below shows counted cells for the same HT 29 image using different cell size parameter settings HT 29 cell count results were analyzed using different cell diameter parameter settings Figure a Cell Diameter Minimum size is too large Cells smaller than 15 µm are not counted as indicated by missing outlines Figure b Cell Diameter Maximum is too small Cells larg...

Page 50: ...0 60 available range is 0 04 stained cells have medium contrast to background to 0 90 stained cells have very low contrast to background Note It is a best practice to use the Very Dim Dead Cells check box in conjunction with other Trypan Blue Viability Parameters such as Enhancement Sensitivity or Uniformity values to avoid overcounting Dead cells with light blue centers To pick up dead cells with...

Page 51: ...luster Threshold Factor parameters may be used to separate clumpy cells i e connected cells for which it is difficult to distinguish clearly defined edges Default values for these parameters are 0 5 and 1 0 respectively For the majority of cell types the default values for these parameters are adequate to separate clumpy cells However some cell types such as MCF7 and PC12 contain cells having dim ...

Page 52: ...ng in MCF7 concentration of 8 3 x 105 cell mL DISABLING DECLUSTER FOR SMALL CELLS Examples of small size cells are mouse splenocytes lymphocytes thymocytes and PBMCs Their cell diameters are typically less than 7 µm The Auto T4 s focus position for really small cells may influence cell counting when the Decluster feature is enabled i e the Do not Decluster Clumps check box is not selected This fea...

Page 53: ... declustering is turned off by selecting the Do not Decluster Clumps check box more cells are counted Mouse thymocyte count results a with Decluster activated and b without Decluster However some samples contain clumpy cells as shown in figure a below Other samples have a very high cell concentration as shown in figure b For these samples the Decluster feature is activated i e Do not Decluster Clu...

Page 54: ...enable Counted check box to display overlay of green red and yellow outlines 10 Adjust cell type parameter settings and recount cells in the sample as necessary See Editing Cell Types on page 41 for details Creating Cell Type Manually 1 Select a cell type similar to the cells being used or choose the Initial Cell Type option from the drop down 2 Open the Cell Type menu from the menu bar and select...

Page 55: ...U812 L cell L1210 L929 large size cells LCL LLC PK1 Lncap Lymphoma M19 Macrophages MAGIC Malme 3M Mast MC32A MC32A B7 MC 38 MC 38 B7 MCF 10A MCF 10A4 MCF7 clumpy MCF7 non clumpy MCF7_clumpy MDA 231 MDA MB 231 MDA MB 435 MDA MB 468 MDBK medium size cells MEF Melan A Melan 3m Melanocyte Melanoma Mel STR Mel STV MES SA MG63 MM1S MMS MNT1 Mouse ES feeder cell MRC5 MSB1 MTA 9 MV411 N59 NCI 460 NCI H929...

Page 56: ...Chapter 9 Cell Types 8001009 Cellometer Auto T4 User Manual Rev C 50 ...

Page 57: ...ved to a text txt file Note Count results are saved by default to a single txt file in which rows logged with a date time stamp and Sample ID are appended each time data is saved Rows include key parameter settings used for the count 6 To view your data click the View Saved Data button or navigate to the folder and open the data file Alternatively you can use the New Data File and Open Data File o...

Page 58: ...e auto saving of data is set up results will be saved to the specified folder and file each time cells are counted Automatically Saving Images 1 Open the Options Menu and select the Save Options option 2 Click the Auto Save Sample Image check box to select it 3 Indicate whether you want to Save Raw Images saved as png files images can be loaded back into Auto T4 for reanalysis Save JPG Copies imag...

Page 59: ...age for Display option to load image for viewing or Load Image and Count option to load image and perform a count 2 Select the image file to load and click Open to return to the Home screen Note If the Intelligent File Load check box in the Counting Options screen available on the Options Menu is selected you will only need to select a file that ends in A as the remaining image will load automatic...

Page 60: ...avigate to a folder on the Operating Computer or network where data files are stored enter a file name and click Save Data is saved as an xlsx file and associated images if selected for export will be stored in a folder using the same name 2 To Save Counting Results to File click the Change File button to navigate to a folder on the Operating Computer or network where data files are stored enter a...

Page 61: ... e g Kimwipes 1 Dampen a fiber optic lint free wipe with IPA 2 Use the wipe to rub lightly on the outside of the instrument until it is visibly clean 3 Wait for the cleaning agent to evaporate before powering the instrument ON Should something break or be spilled inside the device power OFF the instrument and contact Nexcelom Support at 1 978 327 5340 or via email support nexcelom com ROUTINE MAIN...

Page 62: ...ess any functionality concerns at 1 978 327 5340 or via email support nexcelom com STORAGE When preparing the instrument for storage Always thoroughly clean the instrument cables and any of the accessories or consumables before storage Check for any damage and if possible re package the Instrument and Operating Computer Ensure that storage temperature and spatial requirements are met see Site Prep...

Page 63: ...wer Switch on instrument is set to the ON position Instrument displays software error messages The Auto T4 software has built in error messaging when functionality is operating improperly A few common error messages are listed below No instrument detected Background was saved but Dim XXX counts Error setting up Preview Could not read GPIO Values Running software in data analysis mode IF ERROR REQU...

Page 64: ...he instrument to request an upgrade The Power Cord or USB Connector Cable for the instrument has been lost Contact support nexcelom com with the serial number of the instrument to request replacements The cell type needed does not appear in the Cell Type drop down Cell types can be imported from either the Nexcelom Cell Library or from custom libraries stored in external network locations See Impo...

Page 65: ...om training and support or contact Nexcelom Support CONTACTING SUPPORT If there is a technical problem with your instrument or software contact Nexcelom Support Nexcelom Corporate Headquarters Nexcelom Bioscience 360 Merrimack Street Suite 200 Lawrence Massachusetts 01843 USA Phone 978 327 5340 Fax 978 327 5341 support nexcelom com San Diego Office Nexcelom Bioscience LLC 11100 Roselle Street Suit...

Page 66: ...Chapter 13 Nexcelom Support 8001009 Cellometer Auto T4 User Manual Rev C 60 ...

Page 67: ...al key advantages No potential clogging Ideal for fragile samples such as hepatocytes No washing No potential cross contamination Catalog Description Size Unit Image CHT4 PD100 002 Standard chamber thickness Packed in microscope slide boxes Individually QC Checked 4 year expiration Box of 100 slides for 200 counts 1 Box CHT4 PD100 003 Standard chamber thickness Packed in microscope slide boxes Ind...

Page 68: ...n Case of 900 slides Good for 1 800 counts 1 Case CHT4 SD100 314 Standard chamber thickness Packed with protective film 1 year expiration Case of 2 700 slides Good for 5 400 counts 3 Cases CHT4 SD100 514 Standard chamber thickness Packed with protective film 1 year expiration Case of 4 500 slides Good for 9 000 counts 5 Cases Visit the Nexcelom products cellometer disposable counting chambers page...

Page 69: ...5 µm polystyrene bead in Trypan Blue Solution Good for 200 counts 1 mL 4 Vials B10 02 020 10 µm polystyrene bead in Trypan Blue Solution Good for 200 counts 1 mL 4 Vials B15 02 010 15 µm Polystyrene bead in Trypan Blue Solution Good for 200 counts 1 mL 4 Vials Visit the Nexcelom products reagents page on our website for an up to date list and to purchase reagents for use with Cellometer instrument...

Page 70: ...Appendix A Consumables 8001009 Cellometer Auto T4 User Manual Rev C 64 ...

Page 71: ...A ITE emissions requirements Australia VCCI CISPR 32 2016 Class A ITE emissions requirements Japan EN 61000 3 2 2014 Limits for harmonic current emissions equipment input current up to and including 16A per phase EN 61000 3 3 2013 Limitation of voltage fluctuations and flicker in low voltage supply systemsfor equipment with rated current up to and including 16 A EN61326 1 2013 EMC requirements for...

Page 72: ...Appendix B Declaration of Conformity 8001009 Cellometer Auto T4 User Manual Rev C 66 ...

Page 73: ...rranty TERMS AND CONDITIONS The Nexcelom Bioscience LLC Terms and Conditions of Sale license agreement states the terms and conditions upon which Nexcelom Bioscience offers to license to you the software together with all related documentation The Software is licensed to you for use only in conjunction with Nexcelom s family of products In addition the original Cellometer software and any software...

Page 74: ...n 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 Buyer will not disclose the Nexcelom Proprietary Information to any third party or use it in any manner outside the scope of the license including 1 developing designin...

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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