1. Manuals
  2. Brands
  3. STEMCELL Manuals
  4. Measuring Instruments
  5. STEMvision
  6. Technical manual

STEMCELL STEMvision Technical Manual

Automated colony-forming unit (cfu) assay reader
Hide thumbs
Table of Contents

Advertisement

Quick Links

Download this manual
TECHNICAL MANUAL
STEMvision™
Automated Colony-Forming
Unit (CFU) Assay Reader

Advertisement

Table of Contents
loading
Need help?

Need help?

Do you have a question about the STEMvision and is the answer not in the manual?

Questions and answers

Subscribe to Our Youtube Channel

Summary of Contents for STEMCELL STEMvision

  • Page 1 TECHNICAL MANUAL STEMvision™ Automated Colony-Forming Unit (CFU) Assay Reader...

  • Page 3: Table Of Contents

    Editing Results ..........................33     STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.  TOLL-FREE PHONE...
  • Page 4 Appendix 1: Examples of “Lab” and “Parent” Human CFU Assay Report Forms ...... 40     STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. ...

  • Page 5: Safety Information And Warnings

    To reduce the risk of electrical shock, ensure that the instrument is unplugged prior to accessing openings other than the enclosure door. Do not allow fluids to enter the interior of the STEMvision™ instrument. In the event of such a spill, disconnect the power cable before cleaning. See section 8.4 for spill cleanup instructions.

  • Page 6: Introduction

    14-day CFU assays of bone marrow (BM), mobilized peripheral blood (MPB), or umbilical cord blood (CB) cells. STEMvision™ can also count the total number of colonies in a faster 7-day CFU assay of CB cells that has been developed for CB banks. A summary of human STEMvision™ Analysis Packages used for different human CFU assays is described in Table 1.
  • Page 7 Table 1: Summary of STEMvision™ Assays and MethoCult™ Media for Identifying Human CFU Types PRINTED COLONY STEMvision™ METHOCULT™ STEMvision™ ANALYSIS CFU ASSAY TISSUE ASSAY TYPES IDENTIFIED BY ASSAY TYPE MEDIUM PACKAGE REPORT DURATION STEMvision™ FIELD** FORM STEMvision™ Total CFUs Cord Blood MethoCult™...
  • Page 8 Table 2: Summary of STEMvision™ Assays and MethoCult™ Media for Identifying Mouse CFU Types COLONY COLONY CFU ASSAY METHOCULT™ STEMvision™ ANALYSIS STEMvision™ TISSUE TYPES SUPPORTED TYPES IDENTIFIED DURATION MEDIUM PACKAGE ASSAY TYPE FIELD* BY THE MEDIUM BY STEMvision™ BFU-E STEMvision™ Mouse Total MethoCult™...

  • Page 9: Stemvision™ System

    STEMCELL Technologies. Refer to Table 1 and Table 2 for a summary of the different types of CFU assays and recommended MethoCult™ media that are compatible with STEMvision™. For use with other MethoCult™...

  • Page 10: Unpacking And Setup

    4. Allow the inner chamber to warm up for about 10 minutes before inserting a SmartDish™ plate. Cultures should not be removed from the incubator until they are ready to be put inside STEMvision™ for imaging. Note: STEMvision™ is not specified for use as an incubator.
  • Page 11 Figure 1. Front View of the STEMvision™ Instrument STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. ...
  • Page 12 Figure 2. Back View of the STEMvision™ Instrument STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. ...
  • Page 13 Figure 3. Front View of the STEMvision™ Instrument with the Enclosure Door Open STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.

  • Page 14: Stemvision™ Acquisition Application

    4.0 STEMvision™ Acquisition Application The STEMvision™ Acquisition application is used to set up and control all aspects of CFU assay image acquisition. The application can only be run when the STEMvision™ instrument is turned on. Note: Do not turn off the STEMvision™ instrument while the STEMvision™ Acquisition application is running.

  • Page 15: Loading A Smartdish™ Plate

    1. Click the “Load Plate” button in the Control Panel to move the stage into the correct position for plate loading. 2. Open the STEMvision™ enclosure door (Figure 1F) and load a SmartDish™ plate onto the sample tray (Figure 3A). It is very important to ensure that the plate is properly inserted and sits completely flat on the tray.

  • Page 16: User, Sample, And Image Identification

    This is generated automatically and corresponds to the wells (A1 through B3) as seen in the Well Navigator dialog box in Figure 5. STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.
  • Page 17 Figure 5. Well Navigator Dialog Box Sample ID: The Sample ID information may be imported into the STEMvision™ Acquisition application via a CSV file or entered manually in the “Sample ID information” dialog box. Important: For human assays, if printed “Lab” or “Parent” CFU assay report forms will be required, the sample ID information must be entered via a CSV file.
  • Page 18 Sample information must be entered and saved in a CSV file before starting image acquisition. An example of a CSV file with 18 comma-separated columns is located on the desktop of the STEMvision™ computer upon installation of the device software.
  • Page 19 5. Click the “Done” button when all samples to be imaged have been added to the list. Note: Any Sample ID information added manually will be cleared on shutdown of the STEMvision™ Acquisition application. In order to save Sample ID Information, populate a CSV file as described under “Automatic Entry of Sample IDs from a Pre-Populated CSV File”.

  • Page 20: Image Identification

    Refer to section 4.4.2 for more information on Plate ID, Well ID, and Sample ID. STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.

  • Page 21: Setting Focus And Illumination

    The arrow keys on the PC’s keyboard or the scroll wheel on the mouse can be used for fine exposure adjustments. Figure 7. Setting the Illumination in the “Set Focus & Illumination” Dialog Box of the STEMvision™ Acquisition Application STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.
  • Page 22 If necessary, repeat the focus steps with a colony that better represents the average depth of colonies in the MethoCult™ medium. Figure 8. Setting the Focus in the “Set Focus & Illumination” Dialog Box of the STEMvision™ Acquisition Application STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485.

  • Page 23: Image Acquisition

    Image Acquisition The STEMvision™ Acquisition application produces a composite image of each culture well that comprises 35 separate camera images. The Human 7-Day CFU Assay and all mouse assays will result in a single bright-field image and the Human 14-Day CFU Assay will result in both a bright-field and a dark-field image which can be viewed separately or as a composite color image.

  • Page 24: Acquiring Images

    Acquisition application before starting image acquisition. Follow the steps outlined in section 5.0 to complete the image analysis. 4. After images of all cultures have been acquired, shut down STEMvision™ as described in section 3.3. STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.

  • Page 25: Stemvision™ Analyzer Application

    5.0 STEMvision™ Analyzer Application Note: The Programmed STEMvision™ Dongle must be inserted into one of the computer’s USB ports to run the STEMvision™ Analyzer application. The STEMvision™ Analyzer application is designed to analyze STEMvision™-acquired images offline and can be run independently of the STEMvision™ instrument. Different STEMvision™ Analysis Packages have been optimized to identify, classify, and count hematopoietic colonies from specific species and tissue types.

  • Page 26: User Interface Overview

    Start the STEMvision™ Analyzer application on the computer. For human assays, double-click the human icon and for mouse assays double-click the mouse icon on the desktop. The user interface of the human STEMvision™ Analyzer application is shown in Figure 10, and differs only slightly from the mouse STEMvision™ Analyzer application.

  • Page 27: Analysis Setup

    In order to generate “Lab” and “Parent” human CFU assay report forms, ensure that a CSV file is populated and imported into the STEMvision™ Acquisition application before acquiring images (section 4.4.2) and that a Sample ID has been defined for each well at the time of acquisition (section 4.6.1).

  • Page 28: Preview Of Result Images (Human And Mouse)

    (i.e. Plate ID folder). Select the images to be analyzed and then drag and drop them directly into the “Input Image” field of the STEMvision™ Analyzer application. If the desired analysis is one of the Color Human 14-Day algorithms, ensure that both the bright field and the dark field files are selected before dragging and dropping into the Input Image field.

  • Page 29: Analyzing Stemvision™ Acquired Images

    (7 or 14 days), and assay condition (with or without EPO) from the dropdown menu. To determine which STEMvision™ Analysis Package to use for your application or Assay Type, refer to Table 1 for human assays and Table 2 for mouse assays.

  • Page 30: Hl7 Data Export (Human Only)

    For further details on how to view results, see section 6.0. 5.3.3 HL7 Data Export (Human Only) STEMvision™ Analyzer for human CFU assays has the ability to export colony counts data to a Laboratory Information Management System (LIMS) in accordance with Health Level Seven International Standards (HL7).

  • Page 31: Viewing Results

    All results files for a given Plate ID can be accessed from the info.html file in the image folder. Viewing Human CFU Assay Report Forms The results of 7-Day and 14-Day CFU assays generated by STEMvision™ can be printed out in two different report formats: ...

  • Page 32: Viewing Individual Images

    The STEMvision™ Colony Marker application can be used independently of the STEMvision™ instrument and does not require the use of the STEMvision™ dongle. If desired, multiple images can be viewed at the same time by having several instances of this application open.
  • Page 33 ID number so that it can be identified in the image. For human assays it shows the summary of CFU type (i.e. CFU-E, BFU-E, CFU-G/M/GM or CFU-GEMM) and for mouse assays the summary of CFU size class (i.e. size class 1, 2, etc.) that has been scored by STEMvision™. ...
  • Page 34 Figure 14. Image and Analysis of a Human CFU Assay Using the STEMvision™ Colony Marker Application. This image is from a 14-day CFU assay on human BM cells that was analyzed with the STEMvision™ Color Human Bone Marrow 14-Day CFU Analysis Package. Red circles identify BFU-E-derived colonies; yellow circles identify CFU-G/M/GM-derived colonies;...
  • Page 35 For mouse assays, the colored circles around each colony correspond to the CFU size class that has been scored by STEMvision™. Four size classes (class 1 [smallest] to class 4 [largest]) are based on colony size and density (outlined below). This feature allows researchers to define size cutoffs so that very small colonies can be excluded from total colony counts if desired.

  • Page 36: Manually Scoring An Acquired Image

    “select” mode, press the “Home” key once again or select “Pan OFF” from the context menu. Manually Scoring an Acquired Image Images acquired using STEMvision™ can be scored manually using the STEMvision™ Colony Marker application. 1. Open the application and load the desired image file as described in section 6.2.2. There is no need to open the results file as well.

  • Page 37: Editing Results

    The CFU assay scoring results produced by the STEMvision™ Analysis application can be changed for individual colonies if desired using the STEMvision™ Colony Marker application. Note: In order to incorporate manual edits to STEMvision™ analysis results in the “Lab” or “Parent” CFU assay report forms, follow the steps outlined in section 6.5.

  • Page 38: Updating Sample Reports With Manually Modified Results

    Updating Sample Reports with Manually Modified Results The results of the STEMvision™ analysis can be manually reviewed and edited if desired using the Colony Marker application (see section 6.4). In order to incorporate modified STEMvision™ results in printed sample reports or HL7 transmissions, the modified results must first be imported by the Analyzer application.

  • Page 39: Creating A Summary Table Of Colony Counts In Microsoft® Excel

    7.0 Creating a Summary Table of Colony Counts in Microsoft® Excel The results of STEMvision™ analysis of multiple cultures can be tabulated in a Microsoft® Excel spreadsheet without viewing the images individually. Summary tables, as shown in Figure 18 and Figure 19, will be generated.

  • Page 40: Creating A Summary Table For Individual Plates Or Wells

    All results files that match the search criteria will be displayed in the dialog box. For example, to list all the results for images analyzed with the STEMvision™ Color Human Cord Blood 14-Day CFU Analysis Package, you may enter “*hCBDay14*” into the search field.

  • Page 41: Care And Maintenance

    Turn off the STEMvision™ instrument when it will not be used overnight or for a significant period of time during the day (section 3.3). As STEMvision™ is an enclosed system, it is sufficient to wipe the outside of the instrument occasionally with an appropriate non-corrosive decontaminant solution to keep it clean. If dilute bleach is used, wipe clean with water afterwards.

  • Page 42: Cleaning An External Spill

    Malfunction In the event of a malfunction, turn off STEMvision™ using the switch located on the back of the instrument (Figure 2G). Contact STEMCELL Technologies at techsupport@stemcell.com with a description of the failure so the appropriate corrective action can be determined.

  • Page 43: Specifications

    CAN/CSA C22.2 No. 61010-1-12 and UL standard No. 61010-1:2012 (safety requirements for electrical equipment for measurement, control, and laboratory use). If STEMvision™ is retired from regular use, return it to the shipping crate and store in conditions as specified. The STEMvision™ unit has the following symbol affixed to the back panel: This symbol denotes that the device should not be disposed of in municipal waste collection centers.

  • Page 44: Appendix 1: Examples Of "Lab" And "Parent" Human Cfu Assay Report Forms

    Figure 21. “Lab” Format of the Human CFU Assay Report Form for CB Cells STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.
  • Page 45 Microsoft Corporation. All other trademarks and registered trademarks are the property of their respective holders. While STEMCELL has made all reasonable efforts to ensure that the information provided by STEMCELL and its suppliers is correct, it makes no warranties or representations as to the accuracy or completeness of such information.
  • Page 48 TECHNICAL MANUAL STEMvision™ Automated Colony-Forming Unit (CFU) Assay Reader TOLL-FREE PHONE 1 800 667 0322 PHONE +1 604 877 0713 INFO@STEMCELL.COM TECHSUPPORT@STEMCELL.COM FOR GLOBAL CONTACT DETAILS VISIT WWW.STEMCELL.COM DOCUMENT #10000005577 VERSION 01 OCT 2019 | FOR INTERNAL USE: MATERIAL #29973...

Table of Contents