STEMvision™ - Automated Counter for CFU Assays

Automated and Standardized Counting of CFU Assays of Human and Mouse Hematopoietic Progenitor Cells

STEMvision™ is a bench-top instrument and computer system designed specifically for automated imaging and counting of hematopoietic colonies in the colony-forming unit (CFU) assay . Instead of manually identifying and counting colonies using a microscope, the user simply loads a SmartDish™ culture plate containing up to 6 individual CFU assays in MethoCult™ medium into the automated CFU counter. The instrument captures an image of each 35 mm well, which is then analyzed using specialized software for each type of assay. By using an automated CFU counter to standardize colony counting, cord blood banks, transplant labs and research labs can ensure that their CFU assay results are accurate and reproducible.

How to Standardize Hematopoietic Colony Counting with STEMvision™

Watch step-by-step guidelines on how to use STEMvision™ for automated imaging and counting of hematopoietic colonies in CFU assays.

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Why Use STEMvision™?

  • Achieve more accurate colony counts than with manual counting.
  • Software reduces inter- and intra-individual and laboratory variation in colony counting.
  • Standardized software reduces the time it takes to train new students and staff to recognize and count colonies.
  • Culture wells are digitally imaged for permanent record keeping.

STEMvision™ for Automated CFU Counting of Human and Mouse CFU Assays

Human 7 Day

Cell Types:

Cord Blood

MethoCult™ Media:

Image Output:

Black and white images of entire 35 mm culture well and marked colonies

Data Output:

Total combined counts of all colonies

Imaging Time:

30 seconds

Analysis Time:

30 seconds

Human 14 Day

Cell Types:

  • Cord Blood
  • Bone Marrow
  • Mobilized Peripheral Blood

Image Output:

Color images highlighting hemoglobinized erythroid colonies with subclasses of colonies marked

Data Output:

Differential counts of colony subtypes (CFU-E, BFU-E, CFU-G/M/GM, CFU-GEMM)

Imaging Time:

1 minute

Analysis Time:

1 minute

Mouse 7-14 Day

Cell Types:

Bone Marrow

Image Output:

Black and white images of entire 35 mm culture well with 4 size classes of colonies marked

Data Output:

Total combined counts of all colonies in lineage-specific MethoCult™ media

Imaging Time:

30 seconds

Analysis Time:

30 seconds

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Data

Automated CFU counter, STEMvision™, images and analyses human CFU colonies

Figure 1. Representative STEMvision™ Images Showing Colonies Derived from Human Cord Blood Progenitors After 7 Days of Culture in MethoCult™ Express, and from Human Cord Blood, Bone Marrow and Mobilized Peripheral Blood Progenitors After 14 Days of Culture in MethoCult™ Optimum

These images have been analyzed by STEMvision™ Human (A) 7-Day and (B-D) 14-Day Analysis Packages. Green circles identify individual colonies in a 7-day cord blood CFU assay that scores total CFUs only (A). Orange and red circles identify erythroid colonies, yellow circles identify myeloid colonies and blue circles identify mixed colonies in 14-day cord blood (B), bone marrow (C) and mobilized peripheral blood (D) CFU assays. Erythroid and mixed colonies that contain hemoglobinized cells are shown in true red color.



Automated CFU counter, STEMvision™, images and analyses mouse CFU colonies

Figure 2. Representative STEMvision™ Images Showing Colonies Derived from Mouse Bone Marrow Progenitors After 12 Days of Culture in MethoCult™ GF M3434, MethoCult™ GF M3534 or MethoCult™ SF M3436 Media

Images of mouse bone marrow cells cultured in (A) MethoCult™ GF M3434, (B) MethoCult™ GF M3534 and (C) MethoCult™ SF M3436 were acquired using STEMvision™. The corresponding STEMvision™ Mouse Analysis Package (i.e. Total CFU Analysis Package (A), Myeloid CFU Analysis Package (B) or Erythroid CFU Analysis Package (C)) was used to analyze each image. Red circles identify the smallest colonies - size class 1, yellow circles - size class 2, blue circles - size class 3 and orange circles identify the largest colonies - size class 4.

Applications and Regulatory Compliance

In addition to the quantification of progenitor cell numbers and subtypes in normal hematopoietic samples, the CFU assay may be used to compare the relative progenitor cell function of modified samples. Through this use, the CFU assay and STEMvision™ have become important tools for the development of new cell and gene therapies, including the assay of CD34+ cells that have been expanded, modified or edited (e.g. with CRISPR-Cas9) ex vivo.

Higher compliance environments such as quality control or manufacturing departments may face additional requirements such as compliance to FDA Title 21 CFR Part 11 regulations on electronic records. The STEMvision™ software suite is designed for research use only (RUO) applications. However, STEMvision™ can support compliance to 21 CFR Part 11 regulations through the addition of the STEMvision™ 21 CFR Part 11 Compliance Software Add-On (Catalog #500-0110).

The STEMvision™ 21 CFR Part 11 Compliance Software Add-On provides numerous features to increase the security of electronic records, including:

  • User authentication and restricted access levels
  • Electronic signatures (human analysis packages only)
  • Logging of software events and the creation of audit trails

For more details, please refer to the STEMvision™ product page, STEMvision™ Manual, or contact us. Learn more about our approach and commitment to quality and how we can support your regulatory and compliance by visiting our Quality at STEMCELL page.


CFU Assay Resources

Explore our resources for information and support on setting up, optimizing, and standardizing the CFU assay.

Wallchart
Identification of Colonies Derived from Human Hematopoietic Progenitors
Video
How to Set Up Hematopoietic Colony-Forming Unit (CFU) Assays
Brochure
STEMvision™ Automated and Standardized Counting of Mouse Bone Marrow CFU Assays
Brochure
STEMvision™ Automated and Standardized Counting of CFU Assays of Human Hematopoietic Cells
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