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A rapid single cell sorting verification method using plate-based image cytometry.

Eric S Zigon1, Suzanne M Purseglove2, Vasilis Toxavidis1

  • 1Flow Cytometry Core, Beth Israel Deaconess Medical Center, Boston, Massachusetts.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|August 3, 2018
PubMed
Summary
This summary is machine-generated.

This study presents a rapid method for verifying single cell sorting using the Celigo Image Cytometer. The new technique quickly confirms single cells in wells, improving flow cytometry and cell-based assays.

Keywords:
Celigocell line developmentfluorescence activated cell sorting (FACS)image cytometrymonoclonalitysingle cell sorting

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Area of Science:

  • Cell Biology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Single cell sorting is crucial for monoclonality and homogenous cell populations.
  • Manual verification of single cells is time-consuming and labor-intensive.
  • A rapid and accurate method is needed to verify single cell sorting efficiency.

Purpose of the Study:

  • To develop and validate a rapid single cell sorting verification method.
  • To assess the utility of the Celigo Image Cytometer for this purpose.
  • To improve the optimization of fluorescence-activated cell sorting (FACS) instruments.

Main Methods:

  • Jurkat cells and fluorescent beads were stained with Calcein AM.
  • Cells and beads were sorted into 96-well microplates using MoFlo Astrios EQ.
  • Whole well images were acquired and analyzed using the Celigo Image Cytometer within 8 minutes.

Main Results:

  • The Celigo Image Cytometer rapidly acquired and analyzed images of sorted cells.
  • The method accurately verified the presence of single cells in microplate wells.
  • Analysis time was significantly reduced compared to manual microscopy.

Conclusions:

  • The developed method offers a fast and accurate approach for single cell sorting verification.
  • This technique can optimize FACS instrument performance in core facilities.
  • Improved verification enhances the reliability of downstream biological assays.