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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Related Experiment Video

Updated: Jun 22, 2026

Analysis of Cell Suspensions Isolated from Solid Tissues by Spectral Flow Cytometry
11:08

Analysis of Cell Suspensions Isolated from Solid Tissues by Spectral Flow Cytometry

Published on: May 5, 2017

EMS mutant spectra generated by multi-parameter flow cytometry.

Stephen B Keysar1, Michael H Fox

  • 1Cell and Molecular Biology Graduate Program, Colorado State University, Fort Collins, CO 80523-1618, United States.

Mutation Research
|May 26, 2009
PubMed
Summary
This summary is machine-generated.

This study developed a flow cytometry mutation assay (FCMA) to detect mutations in cell surface proteins like CD59. The assay identified ethyl methanesulfonate (EMS) induced mutations, revealing distinct mutant spectra compared to radiation.

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Area of Science:

  • Cell Biology
  • Genetics
  • Toxicology

Background:

  • CHO A(L) cells possess human chromosome 11 with GPI-linked (CD59, CD90) and non-GPI-linked (CD98, CD44, CD151) proteins.
  • Flow Cytometry Mutation Assay (FCMA) detects CD59 gene mutations via antibody fluorescence absence.
  • Simultaneous mutation analysis of multiple cell surface proteins generates comprehensive mutant spectra.

Purpose of the Study:

  • To generate a mutant spectrum for ionizing radiation using simultaneous mutation detection.
  • To analyze mutations induced by ethyl methanesulfonate (EMS) in CHO A(L) cells.
  • To refine the FCMA for improved detection of CD59 mutants.

Main Methods:

  • Utilized CHO A(L) cell line with human chromosome 11.
  • Applied FCMA to measure mutations in CD59, CD44, CD90, CD98, and CD151.
  • Induced mutations using ethyl methanesulfonate (EMS) and ionizing radiation.
  • Sorted single cells and generated clonal populations for stable phenotype analysis.
  • Used fluorescently labeled aerolysin (FLAER) to identify GPI-deficient mutants.

Main Results:

  • EMS treatment resulted in intermediate CD59 staining, with sorted clones showing stable CD59 expression.
  • Nearly all isolated clones exhibited mutations in CD59 only.
  • Approximately 60% of CD59-negative clones were GPI mutants, likely due to pigA gene mutations.
  • EMS-induced small mutations differed significantly from radiation-induced large deletions.
  • FCMA mutant regions were adjusted from 1% to 10% to encompass most CD59 mutants.

Conclusions:

  • FCMA is effective for generating mutant spectra of various cell surface proteins.
  • EMS induces distinct mutation patterns (small mutations) compared to ionizing radiation (large deletions).
  • GPI biosynthesis pathway mutations (e.g., pigA) contribute to CD59-negative phenotypes.
  • Adjusting FCMA parameters enhances sensitivity for detecting CD59 mutations.