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Flow Cytometry01:23

Flow Cytometry

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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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CellTrace™ Violet Flow Cytometric Assay to Assess Cell Proliferation.

Marina B Lemieszek1, Scott D Findlay2,3, Gabrielle M Siegers4

  • 1Department of Obstetrics and Gynecology, University of Alberta, Edmonton, AB, Canada.

Methods in Molecular Biology (Clifton, N.J.)
|June 23, 2022
PubMed
Summary

CellTrace™ Violet (CTV) is a reliable method for tracking cell proliferation and monitoring multiple cell divisions. This study details its application across various cell types and protocol optimization insights.

Keywords:
Cell divisionCell trackingDilution of dyeDivision trackingFlow cytometryProliferation assayProliferation modeling

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Area of Science:

  • Cell Biology
  • Immunology
  • Biotechnology

Background:

  • Cell proliferation is a fundamental biological process.
  • Accurate tracking of cell division is crucial for understanding cellular dynamics.
  • Existing methods may have limitations in sensitivity or multi-generational tracking.

Purpose of the Study:

  • To describe the application of CellTrace™ Violet (CTV) for cell proliferation tracking.
  • To demonstrate CTV's utility in diverse cell types.
  • To share key observations from protocol optimization for CTV staining.

Main Methods:

  • CellTrace™ Violet staining of cells.
  • Monitoring fluorescence intensity reduction over cell divisions.
  • Quantification of proliferation using flow cytometry.
  • Application in various cell types and co-culture systems.

Main Results:

  • CellTrace™ Violet permanently labels cellular proteins, enabling fluorescence tracking.
  • Each cell division halves the fluorescence intensity per daughter cell.
  • CTV allows for multi-generational proliferation monitoring.
  • Successful application demonstrated across different cell types.

Conclusions:

  • CellTrace™ Violet is a versatile and effective tool for quantifying cell proliferation.
  • Protocol optimization is essential for reliable results in diverse cellular contexts.
  • CTV facilitates the study of cell division dynamics and population tracking.