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Related Concept Videos

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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Flow Cytometric Characterization of Murine B Cell Development
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Immune Cell Phenotyping Using Flow Cytometry.

A Graham Pockley1,2, Gemma A Foulds1, Julie A Oughton3

  • 1John van Geest Cancer Research Center, Nottingham Trent University, Nottingham, United Kingdom.

Current Protocols in Toxicology
|November 3, 2015
PubMed
Summary
This summary is machine-generated.

Fluorescent immunophenotyping, including flow cytometry, identifies cell types and changes using fluorescent antibodies. This method rapidly analyzes millions of cells, offering detailed insights into cell populations and phenotypes.

Keywords:
cell analysisflow cytometryimmune phenotypingmethodologytoxicology

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Area of Science:

  • Cell Biology
  • Immunology
  • Biotechnology

Background:

  • Fluorescent immunophenotyping identifies distinct cell subpopulations in heterogeneous samples.
  • Flow cytometry offers rapid, objective analysis of millions of cells, characterizing physical and fluorescent properties.
  • This technique is crucial for understanding cell populations in tissues and single-cell suspensions.

Purpose of the Study:

  • To describe basic procedures for direct and indirect immunofluorescent staining of lymphoid cells.
  • To provide essential information on antibody selection, fluorochrome choice, and data analysis standardization.
  • To highlight new technologies for in-depth cell subpopulation and phenotype interrogation.

Main Methods:

  • Direct and indirect immunofluorescent staining of surface and intracellular proteins.
  • Protocols for dead cell resolution and cell fixation.
  • Guidance on antibody titration, fluorochrome selection, spectral compensation, and controls.

Main Results:

  • Standardized protocols enable accurate identification and quantification of cell subpopulations.
  • Detailed information on optimizing antibody-fluorochrome combinations and data acquisition.
  • New technologies offer enhanced depth in analyzing cell phenotypes.

Conclusions:

  • Fluorescent immunophenotyping, particularly flow cytometry, is a powerful tool for cell analysis.
  • Standardized methods and careful optimization are key to reliable results.
  • Emerging technologies promise even greater resolution in cellular analysis.