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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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Related Experiment Video

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Modified Annexin V/Propidium Iodide Apoptosis Assay For Accurate Assessment of Cell Death
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Modified Annexin V/Propidium Iodide Apoptosis Assay For Accurate Assessment of Cell Death

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Protocol for Apoptosis Assay by Flow Cytometry Using Annexin V Staining Method.

Imayavaramban Lakshmanan1, Surinder K Batra1

  • 1Biochemistry and Molecular Biology, Nebraska Medical Center, Omaha, USA.

Bio-Protocol
|July 19, 2016
PubMed
Summary

This assay quantifies apoptosis by detecting phosphatidylserine exposure on cell membranes using Annexin V staining. Propidium iodide is used to distinguish between apoptotic and necrotic cells via flow cytometry.

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Area of Science:

  • Cell Biology
  • Biochemistry

Background:

  • Apoptosis is a critical biological process involving programmed cell death.
  • Distinguishing apoptotic cells from necrotic cells is essential for accurate biological analysis.

Purpose of the Study:

  • To describe an assay for quantifying the number of cells undergoing apoptosis.
  • To detail the methodology for differentiating apoptotic and necrotic cells.

Main Methods:

  • Cells are stained with Annexin V and propidium iodide.
  • Flow cytometry is utilized for quantitative analysis.
  • The assay relies on detecting phosphatidylserine exposure on the outer cell membrane.

Main Results:

  • The assay successfully identifies and quantifies cells in apoptosis.
  • Annexin V binding indicates phosphatidylserine translocation.
  • Propidium iodide staining differentiates necrotic cells with compromised membranes.

Conclusions:

  • This Annexin V and propidium iodide staining assay provides a reliable method for apoptosis detection.
  • Flow cytometry analysis enables accurate differentiation of apoptotic and necrotic cell populations.