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Flow Cytometry

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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Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry

Measuring Caspase Activity Using a Fluorometric Assay or Flow Cytometry

Published on: March 24, 2023

Detecting Cleaved Caspase-3 in Apoptotic Cells by Flow Cytometry.

Lisa C Crowley¹, Nigel J Waterhouse^1,2,3

¹Apoptosis and Cytotoxicity Laboratory, Mater Research, Translational Research Institute, Woolloongabba, Brisbane, Queensland 4102, Australia.

Cold Spring Harbor Protocols

|November 3, 2016

Summary

This summary is machine-generated.

This study details a flow cytometry method to quantify apoptosis by detecting cleaved caspase-3. This approach reliably identifies cells undergoing or having undergone programmed cell death.

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Area of Science:

Cell Biology
Biochemistry

Background:

Apoptosis is a regulated cell death process orchestrated by caspases, a family of cysteine proteases.
Active caspases cleave specific substrates, generating detectable fragments during apoptosis.

Purpose of the Study:

To outline a protocol for quantifying apoptosis using flow cytometry.
To highlight the detection of cleaved caspase-3 as a reliable marker for apoptotic cells.

Main Methods:

Utilizing antibodies specific to caspase-cleaved fragments.
Employing flow cytometry for the quantification of cells exhibiting caspase cleavage.
Focusing on the detection of cleaved caspase-3, a key executioner caspase.

Main Results:

Antibodies recognizing cleaved caspase fragments enable specific labeling of apoptotic cells.
Flow cytometry allows for the quantification of these labeled cells.
Detection of cleaved caspase-3 serves as a robust indicator of apoptosis.

Conclusions:

Flow cytometry combined with cleaved caspase-3 detection offers a reliable method for quantifying apoptosis.
This technique facilitates the identification and measurement of cells undergoing programmed cell death.