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Quantification of apoptosis and necrosis by flow cytometry
M G Ormerod1, X M Sun, D Brown
1MRC Toxicology Unit, Carshalton, Surrey, England.
Acta Oncologica (Stockholm, Sweden)
|January 1, 1993
Summary
This study introduces a novel flow cytometry method to distinguish and quantify viable normal, apoptotic, and necrotic cells. The technique uses Hoechst 33342 and propidium iodide dyes for cell death analysis.
Area of Science:
- Cell Biology
- Biochemistry
- Immunology
Background:
- Apoptosis and necrosis are critical cell death pathways.
- Quantifying these processes is essential for biological research.
- Existing flow cytometry methods for apoptosis detection have limitations.
Purpose of the Study:
- To develop and validate a novel flow cytometry method for distinguishing and quantifying viable normal, apoptotic, and necrotic cells.
- To apply this method to different cell types and experimental conditions.
Main Methods:
- Incubation of cells with Hoechst 33342 and propidium iodide dyes.
- Flow cytometry analysis utilizing UV excitation.
- Characterization of three distinct cell populations based on fluorescence emission.
Main Results:
- Viable normal cells exhibited low blue and low red fluorescence.
- Apoptotic cells fluoresced bright blue.
- Necrotic (dead) cells fluoresced red due to propidium iodide uptake.
Conclusions:
- The developed method effectively quantifies and separates viable normal, apoptotic, and necrotic cells.
- This technique provides a valuable tool for studying cell death mechanisms.
- The method is applicable to various cell types and experimental models.