Assessment of cell viability by flow cytometric analysis using DNase exclusion

Area of Science:

• Cell Biology
• Molecular Biology
• Biotechnology

Background:

• Assessing cell viability is crucial in biological research and drug development.
• Existing methods may have limitations in accurately distinguishing between viable and non-viable cells based on DNA content.

Purpose of the Study:

• To develop a novel method for the selective measurement of DNA distributions in viable cell populations.
• To establish a reliable index of cell viability using DNase treatment.

Main Methods:

• Developed a method utilizing DNase (deoxyribonuclease) to degrade DNA in non-viable cells, which lose membrane integrity.
• Employed 4'-6-diamidino-2-phenylindole (DAPI) staining and flow cytometry for DNA distribution analysis before and after DNase treatment.
• Quantified cell viability by assessing the percentage of cells retaining DNA after DNase exposure.

Main Results:

• The DNase exclusion test effectively differentiates between viable and non-viable cells based on DNA content.
• The method demonstrated applicability in analyzing spontaneous cell death, drug-induced cell death, and hyperthermia-induced cell death.
• Optimal conditions for DNase treatment and the DNase exclusion test were established.

Conclusions:

• The developed DNase-based method offers a selective and accurate approach for measuring DNA in viable cells.
• This technique serves as a valuable tool for assessing cell viability and analyzing cell death mechanisms.
• The DNase exclusion test is versatile and applicable to various cell death scenarios.