Immunophenotyping and DNA content analysis of acetone-fixed cells

Area of Science:

• Cell Biology
• Hematology
• Oncology
• Cytometry

Background:

• Acetone is a fixative used in static cytometry but rarely in flow cytometry due to cell shrinkage.
• Traditional flow cytometry methods may not fully capture physical cell characteristics alongside phenotypic and DNA content data.

Purpose of the Study:

• To introduce and validate the flow acetone staining technique (FAST) for comprehensive cellular analysis.
• To demonstrate FAST's utility in identifying diverse proliferating cell subpopulations within heterogeneous samples.

Main Methods:

• Utilizing acetone fixation at approximately 8°C to preserve cellular integrity and membrane permeability.
• Concurrent analysis of light-scatter properties, surface/nuclear immunophenotypes, and DNA content using flow cytometry.

Main Results:

• FAST effectively preserves cellular physical features and immunophenotype.
• The technique allows for accurate stoichiometric DNA staining and content measurement.
• Diverse subpopulations of proliferating cells were identified by immunophenotype and cell cycle status.

Conclusions:

• FAST offers a valuable approach for concurrent analysis of physical, phenotypic, and DNA content in flow cytometry.
• The technique is suitable for cell biology and hematology/oncology studies, enabling detailed subpopulation analysis and DNA ploidy assessment.