A new method for yeast phagocytosis analysis by flow cytometry.

Area of Science:

• Immunology
• Microbiology
• Cell Biology

Background:

• Phagocytosis is a critical cellular process for immune defense and homeostasis.
• Accurate assessment of phagocytosis is essential for understanding host-pathogen interactions and immune cell function.
• Existing methods for quantifying phagocytosis may have limitations in distinguishing cell viability and yeast internalization.

Purpose of the Study:

• To develop and validate a novel flow cytometry-based method for assessing yeast phagocytosis.
• To enable accurate quantification of phagocytic activity, including cell viability and yeast internalization.
• To provide a robust tool for studying the complex interactions between phagocytes and fungal cells.

Main Methods:

• Development of a flow cytometry assay utilizing the quenching effect of propidium iodide on Sytox-Green fluorescence.
• Optimization of staining and gating strategies for accurate discrimination of cell populations.
• Validation of the method by comparing results with established techniques.

Main Results:

• The developed method accurately quantifies yeast phagocytosis by flow cytometry.
• It allows for the precise differentiation of live from dead phagocytes.
• The assay effectively distinguishes between internalized and non-internalized yeast cells while preserving yeast fluorescence within phagocytes, and characterizes different phagocyte-fungal cell associations.

Conclusions:

• This novel flow cytometry method provides a sensitive and accurate approach for assessing yeast phagocytosis.
• It offers significant advantages over existing techniques by simultaneously evaluating cell viability and yeast internalization.
• The method is valuable for research in immunology, microbiology, and cell biology, particularly for studying host-pathogen interactions.