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A flow cytometric method for the measurement of phagocytosis by polymorphonuclear leucocytes
Abstract:
A new method for the measurement of phagocytosis of Candida albicans by human polymorphonuclear leucocytes (PMN) is described using a fluorescence activated cell sorter. We have used acridine orange to discriminate between PMN which have internalised yeast particles and those which have not. This method allows accurate measurement of particle phagocytosis as an event distinct from particle adherence. It also permits detailed examination of the kinetics of phagocytosis, the study of which is likely to be of value in the investigation of diseases where abnormalities of PMN function are suspected.
Insights
This study introduces a novel fluorescence-activated cell sorter method to accurately measure the phagocytosis of Candida albicans by human polymorphonuclear leucocytes (PMN). This technique distinguishes internalization from adherence, aiding in the study of PMN function in disease.
Area of Science:
- Immunology
- Cell Biology
- Microbiology
Background:
- Phagocytosis is a critical immune process involving the engulfment of pathogens by immune cells.
- Human polymorphonuclear leucocytes (PMN) are key players in the innate immune response, particularly against fungal infections like Candida albicans.
- Accurate measurement of phagocytosis is essential for understanding immune function and diagnosing related diseases.
Purpose of the Study:
- To develop and describe a novel, accurate method for quantifying Candida albicans phagocytosis by human PMN.
- To differentiate between particle internalization and adherence during phagocytosis.
- To enable detailed kinetic studies of phagocytosis for investigating immune cell dysfunction.
Main Methods:
- Utilized a fluorescence-activated cell sorter (FACS) for high-throughput analysis.
- Employed acridine orange staining to differentiate between PMN that have internalized yeast particles and those that have not.
- Established a method to measure phagocytosis as a distinct event from particle adherence.
Main Results:
- The described FACS method accurately quantifies the phagocytosis of Candida albicans by human PMN.
- The technique successfully distinguishes between yeast particle internalization and adherence to PMN.
- The method allows for detailed kinetic analysis of the phagocytic process.
Conclusions:
- A new, precise method for measuring PMN phagocytosis of Candida albicans has been established.
- This technique facilitates the study of phagocytosis kinetics and provides insights into PMN function.
- The method holds potential value for investigating diseases associated with abnormal PMN function.