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Quantitative evaluation of opsonin-independent phagocytosis by alveolar macrophages in monolayer using polystyrene
Abstract:
Macrophages can bind and engulf a variety of particles in the absence of specific opsonins. Polystyrene-type microspheres are often employed to quantitate opsonin-independent phagocytic activities of macrophages in vitro. Reliable measurement of this cell function, however, requires the ability of the investigator to distinguish between particles that are merely attached to the cell surface and those that are actually internalized. We have developed a simple, rapid, and reproducible method for quantitating phagocytosis using polystyrene microspheres and adherent alveolar macrophages. Basically, particles associated with macrophages after a given incubation time are microscopically quantitated on a cell-by-cell basis before and after toluene dissolution of external particles. Particle/macrophage values obtained after toluene treatment exclusively index phagocytosis.
Insights
Quantifying macrophage phagocytosis of microspheres is crucial. A new method using toluene distinguishes internalized particles from surface-bound ones, providing a reliable measure of opsonin-independent phagocytic activity.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Macrophages exhibit opsonin-independent phagocytosis of various particles.
- Polystyrene microspheres are commonly used to assess this macrophage function in vitro.
- Distinguishing internalized from surface-adherent particles is essential for accurate measurement.
Purpose of the Study:
- To develop a simple, rapid, and reproducible method for quantitating phagocytosis.
- To accurately measure opsonin-independent phagocytic activity of macrophages using polystyrene microspheres.
Main Methods:
- Utilized adherent alveolar macrophages and polystyrene microspheres.
- Quantitated particle association with macrophages microscopically on a cell-by-cell basis.
- Employed toluene dissolution to remove surface-adherent particles, isolating internalized ones.
Main Results:
- The toluene treatment effectively removed non-internalized particles.
- Post-treatment particle/macrophage values exclusively reflect phagocytosis.
- The method provides a reliable index of opsonin-independent phagocytosis.
Conclusions:
- The developed method accurately quantitates macrophage phagocytosis.
- This technique enhances the reliable measurement of opsonin-independent phagocytic activity.
- The toluene dissolution approach offers a straightforward way to assess particle internalization.