A new quantitative fluorimetric assay for phagocytosis of bacteria.
We describe a new quantitative fluorimetric assay for phagocytosis of bacteria. A suspension of fluorescein-labelled bacteria (Micrococcus lysodeikticus) is mixed and incubated with phagocytes. After fixation with paraformaldehyde, the excess non-phagocytosed and adsorbed bacteria are lysed with a solution of lysozyme in phosphate-buffered saline. After washing of the phagocytes, the fluorescence of those that have ingested the labelled bacteria is measured with a spectrofluorimeter. We report results obtained with different types of phagocytes which show that this method allows sensitivity, saturation and kinetic studies and the calculation of a phagocytic index.
Vray
B
B
Laboratory of Microbiology and Immunology, Faculty of Medicine, Free University of Brussels.
Hoebeke
J
J
Saint-Guillain
M
M
Leloup
R
R
Strosberg
A D
AD
D000818:Animals; D007158:Immunologic Techniques; D008837:Micrococcus / Q000276:immunology*; D009504:Neutrophils / Q000502:physiology*; D010586:Phagocytes / Q000502:physiology*; D010587:Phagocytosis; D011817:Rabbits; D013050:Spectrometry, Fluorescence / Q000379:methods*