Monitoring internalization of Histoplasma capsulatum by mammalian cell lines using a fluorometric microplate assay
AJ Scott, JP Woods - Medical mycology, 2000 - academic.oup.com
AJ Scott, JP Woods
Medical mycology, 2000•academic.oup.comWe have developed a fluorometric microtiter plate assay to quantify the internalization of
Histoplasma capsulatum yeasts by macrophages. The assay utilizes the fluorescent dye
Calcofluor White to label the yeast cell wall and the vital dye trypan blue, which does not
enter viable macrophages, to quench fluorescence of extracellular labeled yeasts. Murine
RAW 264.· 7 cells showed more efficient internalization of strain G217B yeasts than human
U937 cells. Both cell lines exhibited a dependence upon actin, and, to a lesser degree …
Histoplasma capsulatum yeasts by macrophages. The assay utilizes the fluorescent dye
Calcofluor White to label the yeast cell wall and the vital dye trypan blue, which does not
enter viable macrophages, to quench fluorescence of extracellular labeled yeasts. Murine
RAW 264.· 7 cells showed more efficient internalization of strain G217B yeasts than human
U937 cells. Both cell lines exhibited a dependence upon actin, and, to a lesser degree …
Abstract
We have developed a fluorometric microtiter plate assay to quantify the internalization of Histoplasma capsulatum yeasts by macrophages. The assay utilizes the fluorescent dye Calcofluor White to label the yeast cell wall and the vital dye trypan blue, which does not enter viable macrophages, to quench fluorescence of extracellular labeled yeasts. Murine RAW 264.·7 cells showed more efficient internalization of strain G217B yeasts than human U937 cells. Both cell lines exhibited a dependence upon actin, and, to a lesser degree, microtubules, in G217B uptake.
Oxford University Press