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Description
A movie that shows phagosome acidification and neutralization visualized using FITC-yeast. Amoebae of Dictyostelium discoideum were fed living budding yeast (S. cerevisiae strain TH2-1B) that had been labeled with fluorescein isothiocyanate (FITC, 0.25 mg/ml). The amoebae in phosphate buffer (pH 6.4) were examined 1 ½ hours later using a Zeiss LSM510 laser scanning confocal microscope; frames were collected in a single focal plane at 5-second intervals. The yeast fluoresce brightly in the extracellular medium, but are dim in acidic phagosomes because FITC fluorescence is quenched at acidic pH. At the end of the endocytic pathway, phagosomes are neutralized and the indigestible yeast carcass is exocytosed. This time series shows fluctuations in fluorescence intensity just prior to exocytosis of a yeast. That yeast is promptly phagocytosed by another cell, and the yeast grows dim as the phagosome is acidified. Meanwhile a second yeast in the same cell grows brighter, presumably in preparation for exocytosis. The amoebae are also expressing two other fluorescent markers, which are faintly visible, mRFP-LimE-delta, which labels actin filaments, and VatM-GFP, a subunit of the V-ATPase. Three hundred frames from the time series were exported as .tif images and saved as a stack in ImageJ (and available at CIL:7625); they are shown here as a movie (.avi) at 10fps. For related experiments and further details, see Clarke et al., PLoS ONE 5:e8585 (2010). Research Data Curation Program, UC San Diego, La Jolla, 92093-0175 (https://lib.ucsd.edu/rdcp) Clarke, Margaret; Maddera, Lucinda (2021). CIL:7326, Dictyostelium discoideum, cell by organism, eukaryotic cell, amoeboid cell, Eukaryotic Protist. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J04J0D3H
Type
image
Identifier
ark:/20775/bb57067608
Language
No linguistic content
Subject
Dictyostelium discoideum strain AX2–214 Response to food Digestion Eukaryotic cell Phagocytic vesicle Amoeboid cell Cell by organism Food vacuole Eukaryotic Protist Dictyostelium discoideum Cell Image Library Group ID: 3691
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