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

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).

• 2021

• Clarke, Margaret
• Maddera, Lucinda

Preparation: whole mounted tissue
Relation to intact cell: whole mounted tissue
Item type: fluorescence microscopy; fluoresence quenching
Imaging mode: recorded image; single-spot confocal microscopy
Parameter imaged: single-spot confocal microscopy
Source of contrast: fluorescence microscopy
Visualization methods: Fluorescein (FITC); PA-mRFP; PA-GFP
Processing history: unprocessed raw data
Data qualification: Raw;spatialmeasurements

• Cell Image Library Group ID: 3691

• Dictyostelium discoideum

• Amoeboid cell
• Cell by organism
• Eukaryotic cell
• Eukaryotic Protist
• Food vacuole
• Phagocytic vesicle

• Dictyostelium discoideum strain AX2–214
• Digestion
• Response to food

• data
• image

• No linguistic content; Not applicable

Samplenumber: 7326

Doi: https://doi.org/10.6075/J04J0D3H

Source data

• Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL7326

Reference

• Clarke M, Maddera L, Harris RL, Silverman PM. F-pili dynamics by live-cell imaging. Proc Natl Acad Sci U S A. 2008 Nov 18;105(46):17978-81. https://doi.org/10.1073/pnas.0806786105
• PubMed article: https://www.ncbi.nlm.nih.gov/pubmed/?term=19004777

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2022-08-12