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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:7328*  Cite 
Description

A movie of phagosome 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, a phagosome is neutralized prior to exocytosis of the indigestible yeast carcass; that process is visualized here. The amoebae are also expressing two other fluorescent markers that are barely 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 (available at CIL:7327)and saved as a stack in ImageJ; they are also shown as a movie (.avi) at 10fps. For related experiments and further details, see Clarke et al., PLoS ONE 5:e8585 (2010).

Biological Sources
NCBI Organism Classification
Dictyostelium discoideum
Cell Type
cell by organism
eukaryotic cell
Eukaryotic Protist
amoeboid cell
Cell Line
Dictyostelium discoideum strain AX2–214
Cellular Component
phagocytic vesicle
food vacuole
Biological Context
Biological Process
digestion
cellular response to food
Attribution
Names
Margaret Clarke
Lucinda Maddera
Published
Clarke et al., PLoS ONE 5:e8585 (2010)
Pubmed
20052281
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL7328
Archival Resource Key (ARK)
ark:/b7295/w9cil7328
Grouping This image is part of a group.
Imaging
Image Type
fluorescence microscopy
fluoresence quenching
Image Mode
recorded image
single-spot confocal microscopy
Parameters Imaged
single-spot confocal microscopy
Source of Contrast
fluorescence microscopy
Visualization Methods
Fluorescein (FITC)
PA-mRFP
PA-GFP
Processing History
unprocessed raw data
Data Qualifiers
raw, unprocessed data
Sample Preparation
Methods
whole mounted tissue
Relation To Intact Cell
whole mounted tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 232px ——
Y 170px ——