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

CIL:7653*  Cite 
Description

Phagocytosis of bacteria by Dictyostelium discoideum amoebae. These Dictyostelium cells are expressing GFP-2FYVE, which binds to phosphatidylinositol 3-phosphate [PI(3)P], and mRFP-LimEdelta, which binds to actin filaments. The bacteria are E. coli that had been incubated with an inhibitor of cytokinesis, producing elongated forms. Phosphoinositides help to confer specific identities to compartments along the endocytic pathway. PI(3)P identifies early endosomes. It associates with phagosomes ~1 minute after uptake and persists as acidification occurs and digestion begins. Note that the long bacteria do not bind the green PI(3)P marker until after they have been fully internalized. The changing morphology of the long bacteria within the labeled phagosomes reveals the action of digestive enzymes. Three successive time series of 100 frames were collected in a single focal plane using a Zeiss LSM510 confocal microscope with a 63x DIC objective lens. The frames are separated by 3.94 seconds. There was a space of 6 seconds between the first two time series and a space of 9 seconds between the second and third time series. The scaling (pixel size) was 0.10 x 0.10 microns. The images were exported as TIFs using Zeiss LSM 5 Image Examiner and saved both as a TIF stack and as an AVI movie (available at CIL:7654) using NIH ImageJ. The movie spans 20 minutes; it plays at 7.5 fps, 30 times actual speed. 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
response to food
ingestion
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/W9CIL7653
Archival Resource Key (ARK)
ark:/b7295/w9cil7653
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
PA-GFP
Processing History
unprocessed raw data
Data Qualifiers
raw, unprocessed data
suitable for spatial measurements
Sample Preparation
Methods
whole mounted tissue
Relation To Intact Cell
whole mounted tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 519px 0.1µm
Y 512px 0.1µm