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Description

A movie from the time series stack described below (CIL:812) shows an amoeba (Dictyostelium discoideum) trying very hard to ingest a yeast cell (Saccharomyces cerevisiae) that is slightly too large. (The yeast cell is a mutant with an elongated bud. See note 1.) The amoeba is expressing two fluorescent fusion proteins: GFP-PLCdelta2 (note 2), which binds to phosphatidylinositol 4,5-bisphophate and labels the plasma membrane, and mRFP-LimEdelta (note 3), which labels actin filaments. Actin filaments shape the phagocytic cup at its leading edge and ring it in dynamic bands as it extends deep in the cell. The yeast is too long, and eventually the amoeba gives up, as indicated by the disappearance of the actin filaments. The yeast is promptly expelled with remarkable speed and force, and the excess membrane is converted into ruffles. The motile amoeba contacts the yeast anew, spreads along its length until reaching the end, then tries again to engulf it, but with no better luck. The similarities between a cell spreading on a substratum and forming a phagocytic cup are evident. Methods: Living cells were imaged using a Zeiss LSM 510 laser scanning confocal microscope with standard components and a Zeiss Plan-Apochromat 63x 1.4 N.A. DIC objective. Simultaneous fluorescence images (confocal) and DIC images were recorded from a single focal plane. Scans were performed every 3.89 seconds using a scan zoom of 3.5. References to reagents: 1: S. cerevisiae DDY-1306 (Kozminski, K.G., Chen, A.J., Rodal, A.A., Drubin, D.G. Mol. Biol. Cell 11:339-54, 2000). 2: Green Fluorescent Protein fused to the pleckstrin homology domain of phospholipase C delta-1 (Sauffer, T.P., Ahn, S., and Meyer, T., Curr. Biol. 8:343-346, 1998). 3: Monomeric Red Fluorescent Protein fused to a fragment of Dictyostelium LimE (Fischer, M., Haase, I., Simmeth, E., Gerisch, G., and Muller-Taubenberger, A., FEBS Lett. 577:227-232, 2004). The video received an Honorable Mention in the 2009 Olympus Bioscapes International Digital Imaging Competition and also may be viewed here: http://www.olympusbioscapes.com/gallery/2009/.

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
ingestion
egestion
digestion
response to food

Attribution

Name
Margaret Clarke
Publisheds
Clarke M, Engel U, Giorgione J, Müller-Taubenberger A, Prassler J, Veltman D, Gerisch G. Curvature recognition and force generation in phagocytosis. BMC Biol. 2010 Dec 29;8:154
Gerisch G, Ecke M, Schroth-Diez B, Gerwig S, Engel U, Maddera L, Clarke M. Self-organizing actin waves as planar phagocytic cup structures. Cell Adh Migr. 2009 Oct-Dec;3(4):373-82
Link
http://www.olympusbioscapes.com/galler...
Pubmed
21190565
19855162

Grouping

This image is part of a group.

Imaging

Image Type
fluorescence microscopy
Imaging Mode
recorded image
single-spot confocal microscopy
differential interference contrast microscopy
Parameters Imaged
single-spot confocal microscopy
Source of Contrast
fluorescence microscopy
distribution of a specific protein
boundaries between regions with different refractive index
Visualization Methods
fluorescent protein tag
mRFP-LimEΔ - probe for actin filaments
EGFP-PH-PLC-δ1 - binds phosphatidylinositol(4,5)bisphosphate
Processing History
recorded image
NIH Image J
Data Qualifiers
processed data

Sample Preparation

Methods
living tissue
Relation To Intact Cell
living tissue

Dimensions

Spatial Axis Image Size Pixel Size
X 512px ——
Y 512px ——
Time -- sec 0.17
*CIL – Cell Image Library accession number. Please use this to reference an image.