Alternate header for print version


Licensing
Public Domain: This image is in the public domain and thus free of any copyright restrictions. However, as is the norm in scientific publishing and as a matter of courtesy, any user should credit the content provider for any public or private use of this image whenever possible. Learn more
tweet  
*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:812*  Cite 
Description

This time series stack 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/. A movie version of this stack is available in this library at CIL:12654.

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
Names
Margaret Clarke
Published
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
Published
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
Pubmed
21190565
Pubmed
19855162
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL812
Archival Resource Key (ARK)
ark:/b7295/w9cil812
Grouping This image is part of a group.
Sample Preparation
Methods
living tissue
Relation To Intact Cell
living tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 482px ——
Y 466px ——