High-throughput quantification of early stages of phagocytosis

Yeo, Jeremy Changyu, Wall, Adam Alexander, Stow, Jennifer Lea and Hamilton, Nicholas Ahti (2013) High-throughput quantification of early stages of phagocytosis. Biotechniques, 55 3: 115-124. doi:10.2144/000114075


Author Yeo, Jeremy Changyu
Wall, Adam Alexander
Stow, Jennifer Lea
Hamilton, Nicholas Ahti
Title High-throughput quantification of early stages of phagocytosis
Journal name Biotechniques   Check publisher's open access policy
ISSN 0736-6205
1940-9818
Publication date 2013-09-01
Year available 2013
Sub-type Article (original research)
DOI 10.2144/000114075
Open Access Status Not yet assessed
Volume 55
Issue 3
Start page 115
End page 124
Total pages 7
Place of publication New York, United States
Publisher Informa Healthcare
Language eng
Abstract Phagocytosis—the engulfment of cells and foreign bodies—is an important cellular process in innate immunity, development, and disease. Quantification of various stages of phagocytosis, especially in a rapid screening fashion, is an invaluable tool for elucidating protein function during this process. However, current methods for assessing phagocytosis are largely limited to flow cytometry and manual image-based assays, providing limited information. Here, we present an image-based, semi-automated phagocytosis assay to rapidly quantitate three distinct stages during the early engulfment of opsonized beads. Captured images are analyzed using the image-processing software ImageJ and quantified using a macro. Modifications to this method allowed quantification of phagocytosis only in fluorescently labeled transfected cells. Additionally, the time course of bead internalization could be measured using this approach. The assay could discriminate perturbations to stages of phagocytosis induced by known pharmacological inhibitors of filamentous actin and phosphoinositol-3-kinase. Our methodology offers the ability to automatically categorize large amounts of image data into the three early stages of phagocytosis within minutes, clearly demonstrating its potential value in investigating aberrant phagocytosis when manipulating proteins of interest in drug screens and disease.
Keyword Receptor-Mediated Phagocytosis
Macrophage Phagocytosis
Assay
Maturation
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2014 Collection
Institute for Molecular Bioscience - Publications
 
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