Please use this identifier to cite or link to this item: http://ahro.austin.org.au/austinjspui/handle/1/19387
Title: Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy.
Authors: Parslow, Adam C;Clayton, Andrew H A;Lock, Peter;Scott, Andrew M
Affiliation: Department of Molecular Imaging and Therapy, Austin Health, Heidelberg, Victoria, Australia
Tumour Targeting Laboratory, Olivia Newton-John Cancer Research Institute, Heidelberg, Victoria, Australia
School of Cancer Medicine, La Trobe University, Melbourne, Victoria, Australia
Department of Medical Oncology, Olivia Newton John Cancer Wellness and Research Centre, Austin Health, Heidelberg, Victoria, Australia
Department of Medicine, Austin Health, The University of Melbourne, Heidelberg, Victoria, Australia
Centre for Micro-Photonics, Faculty of Science, Engineering and Technology, Swinburne University of Technology
LIMS Bioimaging Facility, La Trobe Institute for Molecular Science, La Trobe University
Issue Date: 2-Aug-2018
EDate: 2018-08-02
Citation: Journal of visualized experiments : JoVE 2018; 138
Abstract: Confocal microscopy provides an accessible methodology to capture sub-cellular interactions critical for the characterization and further development of pre-clinical agents labeled with fluorescent probes. With recent advancements in antibody based cytotoxic drug delivery systems, understanding the alterations induced by these agents within the realm of receptor aggregation and internalization is of critical importance. This protocol leverages the well-established methodology of fluorescent immunocytochemistry and the open source FIJI distribution of ImageJ, with its inbuilt autocorrelation and image mathematical functions, to perform spatial image correlation spectroscopy (ICS). This protocol quantitates the fluorescent intensity of labeled receptors as a function of the beam area of the confocal microscope. This provides a quantitative measure of the state of target molecule aggregation on the cell surface. This methodology is focused on the characterization of static cells with potential to expand into temporal investigations of receptor aggregation. This protocol presents an accessible methodology to provide quantification of clustering events occurring at the cell surface, utilizing well established techniques and non-specialized imaging apparatus.
URI: http://ahro.austin.org.au/austinjspui/handle/1/19387
DOI: 10.3791/57164
ORCID: 0000-0002-6656-295X
PubMed URL: 30124657
Type: Journal Article
Video-Audio Media
Research Support, Non-U.S. Gov't
Appears in Collections:Journal articles

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