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Context-dependent intravital imaging of therapeutic response using intramolecular FRET biosensors

Abstract

Intravital microscopy represents a more physiologically relevant method for assessing therapeutic response. However, the movement into an in vivo setting brings with it several additional considerations, the primary being the context in which drug activity is assessed. Microenvironmental factors, such as hypoxia, pH, fibrosis, immune infiltration and stromal interactions have all been shown to have pronounced effects on drug activity in a more complex setting, which is often lost in simpler two- or three-dimensional assays. Here we present a practical guide for the application of intravital microscopy, looking at the available fluorescent reporters and their respective expression systems and analysis considerations. Moving in vivo, we also discuss the microscopy set up and methods available for overlaying microenvironmental context to the experimental readouts. This enables a smooth transition into applying higher fidelity intravital imaging to improve the drug discovery process.

Type Journal
ISBN 1095-9130 (Electronic) 1046-2023 (Linking)
Authors Conway, J. R. W.; Warren, S. C.; Timpson, P.
Responsible Garvan Author A/Prof Paul Timpson
Publisher Name METHODS
Published Date 2017-09-01
Published Volume 128
Published Pages 78-94
Status Published in-print
DOI 10.1016/j.ymeth.2017.04.014
URL link to publisher's version https://www.ncbi.nlm.nih.gov/pubmed/28435000