Fluorescence in Situ Hybridization and Nuclear Translocation Events Quantified by Imaging Flow Cytometry.
Year: 2019
Type: article
Abstract: Imaging flow cytometry with its ability to perform image analysis on up to 10 spectrally separated but spatially correlated images of the same cell in statistically robust cell population sizes has steadily seen an increase in the number of applications and the use of this platform in various research settings to a point where it is now commonly found in flow and/or image cytometry shared resource centers. One of the originally anticipated applications of imaging flow cytometry was in the area of fluorescent in situ hybridization (FISH), a technique which, due to its manual evaluation of slide-based sample preparations, suffers from a relatively low sensitivity of detection. A second early application was in the area of quantifying nuclear translocation events of transcription factors such as NFkB and NFAT which conventionally had been analyzed by microscopy or molecular approaches such as western blot or gel shift assays of nuclear protein extractions. Practical examples and considerations of using imaging flow cytometry to assess FISH and nuclear translocation of NFkB and NFAT will be presented with a discussion of the challenges of sample preparation and data analysis faced with each of these applications and the experience at Roswell Park with incorporating this analysis platform in a shared resource setting. Supported by NIH 1S10OD018048, R50CA211108, and NCI Cancer Center Support Grant to Roswell Park P30-CA016056.
Author: Hans Minderman
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W3004918644