Thursday, 15 September 2016

Keeping score on centromeres with DRAQ5

Scientists in the lab of Dr. Ruth Wilkins at Health Canada have further developed their imaging flow cytometry methodologies for radiation biodosimetry, to detect the presence of aberrant dicentric chromosomes as a further tool to improve the ability to triage exposed individuals in the event of a large-scale radiological or nuclear emergency, such as the Fukushima disaster. 

This work builds on their earlier publications using the far-red cell permeant DNA probe DRAQ5 (BioStatus Limited) to detect micronuclei as a facile signature for radiation damage in peripheral blood leukocytes on the Amnis Imagestream imaging flow cytometer (EMD-Millipore).

Briefly, in this most recent work, they isolate leukocytes from fresh blood, stimulate the cells with PHA and interfere with proper chromosome separation with colcemid to promote formation of dicentric chromosomes. The chromosomes are released into suspension and labeled with a centromere-specific fluorescent PNA probe and DRAQ5 for DNA content. The labeled chromosomes are analysed by imaging flow cytometry to measure the centromere count per chromosome.

Reference: Beaton-Green LA, Rodrigues MA, Lachapelle S, Wilkins RC. Foundations of identifying individual chromosomes by imaging flow cytometry with applications in radiation biodosimetry. Methods. 2016 Aug 12.

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