Development of a high-throughput γ-H2AX assay based on imaging flow cytometry.

ABSTRACT:

Background

Measurement of γ-H2AX foci levels in cells provides a sensitive and reliable method for quantitation of the radiation-induced DNA damage response. The objective of the present study was to develop a rapid, high-throughput γ-H2AX assay based on imaging flow cytometry (IFC) using the ImageStream®^X Mk II (ISX) platform to evaluate DNA double strand break (DSB) repair kinetics in human peripheral blood cells after exposure to ionizing irradiation.

Methods

The γ-H2AX protocol was developed and optimized for small volumes (100 μL) of human blood in Matrix™ 96-tube format. Blood cell lymphocytes were identified and captured by ISX INSPIRE™ software and analyzed by Data Exploration and Analysis Software.

Results

Dose- and time-dependent γ-H2AX levels cor

SUBMITTER: Lee Y 

PROVIDER: S-EPMC6704696 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature