Streamlined histone-based fluorescence lifetime imaging microscopy (FLIM) for studying chromatin organisation.

ABSTRACT: Changes in chromatin structure are key determinants of genomic responses. Thus, methods that enable such measurements are instrumental for investigating genome regulation and function. Here, we report further developments and validation of a streamlined method of histone-based fluorescence lifetime imaging microscopy (FLIM) that robustly detects chromatin compaction states in fixed and live cells, in 2D and 3D. We present a quality-controlled and detailed method that is simpler and faster than previous methods, and uses FLIMfit open-source software. We demonstrate the versatility of this chromatin FLIM through its combination with immunofluorescence and implementation in immortalised and primary cells. We applied this method to investigate the regulation of chromatin organisation after gen

SUBMITTER: Sherrard A 

PROVIDER: S-EPMC5898265 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature