Literature DB >> 30536013

Rapid Imaging of BCL-2 Family Interactions in Live Cells Using FLIM-FRET.

Elizabeth J Osterlund1,2, Nehad Hirmiz2,3, Christian Tardif4, David W Andrews5,6.   

Abstract

The Bcl-2 proteins control cell death via interchanging interactions within the Bcl-2 family. Fluorescence lifetime imaging microscopy (FLIM) is used to detect Förster resonance energy transfer (FRET) between two fluorescent-fusion proteins in live cells. FLIM-FRET has been used to detect specific interactions and their disruption, for Bcl-2 family proteins. To date, this has been possible only in low throughput, due to the time required for serial data acquisition. We developed an automated optical system with eight parallel detectors for rapid and efficient data collection. Here we describe how to use this system for FLIM-FRET imaging of Bcl-2 family protein interactions in a 384-well plate format.

Entities:  

Keywords:  BH3 mimetic; Bcl-2 family; FLIM Hyperspectral; FLIM-FRET; Fluorescence lifetime imaging microscopy; High throughput; mCerulean3

Mesh:

Substances:

Year:  2019        PMID: 30536013     DOI: 10.1007/978-1-4939-8861-7_19

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


  4 in total

1.  Using FLIM-FRET for Characterizing Spatial Interactions in the Spindle.

Authors:  Stephanie C Ems-McClung; Claire E Walczak
Journal:  Methods Mol Biol       Date:  2022

2.  Measuring Small-molecule Inhibition of Protein Interactions in Live Cells Using FLIM-FRET.

Authors:  James M Pemberton; Qian Liu; David W Andrews
Journal:  Bio Protoc       Date:  2019-10-20

3.  Fluorescence lifetime imaging microscopy: fundamentals and advances in instrumentation, analysis, and applications.

Authors:  Rupsa Datta; Tiffany M Heaster; Joe T Sharick; Amani A Gillette; Melissa C Skala
Journal:  J Biomed Opt       Date:  2020-05       Impact factor: 3.170

4.  Efficacy and specificity of inhibitors of BCL-2 family protein interactions assessed by affinity measurements in live cells.

Authors:  Elizabeth J Osterlund; Nehad Hirmiz; James M Pemberton; Adrien Nougarède; Qian Liu; Brian Leber; Qiyin Fang; David W Andrews
Journal:  Sci Adv       Date:  2022-04-20       Impact factor: 14.957

  4 in total

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