Share Email Print
cover

Proceedings Paper

An automated wide-field time-gated optically sectioning fluorescence lifetime imaging multiwell plate reader for high-content analysis of protein-protein interactions
Author(s): Dominic Alibhai; Sunil Kumar; Douglas Kelly; Sean Warren; Yuriy Alexandrov; Ian Munro; James McGinty; Clifford Talbot; Edward J Murray; Frank Stuhmeier; Mark A. A. Neil; Chris Dunsby; Paul M. W. French
Format Member Price Non-Member Price
PDF $14.40 $18.00

Paper Abstract

We describe an optically-sectioned FLIM multiwell plate reader that combines Nipkow microscopy with wide-field time-gated FLIM, and its application to high content analysis of FRET. The system acquires sectioned FLIM images in <10 s/well, requiring only ~11 minutes to read a 96 well plate of live cells expressing fluorescent protein. It has been applied to study the formation of immature HIV virus like particles (VLPs) in live cells by monitoring Gag-Gag protein interactions using FLIM FRET of HIV-1 Gag transfected with CFP or YFP. VLP formation results in FRET between closely packed Gag proteins, as confirmed by our FLIM analysis that includes automatic image segmentation.

Paper Details

Date Published: 28 February 2011
PDF: 7 pages
Proc. SPIE 7904, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVIII, 79041Q (28 February 2011); doi: 10.1117/12.875135
Show Author Affiliations
Dominic Alibhai, Imperial College London (United Kingdom)
Sunil Kumar, Imperial College London (United Kingdom)
Douglas Kelly, Imperial College London (United Kingdom)
Sean Warren, Imperial College London (United Kingdom)
Yuriy Alexandrov, Imperial College London (United Kingdom)
Ian Munro, Imperial College London (United Kingdom)
James McGinty, Imperial College London (United Kingdom)
Clifford Talbot, Imperial College London (United Kingdom)
Edward J Murray, Pfizer Ltd. (United Kingdom)
Frank Stuhmeier, Pfizer Ltd. (United Kingdom)
Mark A. A. Neil, Imperial College London (United Kingdom)
Chris Dunsby, Imperial College London (United Kingdom)
Paul M. W. French, Imperial College London (United Kingdom)


Published in SPIE Proceedings Vol. 7904:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVIII
Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson; Thomas G. Brown, Editor(s)

© SPIE. Terms of Use
Back to Top