Journal
PLOS ONE
Volume 6, Issue 2, Pages -Publisher
PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pone.0015820
Keywords
-
Categories
Funding
- European Union [STRP 013880, MRTN-2005-019481]
- German Ministry of Education and Research, BMBF [FKZ 13N10077]
- German Research Foundation, DFG [SFB 854, FOR 521 HA 3498/1-1/2]
- Spanish Ministry of Science and Innovation MICINN/FEDER [BFU2008-03288]
- Consejeria de Sanidad, Junta de Comunidades de Castilla-La Mancha [JI4002R2]
- Centro Regional de Investigaciones Biomedicas
Ask authors/readers for more resources
Time-domain Fluorescence Lifetime Imaging Microscopy (FLIM) is a remarkable tool to monitor the dynamics of fluorophore-tagged protein domains inside living cells. We propose a Wide-Field Multi-Parameter FLIM method (WFMP-FLIM) aimed to monitor continuously living cells under minimum light intensity at a given illumination energy dose. A powerful data analysis technique applied to the WFMP-FLIM data sets allows to optimize the estimation accuracy of physical parameters at very low fluorescence signal levels approaching the lower bound theoretical limit. We demonstrate the efficiency of WFMP-FLIM by presenting two independent and relevant long-term experiments in cell biology: 1) FRET analysis of simultaneously recorded donor and acceptor fluorescence in living HeLa cells and 2) tracking of mitochondrial transport combined with fluorescence lifetime analysis in neuronal processes.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available