SyncRGB-FLIM: synchronous fluorescence imaging of red, green and blue dyes enabled by ultra-broadband few-cycle laser excitation and fluorescence lifetime detection
File(s)boe-10-4-1891.pdf (5.95 MB)
Published version
Author(s)
Type
Journal Article
Abstract
We demonstrate for the first time that an ultra-broadband 7 femtosecond (fs) few-cycle laser can be used for multicolor nonlinear imaging in a single channel detection geometry, when employing a time-resolved fluorescence detection scheme. On a multi-chromophore-labelled cell sample we show that the few-cycle laser can efficiently excite the multiple chromophores over a >400 nm two-photon absorption range. By combining the few-cycle laser excitation with time-correlated single-photon counting (TCSPC) detection to record two-photon fluorescence lifetime imaging microscopy (FLIM) images, the localization of different chromophores in the cell can be identified based on their fluorescence decay properties. The novel SyncRGB-FLIM multi-color bioimaging technique opens the possibility of real-time protein-protein interaction studies, where its single-scan operation translates into reduced laser exposure of the sample, resulting in more photoprotective conditions for biological specimens.
Date Issued
2019-04-01
Date Acceptance
2019-02-06
Citation
Biomedical Optics Express, 2019, 10 (4), pp.1891-1891
ISSN
2156-7085
Publisher
Optica Publishing Group
Start Page
1891
End Page
1891
Journal / Book Title
Biomedical Optics Express
Volume
10
Issue
4
Copyright Statement
© 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement
License URL
Identifier
http://dx.doi.org/10.1364/boe.10.001891
Publication Status
Published
Date Publish Online
2019-03-18