Photoswitching kinetics and phase-sensitive detection add discriminative dimensions for selective fluorescence imaging.

Content Provider	Semantic Scholar
Author	Querard, Jérôme Markus, Tal-Zvi Plamont, Marie-Aude Gauron, Carole Wang, Pengcheng Espagne, Agathe Volovitch, Michel Vriz, Sophie Croquette, Vincent Gautier, Arnaud Saux, Thomas Le Jullien, Ludovic
Copyright Year	2015
Abstract	Non-invasive separation-free protocols are attractive for analyzing complex mixtures. To increase selectivity, an analysis under kinetic control, through exploitation of the photochemical reactivity of labeling contrast agents, is described. The simple protocol is applied in optical fluorescence microscopy, where autofluorescence, light scattering, as well as spectral crowding presents limitations. Introduced herein is OPIOM (out-of-phase imaging after optical modulation), which exploits the rich kinetic signature of a photoswitching fluorescent probe to increase selectively and quantitatively its contrast. Filtering the specific contribution of the probe only requires phase-sensitive detection upon matching the photoswitching dynamics of the probe and the intensity and frequency of a modulated monochromatic light excitation. After in vitro validation, we applied OPIOM for selective imaging in mammalian cells and zebrafish, thus opening attractive perspectives for multiplexed observations in biological samples.
File Format	PDF HTM / HTML
DOI	10.1002/anie.201408985
PubMed reference number	25603793
Journal	Medline
Volume Number	54
Issue Number	9
Alternate Webpage(s)	http://www.ijb.univ-lorraine.fr/IMG/fichiers/fichiers/title_abstract_jullien.pdf
Alternate Webpage(s)	https://doi.org/10.1002/anie.201408985
Journal	Angewandte Chemie
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article