Fluorescence lifetime imaging nanoscopy for measuring Förster resonance energy transfer in cellular nanodomains.

TitleFluorescence lifetime imaging nanoscopy for measuring Förster resonance energy transfer in cellular nanodomains.
Publication TypeJournal Article
Year of Publication2019
AuthorsTardif C, Nadeau G, Labrecque S, Côté D, Lavoie-Cardinal F, De Koninck P
JournalNeurophotonics
Volume6
Issue1
Pagination015002
Date Published2019 Jan
ISSN2329-423X
Abstract

Microscopy methods used to measure Förster resonance energy transfer (FRET) between fluorescently labeled proteins can provide information on protein interactions in cells. However, these methods are diffraction-limited, thus do not enable the resolution of the nanodomains in which such interactions occur in cells. To overcome this limitation, we assess FRET with an imaging system combining fluorescence lifetime imaging microscopy with stimulated emission depletion, termed fluorescence lifetime imaging nanoscopy (FLIN). The resulting FRET-FLIN approach utilizes immunolabeling of proteins in fixed cultured neurons. We demonstrate the capacity to discriminate nanoclusters of synaptic proteins exhibiting variable degrees of interactions with labeled binding partners inside dendritic spines of hippocampal neurons. This method enables the investigation of FRET within nanodomains of cells, approaching the scale of molecular signaling.

DOI10.1117/1.NPh.6.1.015002
Alternate JournalNeurophotonics
PubMed ID30746389
PubMed Central IDPMC6354015

Funding

Our research endeavors are made possible by the following agencies:

Canadian Institutes of Health Research - Instituts de recherche en santé du Canada Fonds de recherche du Québec – Nature et technologies (FRQNT)Fonds de la recherche en santé du Québec NARSAD Natural Sciences and Engineering Research Council of Canada (NSERC) - Conseil de recherche en sciences naturelles et en génie du Canada (CRSNG)innovation.caHuman Frontier Science Program