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Super-resolution mapping of glutamate receptors in C. elegans by confocal correlated PALM.

Abstract：

:Photoactivated localization microscopy (PALM) is a super-resolution imaging technique based on the detection and subsequent localization of single fluorescent molecules. PALM is therefore a powerful tool in resolving structures and putative interactions of biomolecules at the ultimate analytical detection limit. However, its limited imaging depth restricts PALM mostly to in vitro applications. Considering the additional need for anatomical context when imaging a multicellular organism, these limitations render the use of PALM in whole animals difficult. Here we integrated PALM with confocal microscopy for correlated imaging of the C. elegans nervous system, a technique we termed confocal correlated PALM (ccPALM). The neurons, lying below several tissue layers, could be visualized up to 10 μm deep inside the animal. By ccPALM, we visualized ionotropic glutamate receptor distributions in C. elegans with an accuracy of 20 nm, revealing super-resolution structure of receptor clusters that we mapped onto annotated neurons in the animal. Pivotal to our results was the TIRF-independent detection of single molecules, achieved by genetic regulation of labeled receptor expression and localization to effectively reduce the background fluorescence. By correlating PALM with confocal microscopy, this platform enables dissecting biological structures with single molecule resolution in the physiologically relevant context of whole animals.

journal_name

Sci Rep

journal_title

Scientific reports

authors

Vangindertael J,Beets I,Rocha S,Dedecker P,Schoofs L,Vanhoorelbeke K,Hofkens J,Mizuno H

doi

10.1038/srep13532

subject

Has Abstract

pub_date

2015-09-01 00:00:00

pages

13532

issn

2045-2322

pii

srep13532

journal_volume

5

pub_type

杂志文章

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