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

Scientific Reports
Jeroen VangindertaelHideaki Mizuno

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 fluo...Continue Reading

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Citations

Apr 24, 2016·Cellular and Molecular Life Sciences : CMLS·Anaël Chazeau, Grégory Giannone
Feb 10, 2018·Methods and Applications in Fluorescence·J VangindertaelK P F Janssen
Aug 10, 2016·Journal of Cell Science·Gautier FollainJacky G Goetz
May 2, 2020·ELife·Chih-Chieh Jay YuEdward S Boyden
Sep 8, 2018·ACS Nano·Herlinde De KeersmaeckerSusana Rocha

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Methods Mentioned

BETA
fluorescence microscopy
imaging techniques
fluorescence imaging
transgenic
confocal microscopy
imaging technique
super-resolution microscopy
electron
PCR

Software Mentioned

Adobe Photoshop
NASCA
Carl Zeiss Zen 2012
pcSOFI
ccPALM
dSTORM
Fluoview Viewer
Localizer
Olympus
CyGEL

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