Visualizing synaptic multi-protein patterns of neuronal tissue with DNA-assisted single-molecule localization microscopy

  • The development of super-resolution microscopy (SRM) has widened our understanding of biomolecular structure and function in biological materials. Imaging multiple targets within a single area would elucidate their spatial localization relative to the cell matrix and neighboring biomolecules, revealing multi-protein macromolecular structures and their functional co-dependencies. SRM methods are, however, limited to the number of suitable fluorophores that can be imaged during a single acquisition as well as the loss of antigens during antibody washing and restaining for organic dye multiplexing. We report the visualization of multiple protein targets within the pre- and postsynapse in 350–400 nm thick neuronal tissue sections using DNA-assisted single-molecule localization microscopy (SMLM). In a single labeling step, antibodies conjugated with short DNA oligonucleotides visualized multiple targets by sequential exchange of fluorophore-labeled complementary oligonucleotides present in the imaging buffer. This approach avoids potential effects on structural integrity when using multiple rounds of immunolabeling and eliminates chromatic aberration, because all targets are imaged using a single excitation laser wavelength. This method proved robust for multi-target imaging in semi-thin tissue sections with a lateral resolution better than 25 nm, paving the way toward structural cell biology with single-molecule SRM.

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Author:Kaarjel K. Narayanasamy, Aleksandar Stojic, Yunqing Li, Steffen Saß, Marina Hesse, Nina Sabine Deußner-Helfmann, Marina DietzORCiDGND, Thomas Kuner, Maja Klevanski, Mike HeilemannORCiDGND
URN:urn:nbn:de:hebis:30:3-617614
DOI:https://doi.org/10.3389/fnsyn.2021.671288
ISSN:1663-3563
Parent Title (English):Frontiers in synaptic neuroscience
Publisher:Frontiers Research Foundation
Place of publication:Lausanne
Document Type:Article
Language:English
Date of Publication (online):2021/06/17
Date of first Publication:2021/06/17
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2021/08/03
Tag:DNA-PAINT; Exchange PAINT; multiplexing; neuronal synapse; semi-thin brain tissue sections; single-molecule localization microscopy; super-resolution microscopy; tissue imaging
Volume:13
Issue:art. 671288
Page Number:9
First Page:1
Last Page:9
HeBIS-PPN:487443632
Institutes:Biochemie, Chemie und Pharmazie / Biochemie und Chemie
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Sammlungen:Universitätspublikationen
Open-Access-Publikationsfonds:Biochemie, Chemie und Pharmazie
Licence (German):License LogoCreative Commons - Namensnennung 4.0