TY  - INPR
A1  - Narayanasamy, Kaarjel K.
A1  - Stojic, Aleksandar
A1  - Li, Yunqing
A1  - Saß, Steffen
A1  - Hesse, Marina R.
A1  - Deußner-Helfmann, Nina Sabine
A1  - Dietz, Marina
A1  - Klevanski, Maja
A1  - Kuner, Thomas
A1  - Heilemann, Mike
T1  - Visualizing multi-protein patterns at the synapse of neuronal tissue with DNA-assisted single-molecule localization microscopy
T2  - bioRxiv
N2  - 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. Using antibodies labeled with short DNA oligonucleotides, multiple targets are visualized successively by sequential exchange of fluorophore-labeled complementary oligonucleotides present in the imaging buffer. The structural integrity of the tissue is maintained owing to only a single labelling step during sample preparation. Multiple targets are imaged using a single laser wavelength, minimizing chromatic aberration. This method proved robust for multi-target imaging in semi-thin tissue sections, paving the way towards structural cell biology with single-molecule super-resolution microscopy.
Y1  - 2021
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/72864
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-728645
IS  - 2021.02.23.432306
ER  -