TY  - JOUR
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  - Kuner, Thomas
A1  - Klevanski, Maja
A1  - Heilemann, Mike
T1  - Visualizing synaptic multi-protein patterns of neuronal tissue with DNA-assisted single-molecule localization microscopy
T2  - Frontiers in synaptic neuroscience
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 (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.
KW  - single-molecule localization microscopy
KW  - super-resolution microscopy
KW  - DNA-PAINT
KW  - neuronal synapse
KW  - multiplexing
KW  - Exchange PAINT
KW  - semi-thin brain tissue sections
KW  - tissue imaging
Y1  - 2021
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/61761
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-617614
SN  - 1663-3563
VL  - 13
IS  - art. 671288
SP  - 1
EP  - 9
PB  - Frontiers Research Foundation
CY  - Lausanne
ER  -