Visualizing multi-protein patterns at the synapse of neuronal tissue with DNA-assisted single-molecule localization microscopy

摘要: The development of super-resolution microscopy (SRM) has widened our understanding biomolecular structure and function in biological materials. Imaging multiple targets within a single area would elucidate their spatial localization relative to the cell matrix neighboring biomolecules, revealing multi-protein macromolecular structures functional co-dependencies. SRM methods are, however, limited number suitable fluorophores that can be imaged during acquisition as well loss antigens antibody washing restaining for organic dye multiplexing. We report visualization protein pre- postsynapse 350-400 nm thick neuronal tissue sections using DNA-assisted single-molecule microscopy. Using antibodies labeled with short DNA oligonucleotides, are visualized successively by sequential exchange fluorophore-labeled complementary oligonucleotides present imaging buffer. structural integrity is maintained owing only labelling step sample preparation. Multiple laser wavelength, minimizing chromatic aberration. This method proved robust multi-target semi-thin sections, paving way towards biology