| Literature DB >> 33767432 |
Martin Pauli1,2, Mila M Paul1, Sven Proppert1,2, Achmed Mrestani1, Marzieh Sharifi1,2, Felix Repp1,2,3, Lydia Kürzinger1,2, Philip Kollmannsberger3, Markus Sauer4, Manfred Heckmann5, Anna-Leena Sirén6.
Abstract
Revealing the molecular organization of anatomically precisely defined brain regions is necessary for refined understanding of synaptic plasticity. Although three-dimensional (3D) single-molecule localization microscopy can provide the required resolution, imaging more than a few micrometers deep into tissue remains challenging. To quantify presynaptic active zones (AZ) of entire, large, conditional detonator hippocampal mossy fiber (MF) boutons with diameters as large as 10 µm, we developed a method for targeted volumetric direct stochastic optical reconstruction microscopy (dSTORM). An optimized protocol for fast repeated axial scanning and efficient sequential labeling of the AZ scaffold Bassoon and membrane bound GFP with Alexa Fluor 647 enabled 3D-dSTORM imaging of 25 µm thick mouse brain sections and assignment of AZs to specific neuronal substructures. Quantitative data analysis revealed large differences in Bassoon cluster size and density for distinct hippocampal regions with largest clusters in MF boutons.Entities:
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Year: 2021 PMID： 33767432 PMCID： PMC7994795 DOI： 10.1038/s42003-021-01939-z
Source DB: PubMed Journal: Commun Biol ISSN： 2399-3642