PT  - JOURNAL ARTICLE
AU  - Pauli, Martin
AU  - Paul, Mila M.
AU  - Proppert, Sven
AU  - Sharifi, Marzieh
AU  - Repp, Felix
AU  - Kollmannsberger, Philip
AU  - Sauer, Markus
AU  - Heckmann, Manfred
AU  - Sirén, Anna-Leena
TI  - Targeted volumetric single-molecule localization microscopy of defined presynaptic structures in brain sections
AID  - 10.1101/568279
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 568279
4099  - http://biorxiv.org/content/early/2019/03/05/568279.short
4100  - http://biorxiv.org/content/early/2019/03/05/568279.full
AB  - Revealing the molecular organization of anatomically precisely defined brain regions is necessary for the refined understanding of synaptic plasticity. Although, three-dimensional (3D) single-molecule localization microscopy can provide the required molecular resolution, single-molecule 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 aberration-free 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 enables 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.