%0 Journal Article %1 pauli_targeted_2021 %A Pauli, Martin %A Paul, Mila M. %A Proppert, Sven %A Mrestani, Achmed %A Sharifi, Marzieh %A Repp, Felix %A Kürzinger, Lydia %A Kollmannsberger, Philip %A Sauer, Markus %A Heckmann, Manfred %A Sirén, Anna-Leena %D 2021 %J Communications Biology %K cctb computationalimageanalysis myown philipkollmannsberger %N 1 %P 1--13 %R 10.1038/s42003-021-01939-z %T Targeted volumetric single-molecule localization microscopy of defined presynaptic structures in brain sections %U https://www.nature.com/articles/s42003-021-01939-z %V 4 %X 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.

@article{pauli_targeted_2021, 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.}, added-at = {2021-04-14T16:10:03.000+0200}, author = {Pauli, Martin and Paul, Mila M. and Proppert, Sven and Mrestani, Achmed and Sharifi, Marzieh and Repp, Felix and Kürzinger, Lydia and Kollmannsberger, Philip and Sauer, Markus and Heckmann, Manfred and Sirén, Anna-Leena}, biburl = {https://www.bibsonomy.org/bibtex/2938a4b85401a0771a9ffaf13261d0ad0/philipk}, copyright = {2021 The Author(s)}, doi = {10.1038/s42003-021-01939-z}, file = {Full Text PDF:/Users/philip/Zotero/storage/RT2W7ZRR/Pauli et al. - 2021 - Targeted volumetric single-molecule localization m.pdf:application/pdf;Snapshot:/Users/philip/Zotero/storage/W6F5BCN5/s42003-021-01939-z.html:text/html}, interhash = {6c6417907d446ade4432103f5ea516ad}, intrahash = {938a4b85401a0771a9ffaf13261d0ad0}, issn = {2399-3642}, journal = {Communications Biology}, keywords = {cctb computationalimageanalysis myown philipkollmannsberger}, language = {en}, month = mar, note = {Number: 1Publisher: Nature Publishing Group}, number = 1, pages = {1--13}, timestamp = {2021-04-14T16:12:31.000+0200}, title = {Targeted volumetric single-molecule localization microscopy of defined presynaptic structures in brain sections}, url = {https://www.nature.com/articles/s42003-021-01939-z}, urldate = {2021-04-14}, volume = 4, year = 2021 }