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Three-Dimensional, Tomographic Super-Resolution Fluorescence Imaging of Serially Sectioned Thick Samples

Please always quote using this URN: urn:nbn:de:bvb:20-opus-134434
  • Three-dimensional fluorescence imaging of thick tissue samples with near-molecular resolution remains a fundamental challenge in the life sciences. To tackle this, we developed tomoSTORM, an approach combining single-molecule localization-based super-resolution microscopy with array tomography of structurally intact brain tissue. Consecutive sections organized in a ribbon were serially imaged with a lateral resolution of 28 nm and an axial resolution of 40 nm in tissue volumes of up to 50 \(\mu\)mx50\(\mu\)mx2.5\(\mu\)m. Using targetedThree-dimensional fluorescence imaging of thick tissue samples with near-molecular resolution remains a fundamental challenge in the life sciences. To tackle this, we developed tomoSTORM, an approach combining single-molecule localization-based super-resolution microscopy with array tomography of structurally intact brain tissue. Consecutive sections organized in a ribbon were serially imaged with a lateral resolution of 28 nm and an axial resolution of 40 nm in tissue volumes of up to 50 \(\mu\)mx50\(\mu\)mx2.5\(\mu\)m. Using targeted expression of membrane bound (m)GFP and immunohistochemistry at the calyx of Held, a model synapse for central glutamatergic neurotransmission, we delineated the course of the membrane and fine-structure of mitochondria. This method allows multiplexed super-resolution imaging in large tissue volumes with a resolution three orders of magnitude better than confocal microscopy.show moreshow less

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Metadaten
Author: Siddharth Nanguneri, Benjamin Flottmann, Heinz Horstmann, Mike Heilemann, Thomas Kuner
URN:urn:nbn:de:bvb:20-opus-134434
Document Type:Journal article
Faculties:Fakultät für Biologie / Theodor-Boveri-Institut für Biowissenschaften
Language:English
Parent Title (English):PLoS One
Year of Completion:2012
Volume:7
Issue:5
Pagenumber:e38098
Source:PLoS ONE 7(5): e38098. doi:10.1371/journal.pone.0038098
DOI:https://doi.org/10.1371/journal.pone.0038098
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Tag:architecture; in-vivo; localization; microscopy; proteins; rat calyx; reconstruction; resolution; transmission; ultrastructure
Release Date:2017/07/13
Licence (German):License LogoCC BY: Creative-Commons-Lizenz: Namensnennung