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.…
Author: | Siddharth Nanguneri, Benjamin Flottmann, Heinz Horstmann, Mike Heilemann, Thomas Kuner |
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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): | CC BY: Creative-Commons-Lizenz: Namensnennung |