Three-dimensional, tomographic super-resolution fluorescence imaging of serially sectioned thick samples
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 st...
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| Main Authors: | , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
May 25, 2012
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| In: |
PLOS ONE
Year: 2012, Volume: 7, Issue: 5 |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0038098 |
| Online Access: | Resolving-System, kostenfrei, Volltext: http://dx.doi.org/10.1371/journal.pone.0038098 Verlag, kostenfrei, Volltext: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0038098 
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| Author Notes: | Siddharth Nanguneri, Benjamin Flottmann, Heinz Horstmann, Mike Heilemann, Thomas Kuner |
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| 520 | |a 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 µm×50 µm×2.5 µm. 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. | ||
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| 650 | 4 | |a Mitochondria | |
| 650 | 4 | |a Tomography | |
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