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PLoS One. 2012;7(5):e38098. doi: 10.1371/journal.pone.0038098. Epub 2012 May 25.

Three-dimensional, tomographic super-resolution fluorescence imaging of serially sectioned thick samples.

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  • 1Institute of Anatomy and Cell Biology, Heidelberg University, Heidelberg, Germany.

Abstract

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.

PMID:
22662272
[PubMed - indexed for MEDLINE]
PMCID:
PMC3360663
Free PMC Article
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