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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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Hauptverfasser: Nanguneri, Siddharth (VerfasserIn) , Flottmann, Benjamin (VerfasserIn) , Horstmann, Heinz (VerfasserIn) , Heilemann, Mike (VerfasserIn) , Kuner, Thomas (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: May 25, 2012
In: PLOS ONE
Year: 2012, Jahrgang: 7, Heft: 5
ISSN:1932-6203
DOI:10.1371/journal.pone.0038098
Online-Zugang: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
Volltext
Verfasserangaben:Siddharth Nanguneri, Benjamin Flottmann, Heinz Horstmann, Mike Heilemann, Thomas Kuner
Beschreibung
Zusammenfassung: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.
Beschreibung:Gesehen am 02.11.2018
Beschreibung:Online Resource
ISSN:1932-6203
DOI:10.1371/journal.pone.0038098

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