Three-Dimensional Imaging of Biological Tissue by Cryo X-Ray Ptychography

Abstract High-throughput three-dimensional cryogenic imaging of thick biological specimens is valuable for identifying biologically- or pathologically-relevant features of interest, especially for subsequent correlative studies. Unfortunately, high-resolution imaging techniques at cryogenic conditio...

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Autores principales:	S. H. Shahmoradian, E. H. R. Tsai, A. Diaz, M. Guizar-Sicairos, J. Raabe, L. Spycher, M. Britschgi, A. Ruf, H. Stahlberg, M. Holler
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/10c2cab067824999bee6a862a246bb92
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spelling	oai:doaj.org-article:10c2cab067824999bee6a862a246bb922021-12-02T11:41:22ZThree-Dimensional Imaging of Biological Tissue by Cryo X-Ray Ptychography10.1038/s41598-017-05587-42045-2322https://doaj.org/article/10c2cab067824999bee6a862a246bb922017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05587-4https://doaj.org/toc/2045-2322Abstract High-throughput three-dimensional cryogenic imaging of thick biological specimens is valuable for identifying biologically- or pathologically-relevant features of interest, especially for subsequent correlative studies. Unfortunately, high-resolution imaging techniques at cryogenic conditions often require sample reduction through sequential physical milling or sectioning for sufficient penetration to generate each image of the 3-D stack. This study represents the first demonstration of using ptychographic hard X-ray tomography at cryogenic temperatures for imaging thick biological tissue in a chemically-fixed, frozen-hydrated state without heavy metal staining and organic solvents. Applied to mammalian brain, this label-free cryogenic imaging method allows visualization of myelinated axons and sub-cellular features such as age-related pigmented cellular inclusions at a spatial resolution of ~100 nanometers and thicknesses approaching 100 microns. Because our approach does not require dehydration, staining or reduction of the sample, we introduce the possibility for subsequent analysis of the same tissue using orthogonal approaches that are expected to yield direct complementary insight to the biological features of interest.S. H. ShahmoradianE. H. R. TsaiA. DiazM. Guizar-SicairosJ. RaabeL. SpycherM. BritschgiA. RufH. StahlbergM. HollerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-12 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q S. H. Shahmoradian E. H. R. Tsai A. Diaz M. Guizar-Sicairos J. Raabe L. Spycher M. Britschgi A. Ruf H. Stahlberg M. Holler Three-Dimensional Imaging of Biological Tissue by Cryo X-Ray Ptychography
description	Abstract High-throughput three-dimensional cryogenic imaging of thick biological specimens is valuable for identifying biologically- or pathologically-relevant features of interest, especially for subsequent correlative studies. Unfortunately, high-resolution imaging techniques at cryogenic conditions often require sample reduction through sequential physical milling or sectioning for sufficient penetration to generate each image of the 3-D stack. This study represents the first demonstration of using ptychographic hard X-ray tomography at cryogenic temperatures for imaging thick biological tissue in a chemically-fixed, frozen-hydrated state without heavy metal staining and organic solvents. Applied to mammalian brain, this label-free cryogenic imaging method allows visualization of myelinated axons and sub-cellular features such as age-related pigmented cellular inclusions at a spatial resolution of ~100 nanometers and thicknesses approaching 100 microns. Because our approach does not require dehydration, staining or reduction of the sample, we introduce the possibility for subsequent analysis of the same tissue using orthogonal approaches that are expected to yield direct complementary insight to the biological features of interest.
format	article
author	S. H. Shahmoradian E. H. R. Tsai A. Diaz M. Guizar-Sicairos J. Raabe L. Spycher M. Britschgi A. Ruf H. Stahlberg M. Holler
author_facet	S. H. Shahmoradian E. H. R. Tsai A. Diaz M. Guizar-Sicairos J. Raabe L. Spycher M. Britschgi A. Ruf H. Stahlberg M. Holler
author_sort	S. H. Shahmoradian
title	Three-Dimensional Imaging of Biological Tissue by Cryo X-Ray Ptychography
title_short	Three-Dimensional Imaging of Biological Tissue by Cryo X-Ray Ptychography
title_full	Three-Dimensional Imaging of Biological Tissue by Cryo X-Ray Ptychography
title_fullStr	Three-Dimensional Imaging of Biological Tissue by Cryo X-Ray Ptychography
title_full_unstemmed	Three-Dimensional Imaging of Biological Tissue by Cryo X-Ray Ptychography
title_sort	three-dimensional imaging of biological tissue by cryo x-ray ptychography
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/10c2cab067824999bee6a862a246bb92
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Three-Dimensional Imaging of Biological Tissue by Cryo X-Ray Ptychography

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