Format

Send to

Choose Destination
Sci Rep. 2017 Jul 24;7(1):6291. doi: 10.1038/s41598-017-05587-4.

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

Author information

1
Paul Scherrer Institut, Laboratory for Biomolecular Research, Department of Biology and Chemistry, 5232, Villigen PSI, Switzerland. sarah.shahmoradian@psi.ch.
2
Paul Scherrer Institut, Laboratory for Macromolecules and Bioimaging, Department of Synchrotron Radiation and Nanotechnology, 5232, Villigen PSI, Switzerland.
3
Paul Scherrer Institut, Laboratory for Synchrotron Radiation Condensed Matter, Department of Synchrotron Radiation and Nanotechnology, 5232, Villigen PSI, Switzerland.
4
Roche Pharma Research and Early Development, NORD DTA, Roche Innovation Center Basel, 4070, Basel, Switzerland.
5
Roche Pharma Research and Early Development, Chemical Biology, Roche Innovation Center Basel, 4070, Basel, Switzerland.
6
Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, 4056, Basel, Switzerland.

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.

PMID:
28740127
PMCID:
PMC5524705
DOI:
10.1038/s41598-017-05587-4
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center