Journal
JOURNAL OF APPLIED PHYSIOLOGY
Volume 122, Issue 1, Pages 161-169Publisher
AMER PHYSIOLOGICAL SOC
DOI: 10.1152/japplphysiol.00838.2016
Keywords
cryostage; gene expression; lung; micro-computed tomography; soft tissue imaging
Categories
Funding
- British Columbia Lung Association
- Alpha-1 Foundation
- IMPACT-Canadian Institutes of Health Research fellowship program
- NATIONAL HEART, LUNG, AND BLOOD INSTITUTE [R01HL118542] Funding Source: NIH RePORTER
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Micro-computed tomography (CT) enables three-dimensional (3D) imaging of complex soft tissue structures, but current protocols used to achieve this goal preclude cellular and molecular phenotyping of the tissue. Here we describe a radiolucent cryostage that permits micro-CT imaging of unfixed frozen human lung samples at an isotropic voxel size of (11 mu m)(3) under conditions where the sample is maintained frozen at - 30 degrees C during imaging. The cryostage was tested for thermal stability to maintain samples frozen up to 8 h. This report describes the methods used to choose the materials required for cryostage construction and demonstrates that whole genome mRNA integrity and expression are not compromised by exposure to micro-CT radiation and that the tissue can be used for immunohistochemistry. The new cryostage provides a novel method enabling integration of 3D tissue structure with cellular and molecular analysis to facilitate the identification of molecular determinants of disease.
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