title: Nondestructive cryomicro-CT imaging enables structural and molecular analysis of human lung tissue.
abstract: 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 µm)3under conditions where the sample is maintained frozen at -30°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.
  
  
  
  The described micro-CT cryostage provides a novel way to study the three-dimensional lung structure preserved without the effects of fixatives while enabling subsequent studies of the cellular matrix composition and gene expression. This approach will, for the first time, enable researchers to study structural changes of lung tissues that occur with disease and correlate them with changes in gene or protein signatures.
authors.affiliation: University of British Columbia, Centre for Heart Lung Innovation, Vancouver, British Columbia, Canada; dragos.vasilescu@hli.ubc.ca.
authors.forename: Dragoş M
authors.initials: DM
authors.lastname: Vasilescu
authors.affiliation: Materials Science and Engineering, McMaster University, Hamilton, Ontario, Canada.
authors.forename: André B
authors.initials: AB
authors.lastname: Phillion
authors.affiliation: University of British Columbia, Centre for Heart Lung Innovation, Vancouver, British Columbia, Canada.
authors.forename: Naoya
authors.initials: N
authors.lastname: Tanabe
authors.affiliation: University of British Columbia, Centre for Heart Lung Innovation, Vancouver, British Columbia, Canada.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Kinose
authors.affiliation: Paige Instruments, Woodland, California.
authors.forename: David F
authors.initials: DF
authors.lastname: Paige
authors.affiliation: Section of Computational Biomedicine, Department of Medicine, Boston University Medical Center, Boston, Massachusetts.
authors.forename: Jacob J
authors.initials: JJ
authors.lastname: Kantrowitz
authors.affiliation: Section of Computational Biomedicine, Department of Medicine, Boston University Medical Center, Boston, Massachusetts.
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
authors.affiliation: Section of Computational Biomedicine, Department of Medicine, Boston University Medical Center, Boston, Massachusetts.
authors.forename: Hanqiao
authors.initials: H
authors.lastname: Liu
authors.affiliation: University of British Columbia, Centre for Heart Lung Innovation, Vancouver, British Columbia, Canada.
authors.forename: Nick
authors.initials: N
authors.lastname: Fishbane
authors.affiliation: Lung Transplant Unit, Department of Clinical and Experimental Medicine, KU Leuven, Leuven, Belgium.
authors.forename: Stijn E
authors.initials: SE
authors.lastname: Verleden
authors.affiliation: Lung Transplant Unit, Department of Clinical and Experimental Medicine, KU Leuven, Leuven, Belgium.
authors.forename: Bart M
authors.initials: BM
authors.lastname: Vanaudenaerde
authors.affiliation: Section of Computational Biomedicine, Department of Medicine, Boston University Medical Center, Boston, Massachusetts.
authors.forename: Marc
authors.initials: M
authors.lastname: Lenburg
authors.affiliation: Janssen Disease Interception Accelerator, Janssen Pharmaceutical Companies of Johnson and Johnson, Raritan, New Jersey; and.
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Stevenson
authors.affiliation: Section of Computational Biomedicine, Department of Medicine, Boston University Medical Center, Boston, Massachusetts.
authors.forename: Avrum
authors.initials: A
authors.lastname: Spira
authors.affiliation: Division of Thoracic Surgery, University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Joel D
authors.initials: JD
authors.lastname: Cooper
authors.affiliation: University of British Columbia, Centre for Heart Lung Innovation, Vancouver, British Columbia, Canada.
authors.forename: Tillie-Louise
authors.initials: TL
authors.lastname: Hackett
authors.affiliation: University of British Columbia, Centre for Heart Lung Innovation, Vancouver, British Columbia, Canada.
authors.forename: James C
authors.initials: JC
authors.lastname: Hogg
mesh_terms: D015870:Gene Expression / Q000502:physiology; D016678:Genome / Q000502:physiology; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods; D008168:Lung / Q000378:metabolism / Q000473:pathology*; D012333:RNA, Messenger / Q000378:metabolism; D055114:X-Ray Microtomography / Q000379:methods*
chemical_list: D012333:RNA, Messenger
keywords: cryostage; gene expression; lung; micro-computed tomography; soft tissue imaging