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Three dimensional imaging of paraffin embedded human lung tissue samples by micro-computed tomography

Three dimensional imaging of paraffin embedded human lung tissue samples by micro-computed tomography
Three dimensional imaging of paraffin embedded human lung tissue samples by micro-computed tomography
Background: understanding the three-dimensional (3-D) micro-architecture of lung tissue can provide insights into the pathology of lung disease. Micro computed tomography (µCT) has previously been used to elucidate lung 3D histology and morphometry in fixed samples that have been stained with contrast agents or air inflated and dried. However, non-destructive microstructural 3D imaging of formalin-fixed paraffin embedded (FFPE) tissues would facilitate retrospective analysis of extensive tissue archives of lung FFPE lung samples with linked clinical data.

Methods: FFPE human lung tissue samples (n = 4) were scanned using a Nikon metrology µCT scanner. Semi-automatic techniques were used to segment the 3D structure of airways and blood vessels. Airspace size (mean linear intercept, Lm) was measured on µCT images and on matched histological sections from the same FFPE samples imaged by light microscopy to validate µCT imaging.

Results: the µCT imaging protocol provided contrast between tissue and paraffin in FFPE samples (15mm x 7mm). Resolution (voxel size 6.7 µm) in the reconstructed images was sufficient for semi-automatic image segmentation of airways and blood vessels as well as quantitative airspace analysis. The scans were also used to scout for regions of interest, enabling time-efficient preparation of conventional histological sections. The Lm measurements from µCT images were not significantly different to those from matched histological sections.

Conclusion: we demonstrated how non-destructive imaging of routinely prepared FFPE samples by laboratory µCT can be used to visualize and assess the 3D morphology of the lung including by morphometric analysis.
1932-6203
1-10
Scott, A.E.
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Vasilescu, D.M.
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Seal, K.A.D.
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Keyes, S.D.
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Mavrogordato, M.N.
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Hogg, J.C.
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Sinclair, I.
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Warner, J.A.
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Hackett, T.L.
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Lackie, P.M.
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Scott, A.E.
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Vasilescu, D.M.
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Seal, K.A.D.
9a6fe730-8b7b-42cc-bba5-eba396f392f4
Keyes, S.D.
c7c08bbb-8202-4101-97de-548d3585c368
Mavrogordato, M.N.
f3e0879b-118a-463a-a130-1c890e9ab547
Hogg, J.C.
709e6724-1c15-4277-8e34-b945b99ae7ca
Sinclair, I.
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Warner, J.A.
8571b049-31bb-4a2a-a3c7-4184be20fe25
Hackett, T.L.
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Lackie, P.M.
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Scott, A.E., Vasilescu, D.M. and Seal, K.A.D. et al. (2015) Three dimensional imaging of paraffin embedded human lung tissue samples by micro-computed tomography. PLoS ONE, 1-10. (doi:10.1371/journal.pone.0126230).

Record type: Article

Abstract

Background: understanding the three-dimensional (3-D) micro-architecture of lung tissue can provide insights into the pathology of lung disease. Micro computed tomography (µCT) has previously been used to elucidate lung 3D histology and morphometry in fixed samples that have been stained with contrast agents or air inflated and dried. However, non-destructive microstructural 3D imaging of formalin-fixed paraffin embedded (FFPE) tissues would facilitate retrospective analysis of extensive tissue archives of lung FFPE lung samples with linked clinical data.

Methods: FFPE human lung tissue samples (n = 4) were scanned using a Nikon metrology µCT scanner. Semi-automatic techniques were used to segment the 3D structure of airways and blood vessels. Airspace size (mean linear intercept, Lm) was measured on µCT images and on matched histological sections from the same FFPE samples imaged by light microscopy to validate µCT imaging.

Results: the µCT imaging protocol provided contrast between tissue and paraffin in FFPE samples (15mm x 7mm). Resolution (voxel size 6.7 µm) in the reconstructed images was sufficient for semi-automatic image segmentation of airways and blood vessels as well as quantitative airspace analysis. The scans were also used to scout for regions of interest, enabling time-efficient preparation of conventional histological sections. The Lm measurements from µCT images were not significantly different to those from matched histological sections.

Conclusion: we demonstrated how non-destructive imaging of routinely prepared FFPE samples by laboratory µCT can be used to visualize and assess the 3D morphology of the lung including by morphometric analysis.

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More information

Accepted/In Press date: 30 March 2015
Published date: 1 June 2015
Additional Information: A first demonstration of diagnostic potential of laboratory µCT in clinically-representative tissue blocks, collaborating with world-leading Centre (UBC, Vancouver). We have subsequently evidenced such data can overturn erroneous understanding of disease pathogenesis, shifting clinical histopathology from 150 years of 2D practice, into 3D. Subsequent Wellcome Trust support has facilitated development of a new generation of clinically driven µCT machines with Nikon UK: first unit (“Med-X”) has been deployed at Southampton General Hospital (August 2016) and follow-on units are shipping worldwide. A pre-clinical trial has since been carried out, drawing together a panel of world-class pathologists (USA, UK, Italy, Japan).
Organisations: Faculty of Medicine, Faculty of Engineering and the Environment

Identifiers

Local EPrints ID: 381745
URI: http://eprints.soton.ac.uk/id/eprint/381745
ISSN: 1932-6203
PURE UUID: 1a31b6d9-5e62-4a2c-b21c-6026e0110680
ORCID for P.M. Lackie: ORCID iD orcid.org/0000-0001-7138-3764

Catalogue record

Date deposited: 12 Oct 2015 13:24
Last modified: 17 Dec 2019 02:00

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