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

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...

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Main Authors: Anna E Scott, Dragos M Vasilescu, Katherine A D Seal, Samuel D Keyes, Mark N Mavrogordato, James C Hogg, Ian Sinclair, Jane A Warner, Tillie-Louise Hackett, Peter M Lackie
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2015-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC4452358?pdf=render
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author Anna E Scott
Dragos M Vasilescu
Katherine A D Seal
Samuel D Keyes
Mark N Mavrogordato
James C Hogg
Ian Sinclair
Jane A Warner
Tillie-Louise Hackett
Peter M Lackie
author_facet Anna E Scott
Dragos M Vasilescu
Katherine A D Seal
Samuel D Keyes
Mark N Mavrogordato
James C Hogg
Ian Sinclair
Jane A Warner
Tillie-Louise Hackett
Peter M Lackie
author_sort Anna E Scott
collection DOAJ
description 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.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.The µCT imaging protocol provided contrast between tissue and paraffin in FFPE samples (15 mm x 7 mm). 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.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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spelling doaj.art-25716139b176413e98a361231351d1e22022-12-21T17:30:34ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01106e012623010.1371/journal.pone.0126230Three dimensional imaging of paraffin embedded human lung tissue samples by micro-computed tomography.Anna E ScottDragos M VasilescuKatherine A D SealSamuel D KeyesMark N MavrogordatoJames C HoggIan SinclairJane A WarnerTillie-Louise HackettPeter M LackieUnderstanding 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.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.The µCT imaging protocol provided contrast between tissue and paraffin in FFPE samples (15 mm x 7 mm). 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.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.http://europepmc.org/articles/PMC4452358?pdf=render
spellingShingle Anna E Scott
Dragos M Vasilescu
Katherine A D Seal
Samuel D Keyes
Mark N Mavrogordato
James C Hogg
Ian Sinclair
Jane A Warner
Tillie-Louise Hackett
Peter M Lackie
Three dimensional imaging of paraffin embedded human lung tissue samples by micro-computed tomography.
PLoS ONE
title Three dimensional imaging of paraffin embedded human lung tissue samples by micro-computed tomography.
title_full Three dimensional imaging of paraffin embedded human lung tissue samples by micro-computed tomography.
title_fullStr Three dimensional imaging of paraffin embedded human lung tissue samples by micro-computed tomography.
title_full_unstemmed Three dimensional imaging of paraffin embedded human lung tissue samples by micro-computed tomography.
title_short Three dimensional imaging of paraffin embedded human lung tissue samples by micro-computed tomography.
title_sort three dimensional imaging of paraffin embedded human lung tissue samples by micro computed tomography
url http://europepmc.org/articles/PMC4452358?pdf=render
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