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3D mapping of blood vessel networks and cells in COPD and non-COPD lung tissue samples using micro-computed tomography and immunofluorescence

3D mapping of blood vessel networks and cells in COPD and non-COPD lung tissue samples using micro-computed tomography and immunofluorescence
3D mapping of blood vessel networks and cells in COPD and non-COPD lung tissue samples using micro-computed tomography and immunofluorescence
Micro-computed tomography (µCT) is a non-destructive 3D imaging technique used to map tissue microstructure at typical resolutions of 1-10 µm. Correlated with 2D immunofluorescence (IF) microscopy pathophysiologically relevant cells can be identified in 3D. We aimed to investigate 3D networks of small blood vessels (<2mm) and the distribution of immune cells in mild-moderate COPD patients. FFPE peripheral lung samples from 5 non-COPD and 5 COPD patients were scanned using µCT. IF staining for smooth muscle actin, airway epithelium, mast cells and macrophages was digitised, and co-registered with 3D µCT scans. Blood vessels, airways and infiltrating cells were identified semi-automatically by IF in the µCT volume (Fig 1). Quantitative estimates of blood vessel thickness were made, initial modal thickness values were lower in non-COPD (12-25µm) compared to COPD samples (40-60µm). Thousands of macrophages (2100-7200) and mast cells (1700-9000) were localised by IF per tissue section with fewer mast cells (<1500) detected in COPD tissue sections. Combined with the 3D vasculature network the distribution of macrophages and mast cells were found to be similar in both COPD and non-COPD with 70-80% of cells within 2mm of a blood vessel. In summary, we demonstrate an approach to identify, localise and analyse lung networks in 3D and relevant infiltrating cells in human lung disease.`
2312-0541
Lawson, M
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Katsamenis, O
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Olding, M
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Larkin, O
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Smit, B
31968f76-81f2-4560-b5b7-17d8b6925c33
Haig, I
8c3dc208-c92b-4229-ab67-08dda8b45d64
Schneider, P
a810f925-4808-44e4-8a4a-a51586f9d7ad
Lackie, P
4afbbe1a-22a6-4ceb-8cad-f3696dc43a7a
Warner, J
8571b049-31bb-4a2a-a3c7-4184be20fe25
Lawson, M
5c14101b-e305-463d-97fd-d38b0000c3d6
Katsamenis, O
8553e7c3-d860-4b7a-a883-abf6c0c4b438
Olding, M
4194199f-96d5-4c34-90d5-bab40adf06a3
Larkin, O
5bde48ce-ef5d-43fa-998d-f38800240514
Smit, B
31968f76-81f2-4560-b5b7-17d8b6925c33
Haig, I
8c3dc208-c92b-4229-ab67-08dda8b45d64
Schneider, P
a810f925-4808-44e4-8a4a-a51586f9d7ad
Lackie, P
4afbbe1a-22a6-4ceb-8cad-f3696dc43a7a
Warner, J
8571b049-31bb-4a2a-a3c7-4184be20fe25

Lawson, M, Katsamenis, O, Olding, M, Larkin, O, Smit, B, Haig, I, Schneider, P, Lackie, P and Warner, J (2020) 3D mapping of blood vessel networks and cells in COPD and non-COPD lung tissue samples using micro-computed tomography and immunofluorescence. ERJ Open Research, 6 (5). (doi:10.1183/23120541.lsc-2020.21).

Record type: Meeting abstract

Abstract

Micro-computed tomography (µCT) is a non-destructive 3D imaging technique used to map tissue microstructure at typical resolutions of 1-10 µm. Correlated with 2D immunofluorescence (IF) microscopy pathophysiologically relevant cells can be identified in 3D. We aimed to investigate 3D networks of small blood vessels (<2mm) and the distribution of immune cells in mild-moderate COPD patients. FFPE peripheral lung samples from 5 non-COPD and 5 COPD patients were scanned using µCT. IF staining for smooth muscle actin, airway epithelium, mast cells and macrophages was digitised, and co-registered with 3D µCT scans. Blood vessels, airways and infiltrating cells were identified semi-automatically by IF in the µCT volume (Fig 1). Quantitative estimates of blood vessel thickness were made, initial modal thickness values were lower in non-COPD (12-25µm) compared to COPD samples (40-60µm). Thousands of macrophages (2100-7200) and mast cells (1700-9000) were localised by IF per tissue section with fewer mast cells (<1500) detected in COPD tissue sections. Combined with the 3D vasculature network the distribution of macrophages and mast cells were found to be similar in both COPD and non-COPD with 70-80% of cells within 2mm of a blood vessel. In summary, we demonstrate an approach to identify, localise and analyse lung networks in 3D and relevant infiltrating cells in human lung disease.`

Text
ERS Lung Science Conference poster: Lawson et al.
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More information

Accepted/In Press date: 5 March 2020
e-pub ahead of print date: 15 April 2020
Venue - Dates: 18th Lung Science Conference of the European Respiratory Society, , Estoril, Portugal, 2018-03-05 - 2020-03-08

Identifiers

Local EPrints ID: 443734
URI: http://eprints.soton.ac.uk/id/eprint/443734
ISSN: 2312-0541
PURE UUID: bdb65443-4886-425e-8d2d-6d8c16b4544e
ORCID for M Lawson: ORCID iD orcid.org/0000-0003-0115-1698
ORCID for O Katsamenis: ORCID iD orcid.org/0000-0003-4367-4147
ORCID for P Schneider: ORCID iD orcid.org/0000-0001-7499-3576
ORCID for P Lackie: ORCID iD orcid.org/0000-0001-7138-3764

Catalogue record

Date deposited: 10 Sep 2020 16:46
Last modified: 30 Mar 2021 01:54

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Contributors

Author: M Lawson ORCID iD
Author: O Katsamenis ORCID iD
Author: M Olding
Author: O Larkin
Author: B Smit
Author: I Haig
Author: P Schneider ORCID iD
Author: P Lackie ORCID iD
Author: J Warner

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