The University of Southampton
University of Southampton Institutional Repository
Warning ePrints Soton is experiencing an issue with some file downloads not being available. We are working hard to fix this. Please bear with us.

Correlative 3D imaging and microfluidic modelling of human pulmonary lymphatics using immunohistochemistry and high-resolution μCT

Correlative 3D imaging and microfluidic modelling of human pulmonary lymphatics using immunohistochemistry and high-resolution μCT
Correlative 3D imaging and microfluidic modelling of human pulmonary lymphatics using immunohistochemistry and high-resolution μCT
Lung lymphatics maintain fluid homoeostasis by providing a drainage system that returns fluid, cells and metabolites to the circulatory system. The 3D structure of the human pulmonary lymphatic network is essential to lung function, but it is poorly characterised. Image-based 3D mathematical modelling of pulmonary lymphatic microfluidics has been limited by the lack of accurate and representative image geometries. This is due to the microstructural similarity of the lymphatics to the blood vessel network, the lack of lymphatic-specific biomarkers, the technical limitations associated with image resolution in 3D, and sectioning artefacts present in 2D techniques. We present a method that combines lymphatic specific (D240 antibody) immunohistochemistry (IHC), optimised high-resolution X-ray microfocus computed tomography (μCT) and finite-element mathematical modelling to assess the function of human peripheral lung tissue. The initial results identify lymphatic heterogeneity within and between lung tissue. Lymphatic vessel volume fraction and fractal dimension significantly decreases away from the lung pleural surface (p<0.001, n=25 and p <0.01, n=20, respectively). Microfluidic modelling successfully shows that in lung tissue the fluid derived from the blood vessels drains through the interstitium into the lymphatic vessel network and this drainage is different in the subpleural space compared to the intralobular space. When comparing lung tissue from health and disease, human pulmonary lymphatics were significantly different across five morphometric measures used in this study (p≤0.0001). This proof of principle study establishes a new engineering technology and workflow for further studies of pulmonary lymphatics and demonstrates for the first time the combination of correlative μCT and IHC to enable 3D mathematical modelling of human lung microfluidics at micrometre resolution.
2045-2322
Robinson, Stephanie
55ce53a2-2ab9-499d-aa59-e8dc8374823e
Ramsden, Jonathan J.
979fa39c-1805-4e05-a266-4a3661fbf440
Warner, Jane
8571b049-31bb-4a2a-a3c7-4184be20fe25
Lackie, Peter
4afbbe1a-22a6-4ceb-8cad-f3696dc43a7a
Roose, Tiina
3581ab5b-71e1-4897-8d88-59f13f3bccfe
Robinson, Stephanie
55ce53a2-2ab9-499d-aa59-e8dc8374823e
Ramsden, Jonathan J.
979fa39c-1805-4e05-a266-4a3661fbf440
Warner, Jane
8571b049-31bb-4a2a-a3c7-4184be20fe25
Lackie, Peter
4afbbe1a-22a6-4ceb-8cad-f3696dc43a7a
Roose, Tiina
3581ab5b-71e1-4897-8d88-59f13f3bccfe

Robinson, Stephanie, Ramsden, Jonathan J., Warner, Jane, Lackie, Peter and Roose, Tiina (2019) Correlative 3D imaging and microfluidic modelling of human pulmonary lymphatics using immunohistochemistry and high-resolution μCT. Scientific Reports, 9, [6415]. (doi:10.1038/s41598-019-42794-7).

Record type: Article

Abstract

Lung lymphatics maintain fluid homoeostasis by providing a drainage system that returns fluid, cells and metabolites to the circulatory system. The 3D structure of the human pulmonary lymphatic network is essential to lung function, but it is poorly characterised. Image-based 3D mathematical modelling of pulmonary lymphatic microfluidics has been limited by the lack of accurate and representative image geometries. This is due to the microstructural similarity of the lymphatics to the blood vessel network, the lack of lymphatic-specific biomarkers, the technical limitations associated with image resolution in 3D, and sectioning artefacts present in 2D techniques. We present a method that combines lymphatic specific (D240 antibody) immunohistochemistry (IHC), optimised high-resolution X-ray microfocus computed tomography (μCT) and finite-element mathematical modelling to assess the function of human peripheral lung tissue. The initial results identify lymphatic heterogeneity within and between lung tissue. Lymphatic vessel volume fraction and fractal dimension significantly decreases away from the lung pleural surface (p<0.001, n=25 and p <0.01, n=20, respectively). Microfluidic modelling successfully shows that in lung tissue the fluid derived from the blood vessels drains through the interstitium into the lymphatic vessel network and this drainage is different in the subpleural space compared to the intralobular space. When comparing lung tissue from health and disease, human pulmonary lymphatics were significantly different across five morphometric measures used in this study (p≤0.0001). This proof of principle study establishes a new engineering technology and workflow for further studies of pulmonary lymphatics and demonstrates for the first time the combination of correlative μCT and IHC to enable 3D mathematical modelling of human lung microfluidics at micrometre resolution.

Text
Main document resubmission - Accepted Manuscript
Available under License Creative Commons Attribution.
Download (100kB)
Text
s41598-019-42794-7 - Version of Record
Available under License Creative Commons Attribution.
Download (3MB)

More information

Submitted date: 2017
Accepted/In Press date: 8 April 2019
e-pub ahead of print date: 23 April 2019

Identifiers

Local EPrints ID: 418311
URI: http://eprints.soton.ac.uk/id/eprint/418311
ISSN: 2045-2322
PURE UUID: 53df9ce9-bea9-4016-aaa0-cd40b4b9532b
ORCID for Stephanie Robinson: ORCID iD orcid.org/0000-0001-5436-2929
ORCID for Peter Lackie: ORCID iD orcid.org/0000-0001-7138-3764
ORCID for Tiina Roose: ORCID iD orcid.org/0000-0001-8710-1063

Catalogue record

Date deposited: 27 Feb 2018 17:32
Last modified: 22 Nov 2021 03:12

Export record

Altmetrics

Contributors

Author: Stephanie Robinson ORCID iD
Author: Jonathan J. Ramsden
Author: Jane Warner
Author: Peter Lackie ORCID iD
Author: Tiina Roose ORCID iD

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of http://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×