Correlative 3D Imaging and Microfluidic Modelling of Human Pulmonary Lymphatics using Immunohistochemistry and High-resolution μCT

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 mathematica...

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Autores principales:	Stephanie K. Robinson, Jonathan J. Ramsden, Jane Warner, Peter M. Lackie, Tiina Roose
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2019
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b58fc3cc5e714adfbe6a2de50ebebe48
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spelling	oai:doaj.org-article:b58fc3cc5e714adfbe6a2de50ebebe482021-12-02T15:07:53ZCorrelative 3D Imaging and Microfluidic Modelling of Human Pulmonary Lymphatics using Immunohistochemistry and High-resolution μCT10.1038/s41598-019-42794-72045-2322https://doaj.org/article/b58fc3cc5e714adfbe6a2de50ebebe482019-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-42794-7https://doaj.org/toc/2045-2322Abstract 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.Stephanie K. RobinsonJonathan J. RamsdenJane WarnerPeter M. LackieTiina RooseNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-14 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Stephanie K. Robinson Jonathan J. Ramsden Jane Warner Peter M. Lackie Tiina Roose Correlative 3D Imaging and Microfluidic Modelling of Human Pulmonary Lymphatics using Immunohistochemistry and High-resolution μCT
description	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.
format	article
author	Stephanie K. Robinson Jonathan J. Ramsden Jane Warner Peter M. Lackie Tiina Roose
author_facet	Stephanie K. Robinson Jonathan J. Ramsden Jane Warner Peter M. Lackie Tiina Roose
author_sort	Stephanie K. Robinson
title	Correlative 3D Imaging and Microfluidic Modelling of Human Pulmonary Lymphatics using Immunohistochemistry and High-resolution μCT
title_short	Correlative 3D Imaging and Microfluidic Modelling of Human Pulmonary Lymphatics using Immunohistochemistry and High-resolution μCT
title_full	Correlative 3D Imaging and Microfluidic Modelling of Human Pulmonary Lymphatics using Immunohistochemistry and High-resolution μCT
title_fullStr	Correlative 3D Imaging and Microfluidic Modelling of Human Pulmonary Lymphatics using Immunohistochemistry and High-resolution μCT
title_full_unstemmed	Correlative 3D Imaging and Microfluidic Modelling of Human Pulmonary Lymphatics using Immunohistochemistry and High-resolution μCT
title_sort	correlative 3d imaging and microfluidic modelling of human pulmonary lymphatics using immunohistochemistry and high-resolution μct
publisher	Nature Portfolio
publishDate	2019
url	https://doaj.org/article/b58fc3cc5e714adfbe6a2de50ebebe48
work_keys_str_mv	AT stephaniekrobinson correlative3dimagingandmicrofluidicmodellingofhumanpulmonarylymphaticsusingimmunohistochemistryandhighresolutionmct AT jonathanjramsden correlative3dimagingandmicrofluidicmodellingofhumanpulmonarylymphaticsusingimmunohistochemistryandhighresolutionmct AT janewarner correlative3dimagingandmicrofluidicmodellingofhumanpulmonarylymphaticsusingimmunohistochemistryandhighresolutionmct AT petermlackie correlative3dimagingandmicrofluidicmodellingofhumanpulmonarylymphaticsusingimmunohistochemistryandhighresolutionmct AT tiinaroose correlative3dimagingandmicrofluidicmodellingofhumanpulmonarylymphaticsusingimmunohistochemistryandhighresolutionmct
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Correlative 3D Imaging and Microfluidic Modelling of Human Pulmonary Lymphatics using Immunohistochemistry and High-resolution μCT

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