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Multiscale and Multimodal Optical Imaging of the Ultrastructure of Human Liver Biopsies

The liver as the largest organ in the human body is composed of a complex macroscopic and microscopic architecture that supports its indispensable function to maintain physiological homeostasis. Optical imaging of the human liver is particularly challenging because of the need to cover length scales...

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Autores principales: Kong, Cihang, Bobe, Stefanie, Pilger, Christian, Lachetta, Mario, Øie, Cristina Ionica, Kirschnick, Nils, Mönkemöller, Viola, Hübner, Wolfgang, Förster, Christine, Schüttpelz, Mark, Kiefer, Friedemann, Huser, Thomas, Schulte am Esch, Jan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7925637/
https://www.ncbi.nlm.nih.gov/pubmed/33679449
http://dx.doi.org/10.3389/fphys.2021.637136
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author Kong, Cihang
Bobe, Stefanie
Pilger, Christian
Lachetta, Mario
Øie, Cristina Ionica
Kirschnick, Nils
Mönkemöller, Viola
Hübner, Wolfgang
Förster, Christine
Schüttpelz, Mark
Kiefer, Friedemann
Huser, Thomas
Schulte am Esch, Jan
author_facet Kong, Cihang
Bobe, Stefanie
Pilger, Christian
Lachetta, Mario
Øie, Cristina Ionica
Kirschnick, Nils
Mönkemöller, Viola
Hübner, Wolfgang
Förster, Christine
Schüttpelz, Mark
Kiefer, Friedemann
Huser, Thomas
Schulte am Esch, Jan
author_sort Kong, Cihang
collection PubMed
description The liver as the largest organ in the human body is composed of a complex macroscopic and microscopic architecture that supports its indispensable function to maintain physiological homeostasis. Optical imaging of the human liver is particularly challenging because of the need to cover length scales across 7 orders of magnitude (from the centimeter scale to the nanometer scale) in order to fully assess the ultrastructure of the entire organ down to the subcellular scale and probe its physiological function. This task becomes even more challenging the deeper within the organ one hopes to image, because of the strong absorption and scattering of visible light by the liver. Here, we demonstrate how optical imaging methods utilizing highly specific fluorescent labels, as well as label-free optical methods can seamlessly cover this entire size range in excised, fixed human liver tissue and we exemplify this by reconstructing the biliary tree in three-dimensional space. Imaging of tissue beyond approximately 0.5 mm length requires optical clearing of the human liver. We present the successful use of optical projection tomography and light-sheet fluorescence microscopy to derive information about the liver architecture on the millimeter scale. The intermediate size range is covered using label-free structural and chemically sensitive methods, such as second harmonic generation and coherent anti-Stokes Raman scattering microscopy. Laser-scanning confocal microscopy extends the resolution to the nanoscale, allowing us to ultimately image individual liver sinusoidal endothelial cells and their fenestrations by super-resolution structured illumination microscopy. This allowed us to visualize the human hepatobiliary system in 3D down to the cellular level, which indicates that reticular biliary networks communicate with portal bile ducts via single or a few ductuli. Non-linear optical microscopy enabled us to identify fibrotic regions extending from the portal field to the parenchyma, along with microvesicular steatosis in liver biopsies from an older patient. Lastly, super-resolution microscopy allowed us to visualize and determine the size distribution of fenestrations in human liver sinusoidal endothelial cells for the first time under aqueous conditions. Thus, this proof-of-concept study allows us to demonstrate, how, in combination, these techniques open up a new chapter in liver biopsy analysis.
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spelling pubmed-79256372021-03-04 Multiscale and Multimodal Optical Imaging of the Ultrastructure of Human Liver Biopsies Kong, Cihang Bobe, Stefanie Pilger, Christian Lachetta, Mario Øie, Cristina Ionica Kirschnick, Nils Mönkemöller, Viola Hübner, Wolfgang Förster, Christine Schüttpelz, Mark Kiefer, Friedemann Huser, Thomas Schulte am Esch, Jan Front Physiol Physiology The liver as the largest organ in the human body is composed of a complex macroscopic and microscopic architecture that supports its indispensable function to maintain physiological homeostasis. Optical imaging of the human liver is particularly challenging because of the need to cover length scales across 7 orders of magnitude (from the centimeter scale to the nanometer scale) in order to fully assess the ultrastructure of the entire organ down to the subcellular scale and probe its physiological function. This task becomes even more challenging the deeper within the organ one hopes to image, because of the strong absorption and scattering of visible light by the liver. Here, we demonstrate how optical imaging methods utilizing highly specific fluorescent labels, as well as label-free optical methods can seamlessly cover this entire size range in excised, fixed human liver tissue and we exemplify this by reconstructing the biliary tree in three-dimensional space. Imaging of tissue beyond approximately 0.5 mm length requires optical clearing of the human liver. We present the successful use of optical projection tomography and light-sheet fluorescence microscopy to derive information about the liver architecture on the millimeter scale. The intermediate size range is covered using label-free structural and chemically sensitive methods, such as second harmonic generation and coherent anti-Stokes Raman scattering microscopy. Laser-scanning confocal microscopy extends the resolution to the nanoscale, allowing us to ultimately image individual liver sinusoidal endothelial cells and their fenestrations by super-resolution structured illumination microscopy. This allowed us to visualize the human hepatobiliary system in 3D down to the cellular level, which indicates that reticular biliary networks communicate with portal bile ducts via single or a few ductuli. Non-linear optical microscopy enabled us to identify fibrotic regions extending from the portal field to the parenchyma, along with microvesicular steatosis in liver biopsies from an older patient. Lastly, super-resolution microscopy allowed us to visualize and determine the size distribution of fenestrations in human liver sinusoidal endothelial cells for the first time under aqueous conditions. Thus, this proof-of-concept study allows us to demonstrate, how, in combination, these techniques open up a new chapter in liver biopsy analysis. Frontiers Media S.A. 2021-02-17 /pmc/articles/PMC7925637/ /pubmed/33679449 http://dx.doi.org/10.3389/fphys.2021.637136 Text en Copyright © 2021 Kong, Bobe, Pilger, Lachetta, Øie, Kirschnick, Mönkemöller, Hübner, Förster, Schüttpelz, Kiefer, Huser and Schulte am Esch. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Physiology
Kong, Cihang
Bobe, Stefanie
Pilger, Christian
Lachetta, Mario
Øie, Cristina Ionica
Kirschnick, Nils
Mönkemöller, Viola
Hübner, Wolfgang
Förster, Christine
Schüttpelz, Mark
Kiefer, Friedemann
Huser, Thomas
Schulte am Esch, Jan
Multiscale and Multimodal Optical Imaging of the Ultrastructure of Human Liver Biopsies
title Multiscale and Multimodal Optical Imaging of the Ultrastructure of Human Liver Biopsies
title_full Multiscale and Multimodal Optical Imaging of the Ultrastructure of Human Liver Biopsies
title_fullStr Multiscale and Multimodal Optical Imaging of the Ultrastructure of Human Liver Biopsies
title_full_unstemmed Multiscale and Multimodal Optical Imaging of the Ultrastructure of Human Liver Biopsies
title_short Multiscale and Multimodal Optical Imaging of the Ultrastructure of Human Liver Biopsies
title_sort multiscale and multimodal optical imaging of the ultrastructure of human liver biopsies
topic Physiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7925637/
https://www.ncbi.nlm.nih.gov/pubmed/33679449
http://dx.doi.org/10.3389/fphys.2021.637136
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