Light sheet fluorescence microscopy of cleared human eyes

We provide here a procedure enabling light sheet fluorescence microscopy (LSFM) of entire human eyes after iDISCO + -based clearing (ClearEye) and immunolabeling. Demonstrated here in four eyes, post-processing of LSFM stacks enables three-dimensional (3D) navigation and customized display, includin...

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Autores principales: Darche, Marie, Borella, Ysé, Verschueren, Anna, Gantar, Ivana, Pagès, Stéphane, Batti, Laura, Paques, Michel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10564773/
https://www.ncbi.nlm.nih.gov/pubmed/37816868
http://dx.doi.org/10.1038/s42003-023-05401-0
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author Darche, Marie
Borella, Ysé
Verschueren, Anna
Gantar, Ivana
Pagès, Stéphane
Batti, Laura
Paques, Michel
author_facet Darche, Marie
Borella, Ysé
Verschueren, Anna
Gantar, Ivana
Pagès, Stéphane
Batti, Laura
Paques, Michel
author_sort Darche, Marie
collection PubMed
description We provide here a procedure enabling light sheet fluorescence microscopy (LSFM) of entire human eyes after iDISCO + -based clearing (ClearEye) and immunolabeling. Demonstrated here in four eyes, post-processing of LSFM stacks enables three-dimensional (3D) navigation and customized display, including en face viewing of the fundus similarly to clinical imaging, with resolution of retinal capillaries. This method overcomes several limitations of traditional histology of the eyes. Tracing of spatially complex structures such as anterior ciliary vessels and Schlemm’s canal was achieved. We conclude that LSFM of immunolabeled human eyes after iDISCO + -based clearing is a powerful tool for 3D histology of large human ocular samples, including entire eyes, which will be useful in both anatomopathology and in research.
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spelling pubmed-105647732023-10-12 Light sheet fluorescence microscopy of cleared human eyes Darche, Marie Borella, Ysé Verschueren, Anna Gantar, Ivana Pagès, Stéphane Batti, Laura Paques, Michel Commun Biol Article We provide here a procedure enabling light sheet fluorescence microscopy (LSFM) of entire human eyes after iDISCO + -based clearing (ClearEye) and immunolabeling. Demonstrated here in four eyes, post-processing of LSFM stacks enables three-dimensional (3D) navigation and customized display, including en face viewing of the fundus similarly to clinical imaging, with resolution of retinal capillaries. This method overcomes several limitations of traditional histology of the eyes. Tracing of spatially complex structures such as anterior ciliary vessels and Schlemm’s canal was achieved. We conclude that LSFM of immunolabeled human eyes after iDISCO + -based clearing is a powerful tool for 3D histology of large human ocular samples, including entire eyes, which will be useful in both anatomopathology and in research. Nature Publishing Group UK 2023-10-10 /pmc/articles/PMC10564773/ /pubmed/37816868 http://dx.doi.org/10.1038/s42003-023-05401-0 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Darche, Marie
Borella, Ysé
Verschueren, Anna
Gantar, Ivana
Pagès, Stéphane
Batti, Laura
Paques, Michel
Light sheet fluorescence microscopy of cleared human eyes
title Light sheet fluorescence microscopy of cleared human eyes
title_full Light sheet fluorescence microscopy of cleared human eyes
title_fullStr Light sheet fluorescence microscopy of cleared human eyes
title_full_unstemmed Light sheet fluorescence microscopy of cleared human eyes
title_short Light sheet fluorescence microscopy of cleared human eyes
title_sort light sheet fluorescence microscopy of cleared human eyes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10564773/
https://www.ncbi.nlm.nih.gov/pubmed/37816868
http://dx.doi.org/10.1038/s42003-023-05401-0
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