Hyperglycemia-induced abnormalities in rat and human corneas: the potential of second harmonic generation microscopy.

<h4>Background</h4>Second Harmonic Generation (SHG) microscopy recently appeared as an efficient optical imaging technique to probe unstained collagen-rich tissues like cornea. Moreover, corneal remodeling occurs in many diseases and precise characterization requires overcoming the limit...

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Autores principales:	Gaël Latour, Laura Kowalczuk, Michèle Savoldelli, Jean-Louis Bourges, Karsten Plamann, Francine Behar-Cohen, Marie-Claire Schanne-Klein
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e26093bb65464e42a9b464c1ea46c1bf
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spelling	oai:doaj.org-article:e26093bb65464e42a9b464c1ea46c1bf2021-11-18T08:10:05ZHyperglycemia-induced abnormalities in rat and human corneas: the potential of second harmonic generation microscopy.1932-620310.1371/journal.pone.0048388https://doaj.org/article/e26093bb65464e42a9b464c1ea46c1bf2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23139780/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Second Harmonic Generation (SHG) microscopy recently appeared as an efficient optical imaging technique to probe unstained collagen-rich tissues like cornea. Moreover, corneal remodeling occurs in many diseases and precise characterization requires overcoming the limitations of conventional techniques. In this work, we focus on diabetes, which affects hundreds of million people worldwide and most often leads to diabetic retinopathy, with no early diagnostic tool. This study then aims to establish the potential of SHG microscopy for in situ detection and characterization of hyperglycemia-induced abnormalities in the Descemet's membrane, in the posterior cornea.<h4>Methodology/principal findings</h4>We studied corneas from age-matched control and Goto-Kakizaki rats, a spontaneous model of type 2 diabetes, and corneas from human donors with type 2 diabetes and without any diabetes. SHG imaging was compared to confocal microscopy, to histology characterization using conventional staining and transmitted light microscopy and to transmission electron microscopy. SHG imaging revealed collagen deposits in the Descemet's membrane of unstained corneas in a unique way compared to these gold standard techniques in ophthalmology. It provided background-free images of the three-dimensional interwoven distribution of the collagen deposits, with improved contrast compared to confocal microscopy. It also provided structural capability in intact corneas because of its high specificity to fibrillar collagen, with substantially larger field of view than transmission electron microscopy. Moreover, in vivo SHG imaging was demonstrated in Goto-Kakizaki rats.<h4>Conclusions/significance</h4>Our study shows unambiguously the high potential of SHG microscopy for three-dimensional characterization of structural abnormalities in unstained corneas. Furthermore, our demonstration of in vivo SHG imaging opens the way to long-term dynamical studies. This method should be easily generalized to other structural remodeling of the cornea and SHG microscopy should prove to be invaluable for in vivo corneal pathological studies.Gaël LatourLaura KowalczukMichèle SavoldelliJean-Louis BourgesKarsten PlamannFrancine Behar-CohenMarie-Claire Schanne-KleinPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 11, p e48388 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Gaël Latour Laura Kowalczuk Michèle Savoldelli Jean-Louis Bourges Karsten Plamann Francine Behar-Cohen Marie-Claire Schanne-Klein Hyperglycemia-induced abnormalities in rat and human corneas: the potential of second harmonic generation microscopy.
description	<h4>Background</h4>Second Harmonic Generation (SHG) microscopy recently appeared as an efficient optical imaging technique to probe unstained collagen-rich tissues like cornea. Moreover, corneal remodeling occurs in many diseases and precise characterization requires overcoming the limitations of conventional techniques. In this work, we focus on diabetes, which affects hundreds of million people worldwide and most often leads to diabetic retinopathy, with no early diagnostic tool. This study then aims to establish the potential of SHG microscopy for in situ detection and characterization of hyperglycemia-induced abnormalities in the Descemet's membrane, in the posterior cornea.<h4>Methodology/principal findings</h4>We studied corneas from age-matched control and Goto-Kakizaki rats, a spontaneous model of type 2 diabetes, and corneas from human donors with type 2 diabetes and without any diabetes. SHG imaging was compared to confocal microscopy, to histology characterization using conventional staining and transmitted light microscopy and to transmission electron microscopy. SHG imaging revealed collagen deposits in the Descemet's membrane of unstained corneas in a unique way compared to these gold standard techniques in ophthalmology. It provided background-free images of the three-dimensional interwoven distribution of the collagen deposits, with improved contrast compared to confocal microscopy. It also provided structural capability in intact corneas because of its high specificity to fibrillar collagen, with substantially larger field of view than transmission electron microscopy. Moreover, in vivo SHG imaging was demonstrated in Goto-Kakizaki rats.<h4>Conclusions/significance</h4>Our study shows unambiguously the high potential of SHG microscopy for three-dimensional characterization of structural abnormalities in unstained corneas. Furthermore, our demonstration of in vivo SHG imaging opens the way to long-term dynamical studies. This method should be easily generalized to other structural remodeling of the cornea and SHG microscopy should prove to be invaluable for in vivo corneal pathological studies.
format	article
author	Gaël Latour Laura Kowalczuk Michèle Savoldelli Jean-Louis Bourges Karsten Plamann Francine Behar-Cohen Marie-Claire Schanne-Klein
author_facet	Gaël Latour Laura Kowalczuk Michèle Savoldelli Jean-Louis Bourges Karsten Plamann Francine Behar-Cohen Marie-Claire Schanne-Klein
author_sort	Gaël Latour
title	Hyperglycemia-induced abnormalities in rat and human corneas: the potential of second harmonic generation microscopy.
title_short	Hyperglycemia-induced abnormalities in rat and human corneas: the potential of second harmonic generation microscopy.
title_full	Hyperglycemia-induced abnormalities in rat and human corneas: the potential of second harmonic generation microscopy.
title_fullStr	Hyperglycemia-induced abnormalities in rat and human corneas: the potential of second harmonic generation microscopy.
title_full_unstemmed	Hyperglycemia-induced abnormalities in rat and human corneas: the potential of second harmonic generation microscopy.
title_sort	hyperglycemia-induced abnormalities in rat and human corneas: the potential of second harmonic generation microscopy.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/e26093bb65464e42a9b464c1ea46c1bf
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Hyperglycemia-induced abnormalities in rat and human corneas: the potential of second harmonic generation microscopy.

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