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Enabling Multiphoton and Second Harmonic Generation Imaging in Paraffin-Embedded and Histologically Stained Sections

Nonlinear microscopy, namely multiphoton imaging and second harmonic generation (SHG), is an established noninvasive technique useful for the imaging of extracellular matrix (ECM). Typically, measurements are performed in vivo on freshly excised tissues or biopsies. In this article, we describe the...

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Autores principales: Monaghan, Michael G., Kroll, Sebastian, Brucker, Sara Y., Schenke-Layland, Katja
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Mary Ann Liebert, Inc. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4922008/
https://www.ncbi.nlm.nih.gov/pubmed/27018844
http://dx.doi.org/10.1089/ten.tec.2016.0071
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author Monaghan, Michael G.
Kroll, Sebastian
Brucker, Sara Y.
Schenke-Layland, Katja
author_facet Monaghan, Michael G.
Kroll, Sebastian
Brucker, Sara Y.
Schenke-Layland, Katja
author_sort Monaghan, Michael G.
collection PubMed
description Nonlinear microscopy, namely multiphoton imaging and second harmonic generation (SHG), is an established noninvasive technique useful for the imaging of extracellular matrix (ECM). Typically, measurements are performed in vivo on freshly excised tissues or biopsies. In this article, we describe the effect of rehydrating paraffin-embedded sections on multiphoton and SHG emission signals and the acquisition of nonlinear images from hematoxylin and eosin (H&E)-stained sections before and after a destaining protocol. Our results reveal that bringing tissue sections to a physiological state yields a significant improvement in nonlinear signals, particularly in SHG. Additionally, the destaining of sections previously processed with H&E staining significantly improves their SHG emission signals during imaging, thereby allowing sufficient analysis of collagen in these sections. These results are important for researchers and pathologists to obtain additional information from paraffin-embedded tissues and archived samples to perform retrospective analysis of the ECM or gain additional information from rare samples.
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spelling pubmed-49220082016-07-06 Enabling Multiphoton and Second Harmonic Generation Imaging in Paraffin-Embedded and Histologically Stained Sections Monaghan, Michael G. Kroll, Sebastian Brucker, Sara Y. Schenke-Layland, Katja Tissue Eng Part C Methods Article Nonlinear microscopy, namely multiphoton imaging and second harmonic generation (SHG), is an established noninvasive technique useful for the imaging of extracellular matrix (ECM). Typically, measurements are performed in vivo on freshly excised tissues or biopsies. In this article, we describe the effect of rehydrating paraffin-embedded sections on multiphoton and SHG emission signals and the acquisition of nonlinear images from hematoxylin and eosin (H&E)-stained sections before and after a destaining protocol. Our results reveal that bringing tissue sections to a physiological state yields a significant improvement in nonlinear signals, particularly in SHG. Additionally, the destaining of sections previously processed with H&E staining significantly improves their SHG emission signals during imaging, thereby allowing sufficient analysis of collagen in these sections. These results are important for researchers and pathologists to obtain additional information from paraffin-embedded tissues and archived samples to perform retrospective analysis of the ECM or gain additional information from rare samples. Mary Ann Liebert, Inc. 2016-06-01 2016-04-22 /pmc/articles/PMC4922008/ /pubmed/27018844 http://dx.doi.org/10.1089/ten.tec.2016.0071 Text en © Michael G. Monaghan, et al., 2016; Published by Mary Ann Liebert, Inc. This Open Access article is distributed under the terms of the Creative Commons Attribution Noncommercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.
spellingShingle Article
Monaghan, Michael G.
Kroll, Sebastian
Brucker, Sara Y.
Schenke-Layland, Katja
Enabling Multiphoton and Second Harmonic Generation Imaging in Paraffin-Embedded and Histologically Stained Sections
title Enabling Multiphoton and Second Harmonic Generation Imaging in Paraffin-Embedded and Histologically Stained Sections
title_full Enabling Multiphoton and Second Harmonic Generation Imaging in Paraffin-Embedded and Histologically Stained Sections
title_fullStr Enabling Multiphoton and Second Harmonic Generation Imaging in Paraffin-Embedded and Histologically Stained Sections
title_full_unstemmed Enabling Multiphoton and Second Harmonic Generation Imaging in Paraffin-Embedded and Histologically Stained Sections
title_short Enabling Multiphoton and Second Harmonic Generation Imaging in Paraffin-Embedded and Histologically Stained Sections
title_sort enabling multiphoton and second harmonic generation imaging in paraffin-embedded and histologically stained sections
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4922008/
https://www.ncbi.nlm.nih.gov/pubmed/27018844
http://dx.doi.org/10.1089/ten.tec.2016.0071
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