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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Mary Ann Liebert, Inc.
2016
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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. |
format | Online Article Text |
id | pubmed-4922008 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Mary Ann Liebert, Inc. |
record_format | MEDLINE/PubMed |
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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