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Modified CLARITY Achieving Faster and Better Intact Mouse Brain Clearing and Immunostaining
CLARITY is a hydrogel embedding clearing method that has the advantages of transparency, different tissue compatibility and immunostaining compatibility. However, there are also some limitations to CLARITY as it requires a long time to achieve transparency, and the electrophoresis clearing is comple...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6646319/ https://www.ncbi.nlm.nih.gov/pubmed/31332235 http://dx.doi.org/10.1038/s41598-019-46814-4 |
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author | Du, Hao Hou, Peihong Wang, Liting Wang, Zhongke Li, Qiyu |
author_facet | Du, Hao Hou, Peihong Wang, Liting Wang, Zhongke Li, Qiyu |
author_sort | Du, Hao |
collection | PubMed |
description | CLARITY is a hydrogel embedding clearing method that has the advantages of transparency, different tissue compatibility and immunostaining compatibility. However, there are also some limitations to CLARITY as it requires a long time to achieve transparency, and the electrophoresis clearing is complex. Therefore, we aimed to simplify the electrophoresis system and shorten the processing time of CLARITY. In our study, we developed a non-circulation electrophoresis system to achieve easier manipulation of electrophoresis clearing. We modified the original CLARITY protocol in hydrogel embedding methods, clearing buffer and immunostaining. When comparing brains processed by our modified method or the original protocol, we found our modifications permit faster and more efficient clearing and labeling. Moreover, we developed a new clearing method named Passive pRe-Electrophroresis CLARITY (PRE-CLARITY) and a new immunostaining method named Centrifugation-Expansion staining (CEx staining). PRE-CLARITY achieved faster clearing and higher transparency, and CEx staining accomplished intact mouse brain labeling faster. With our modifications to CLARITY, we accomplished intact mouse brain clearing and immunostaining within one week, while this requires weeks to months with the original CLARITY. Our studies would allow high-content tracing and analysis of intact brain or other large-scale samples in a short time. |
format | Online Article Text |
id | pubmed-6646319 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-66463192019-07-29 Modified CLARITY Achieving Faster and Better Intact Mouse Brain Clearing and Immunostaining Du, Hao Hou, Peihong Wang, Liting Wang, Zhongke Li, Qiyu Sci Rep Article CLARITY is a hydrogel embedding clearing method that has the advantages of transparency, different tissue compatibility and immunostaining compatibility. However, there are also some limitations to CLARITY as it requires a long time to achieve transparency, and the electrophoresis clearing is complex. Therefore, we aimed to simplify the electrophoresis system and shorten the processing time of CLARITY. In our study, we developed a non-circulation electrophoresis system to achieve easier manipulation of electrophoresis clearing. We modified the original CLARITY protocol in hydrogel embedding methods, clearing buffer and immunostaining. When comparing brains processed by our modified method or the original protocol, we found our modifications permit faster and more efficient clearing and labeling. Moreover, we developed a new clearing method named Passive pRe-Electrophroresis CLARITY (PRE-CLARITY) and a new immunostaining method named Centrifugation-Expansion staining (CEx staining). PRE-CLARITY achieved faster clearing and higher transparency, and CEx staining accomplished intact mouse brain labeling faster. With our modifications to CLARITY, we accomplished intact mouse brain clearing and immunostaining within one week, while this requires weeks to months with the original CLARITY. Our studies would allow high-content tracing and analysis of intact brain or other large-scale samples in a short time. Nature Publishing Group UK 2019-07-22 /pmc/articles/PMC6646319/ /pubmed/31332235 http://dx.doi.org/10.1038/s41598-019-46814-4 Text en © The Author(s) 2019 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/. |
spellingShingle | Article Du, Hao Hou, Peihong Wang, Liting Wang, Zhongke Li, Qiyu Modified CLARITY Achieving Faster and Better Intact Mouse Brain Clearing and Immunostaining |
title | Modified CLARITY Achieving Faster and Better Intact Mouse Brain Clearing and Immunostaining |
title_full | Modified CLARITY Achieving Faster and Better Intact Mouse Brain Clearing and Immunostaining |
title_fullStr | Modified CLARITY Achieving Faster and Better Intact Mouse Brain Clearing and Immunostaining |
title_full_unstemmed | Modified CLARITY Achieving Faster and Better Intact Mouse Brain Clearing and Immunostaining |
title_short | Modified CLARITY Achieving Faster and Better Intact Mouse Brain Clearing and Immunostaining |
title_sort | modified clarity achieving faster and better intact mouse brain clearing and immunostaining |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6646319/ https://www.ncbi.nlm.nih.gov/pubmed/31332235 http://dx.doi.org/10.1038/s41598-019-46814-4 |
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