Step-by-step preparation of mouse eye sections for routine histology, immunofluorescence, and RNA in situ hybridization multiplexing

It can be challenging to maintain tissue integrity using established histology protocols. Here, we describe a protocol composed of Hartman’s fixation, window technique, microwave-based tissue processing, optimized depigmentation, and antigen retrieval pretreatment. This is followed by the ViewRNA si...

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Detalles Bibliográficos
Autores principales: Pang, Ji, Thomas, Nancy, Tsuchiya, Dai, Parmely, Tari, Yan, Deyue, Xie, Ting, Wang, Yongfu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8585658/
https://www.ncbi.nlm.nih.gov/pubmed/34806041
http://dx.doi.org/10.1016/j.xpro.2021.100879
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author Pang, Ji
Thomas, Nancy
Tsuchiya, Dai
Parmely, Tari
Yan, Deyue
Xie, Ting
Wang, Yongfu
author_facet Pang, Ji
Thomas, Nancy
Tsuchiya, Dai
Parmely, Tari
Yan, Deyue
Xie, Ting
Wang, Yongfu
author_sort Pang, Ji
collection PubMed
description It can be challenging to maintain tissue integrity using established histology protocols. Here, we describe a protocol composed of Hartman’s fixation, window technique, microwave-based tissue processing, optimized depigmentation, and antigen retrieval pretreatment. This is followed by the ViewRNA single-molecule fluorescence in situ hybridization and immunofluorescence techniques to optimize routine histological staining and molecular histology multiplexing assays. Our protocol is highly reproducible in any laboratory and may decrease animal usage and lab resource expenditure. For complete details on the use and execution of this protocol, please refer to Pang et al. (2021).
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spelling pubmed-85856582021-11-18 Step-by-step preparation of mouse eye sections for routine histology, immunofluorescence, and RNA in situ hybridization multiplexing Pang, Ji Thomas, Nancy Tsuchiya, Dai Parmely, Tari Yan, Deyue Xie, Ting Wang, Yongfu STAR Protoc Protocol It can be challenging to maintain tissue integrity using established histology protocols. Here, we describe a protocol composed of Hartman’s fixation, window technique, microwave-based tissue processing, optimized depigmentation, and antigen retrieval pretreatment. This is followed by the ViewRNA single-molecule fluorescence in situ hybridization and immunofluorescence techniques to optimize routine histological staining and molecular histology multiplexing assays. Our protocol is highly reproducible in any laboratory and may decrease animal usage and lab resource expenditure. For complete details on the use and execution of this protocol, please refer to Pang et al. (2021). Elsevier 2021-11-05 /pmc/articles/PMC8585658/ /pubmed/34806041 http://dx.doi.org/10.1016/j.xpro.2021.100879 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Pang, Ji
Thomas, Nancy
Tsuchiya, Dai
Parmely, Tari
Yan, Deyue
Xie, Ting
Wang, Yongfu
Step-by-step preparation of mouse eye sections for routine histology, immunofluorescence, and RNA in situ hybridization multiplexing
title Step-by-step preparation of mouse eye sections for routine histology, immunofluorescence, and RNA in situ hybridization multiplexing
title_full Step-by-step preparation of mouse eye sections for routine histology, immunofluorescence, and RNA in situ hybridization multiplexing
title_fullStr Step-by-step preparation of mouse eye sections for routine histology, immunofluorescence, and RNA in situ hybridization multiplexing
title_full_unstemmed Step-by-step preparation of mouse eye sections for routine histology, immunofluorescence, and RNA in situ hybridization multiplexing
title_short Step-by-step preparation of mouse eye sections for routine histology, immunofluorescence, and RNA in situ hybridization multiplexing
title_sort step-by-step preparation of mouse eye sections for routine histology, immunofluorescence, and rna in situ hybridization multiplexing
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8585658/
https://www.ncbi.nlm.nih.gov/pubmed/34806041
http://dx.doi.org/10.1016/j.xpro.2021.100879
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