Analysis of target mRNAs in the fixed-frozen human brain using a modified BaseScope-ISH Assay protocol

We describe a modified BaseScope™ Assay protocol (ACDBio) for RNA in situ hybridization on fixed-frozen human brain tissue. The original protocol caused tissue detachment due to harsh tissue pre-treatment. We therefore optimized it to improve tissue stability while providing high stain quality in fr...

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Detalles Bibliográficos
Autores principales: Carisì, M. Carla, Howell, Owain W., Morgan, Alwena H., Davies, Jeffrey S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9720338/
https://www.ncbi.nlm.nih.gov/pubmed/36595945
http://dx.doi.org/10.1016/j.xpro.2022.101896
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author Carisì, M. Carla
Howell, Owain W.
Morgan, Alwena H.
Davies, Jeffrey S.
author_facet Carisì, M. Carla
Howell, Owain W.
Morgan, Alwena H.
Davies, Jeffrey S.
author_sort Carisì, M. Carla
collection PubMed
description We describe a modified BaseScope™ Assay protocol (ACDBio) for RNA in situ hybridization on fixed-frozen human brain tissue. The original protocol caused tissue detachment due to harsh tissue pre-treatment. We therefore optimized it to improve tissue stability while providing high stain quality in fragile post-mortem tissue from aged donors with advanced neurodegeneration. The main changes include two additional fixation steps and modifications to the pre-treatment protocol. We also describe tissue imaging and stain quantification using the open-source QuPath software. For complete details on the use and execution of this protocol, please refer to Hornsby et al. (2020).
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spelling pubmed-97203382022-12-06 Analysis of target mRNAs in the fixed-frozen human brain using a modified BaseScope-ISH Assay protocol Carisì, M. Carla Howell, Owain W. Morgan, Alwena H. Davies, Jeffrey S. STAR Protoc Protocol We describe a modified BaseScope™ Assay protocol (ACDBio) for RNA in situ hybridization on fixed-frozen human brain tissue. The original protocol caused tissue detachment due to harsh tissue pre-treatment. We therefore optimized it to improve tissue stability while providing high stain quality in fragile post-mortem tissue from aged donors with advanced neurodegeneration. The main changes include two additional fixation steps and modifications to the pre-treatment protocol. We also describe tissue imaging and stain quantification using the open-source QuPath software. For complete details on the use and execution of this protocol, please refer to Hornsby et al. (2020). Elsevier 2022-12-02 /pmc/articles/PMC9720338/ /pubmed/36595945 http://dx.doi.org/10.1016/j.xpro.2022.101896 Text en © 2022 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
Carisì, M. Carla
Howell, Owain W.
Morgan, Alwena H.
Davies, Jeffrey S.
Analysis of target mRNAs in the fixed-frozen human brain using a modified BaseScope-ISH Assay protocol
title Analysis of target mRNAs in the fixed-frozen human brain using a modified BaseScope-ISH Assay protocol
title_full Analysis of target mRNAs in the fixed-frozen human brain using a modified BaseScope-ISH Assay protocol
title_fullStr Analysis of target mRNAs in the fixed-frozen human brain using a modified BaseScope-ISH Assay protocol
title_full_unstemmed Analysis of target mRNAs in the fixed-frozen human brain using a modified BaseScope-ISH Assay protocol
title_short Analysis of target mRNAs in the fixed-frozen human brain using a modified BaseScope-ISH Assay protocol
title_sort analysis of target mrnas in the fixed-frozen human brain using a modified basescope-ish assay protocol
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9720338/
https://www.ncbi.nlm.nih.gov/pubmed/36595945
http://dx.doi.org/10.1016/j.xpro.2022.101896
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