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Protocol for the use of signal amplification by exchange reaction-fluorescence in situ hybridization on adult formalin-fixed paraffin-embedded mouse lung tissue

Fluorescence in situ hybridization (FISH) is a useful tool for analyzing RNA expression, but difficulties arise with low-abundance RNA and in tissues that are formalin-fixed paraffin-embedded (FFPE) because reagents can be expensive. In this protocol, we adapt a previously designed FISH amplificatio...

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Autores principales: Warren, Rachel, Shaik, Aftab, Teubner, Lauren, Lyu, Handeng, De Langhe, Stijn
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10276273/
https://www.ncbi.nlm.nih.gov/pubmed/37302070
http://dx.doi.org/10.1016/j.xpro.2023.102353
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author Warren, Rachel
Shaik, Aftab
Teubner, Lauren
Lyu, Handeng
De Langhe, Stijn
author_facet Warren, Rachel
Shaik, Aftab
Teubner, Lauren
Lyu, Handeng
De Langhe, Stijn
author_sort Warren, Rachel
collection PubMed
description Fluorescence in situ hybridization (FISH) is a useful tool for analyzing RNA expression, but difficulties arise with low-abundance RNA and in tissues that are formalin-fixed paraffin-embedded (FFPE) because reagents can be expensive. In this protocol, we adapt a previously designed FISH amplification protocol (SABER [signal amplification by exchange reaction]) for adult mouse FFPE lung sections by using probes that are extended and branched to amplify the signal. We combine FISH and immunostaining to identify cell-specific RNA. For complete details on the use and execution of this protocol, please refer to Kishi et al.(1) and Lyu et al.(2)
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spelling pubmed-102762732023-06-18 Protocol for the use of signal amplification by exchange reaction-fluorescence in situ hybridization on adult formalin-fixed paraffin-embedded mouse lung tissue Warren, Rachel Shaik, Aftab Teubner, Lauren Lyu, Handeng De Langhe, Stijn STAR Protoc Protocol Fluorescence in situ hybridization (FISH) is a useful tool for analyzing RNA expression, but difficulties arise with low-abundance RNA and in tissues that are formalin-fixed paraffin-embedded (FFPE) because reagents can be expensive. In this protocol, we adapt a previously designed FISH amplification protocol (SABER [signal amplification by exchange reaction]) for adult mouse FFPE lung sections by using probes that are extended and branched to amplify the signal. We combine FISH and immunostaining to identify cell-specific RNA. For complete details on the use and execution of this protocol, please refer to Kishi et al.(1) and Lyu et al.(2) Elsevier 2023-06-09 /pmc/articles/PMC10276273/ /pubmed/37302070 http://dx.doi.org/10.1016/j.xpro.2023.102353 Text en © 2023. 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
Warren, Rachel
Shaik, Aftab
Teubner, Lauren
Lyu, Handeng
De Langhe, Stijn
Protocol for the use of signal amplification by exchange reaction-fluorescence in situ hybridization on adult formalin-fixed paraffin-embedded mouse lung tissue
title Protocol for the use of signal amplification by exchange reaction-fluorescence in situ hybridization on adult formalin-fixed paraffin-embedded mouse lung tissue
title_full Protocol for the use of signal amplification by exchange reaction-fluorescence in situ hybridization on adult formalin-fixed paraffin-embedded mouse lung tissue
title_fullStr Protocol for the use of signal amplification by exchange reaction-fluorescence in situ hybridization on adult formalin-fixed paraffin-embedded mouse lung tissue
title_full_unstemmed Protocol for the use of signal amplification by exchange reaction-fluorescence in situ hybridization on adult formalin-fixed paraffin-embedded mouse lung tissue
title_short Protocol for the use of signal amplification by exchange reaction-fluorescence in situ hybridization on adult formalin-fixed paraffin-embedded mouse lung tissue
title_sort protocol for the use of signal amplification by exchange reaction-fluorescence in situ hybridization on adult formalin-fixed paraffin-embedded mouse lung tissue
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10276273/
https://www.ncbi.nlm.nih.gov/pubmed/37302070
http://dx.doi.org/10.1016/j.xpro.2023.102353
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