Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue

We describe an optimized smFISH protocol for the intact pancreas. The protocol is adapted from Lyubimova et al. (2013), a generic tissue smFISH protocol that works for most tissues but not the pancreas. The main changes implemented include increasing the period of mRNA denaturation from 5 min to at...

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Detalles Bibliográficos
Autores principales: Farack, Lydia, Itzkovitz, Shalev
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580104/
https://www.ncbi.nlm.nih.gov/pubmed/33111069
http://dx.doi.org/10.1016/j.xpro.2019.100007
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author Farack, Lydia
Itzkovitz, Shalev
author_facet Farack, Lydia
Itzkovitz, Shalev
author_sort Farack, Lydia
collection PubMed
description We describe an optimized smFISH protocol for the intact pancreas. The protocol is adapted from Lyubimova et al. (2013), a generic tissue smFISH protocol that works for most tissues but not the pancreas. The main changes implemented include increasing the period of mRNA denaturation from 5 min to at least 3 h and increasing formamide concentrations from 10% to 30%. These modifications yield sensitive single mRNA visualization that is comparable to those achieved in other tissues using the standard protocol. For complete details on the use and execution of this protocol, please refer to Farack et al., 2018, Farack et al., 2019.
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spelling pubmed-75801042020-10-26 Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue Farack, Lydia Itzkovitz, Shalev STAR Protoc Protocol We describe an optimized smFISH protocol for the intact pancreas. The protocol is adapted from Lyubimova et al. (2013), a generic tissue smFISH protocol that works for most tissues but not the pancreas. The main changes implemented include increasing the period of mRNA denaturation from 5 min to at least 3 h and increasing formamide concentrations from 10% to 30%. These modifications yield sensitive single mRNA visualization that is comparable to those achieved in other tissues using the standard protocol. For complete details on the use and execution of this protocol, please refer to Farack et al., 2018, Farack et al., 2019. Elsevier 2020-06-03 /pmc/articles/PMC7580104/ /pubmed/33111069 http://dx.doi.org/10.1016/j.xpro.2019.100007 Text en © 2020. http://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
Farack, Lydia
Itzkovitz, Shalev
Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue
title Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue
title_full Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue
title_fullStr Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue
title_full_unstemmed Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue
title_short Protocol for Single-Molecule Fluorescence In Situ Hybridization for Intact Pancreatic Tissue
title_sort protocol for single-molecule fluorescence in situ hybridization for intact pancreatic tissue
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580104/
https://www.ncbi.nlm.nih.gov/pubmed/33111069
http://dx.doi.org/10.1016/j.xpro.2019.100007
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