A protocol for extracting immunolabeled murine cardiomyocytes of high-quality RNA by laser capture microdissection

We developed a highly efficient, ultrashort immunohistochemistry-laser capture microdissection (IHC-LMD) protocol, which allows microdissection of up to 250 single cardiomyocytes. Before LMD, murine hearts are excised, snap-frozen, and cryosectioned. RNA isolated from LMD material is of high RNA qua...

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Autores principales: Shaalan, Abeer K., Ellison-Hughes, Georgina M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8914385/
https://www.ncbi.nlm.nih.gov/pubmed/35284837
http://dx.doi.org/10.1016/j.xpro.2022.101231
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author Shaalan, Abeer K.
Ellison-Hughes, Georgina M.
author_facet Shaalan, Abeer K.
Ellison-Hughes, Georgina M.
author_sort Shaalan, Abeer K.
collection PubMed
description We developed a highly efficient, ultrashort immunohistochemistry-laser capture microdissection (IHC-LMD) protocol, which allows microdissection of up to 250 single cardiomyocytes. Before LMD, murine hearts are excised, snap-frozen, and cryosectioned. RNA isolated from LMD material is of high RNA quality, making it usable for gene expression analysis and RNA sequencing. Challenges and limitations of this protocol include visualization of the immunostaining and nuclei DAPI dye on the PEN slides, and timing and speed to limit RNA degradation as much as possible.
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spelling pubmed-89143852022-03-12 A protocol for extracting immunolabeled murine cardiomyocytes of high-quality RNA by laser capture microdissection Shaalan, Abeer K. Ellison-Hughes, Georgina M. STAR Protoc Protocol We developed a highly efficient, ultrashort immunohistochemistry-laser capture microdissection (IHC-LMD) protocol, which allows microdissection of up to 250 single cardiomyocytes. Before LMD, murine hearts are excised, snap-frozen, and cryosectioned. RNA isolated from LMD material is of high RNA quality, making it usable for gene expression analysis and RNA sequencing. Challenges and limitations of this protocol include visualization of the immunostaining and nuclei DAPI dye on the PEN slides, and timing and speed to limit RNA degradation as much as possible. Elsevier 2022-03-08 /pmc/articles/PMC8914385/ /pubmed/35284837 http://dx.doi.org/10.1016/j.xpro.2022.101231 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
Shaalan, Abeer K.
Ellison-Hughes, Georgina M.
A protocol for extracting immunolabeled murine cardiomyocytes of high-quality RNA by laser capture microdissection
title A protocol for extracting immunolabeled murine cardiomyocytes of high-quality RNA by laser capture microdissection
title_full A protocol for extracting immunolabeled murine cardiomyocytes of high-quality RNA by laser capture microdissection
title_fullStr A protocol for extracting immunolabeled murine cardiomyocytes of high-quality RNA by laser capture microdissection
title_full_unstemmed A protocol for extracting immunolabeled murine cardiomyocytes of high-quality RNA by laser capture microdissection
title_short A protocol for extracting immunolabeled murine cardiomyocytes of high-quality RNA by laser capture microdissection
title_sort protocol for extracting immunolabeled murine cardiomyocytes of high-quality rna by laser capture microdissection
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8914385/
https://www.ncbi.nlm.nih.gov/pubmed/35284837
http://dx.doi.org/10.1016/j.xpro.2022.101231
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