TEM, SEM, and STEM-based immuno-CLEM workflows offer complementary advantages

Abstract Identifying endogenous tissue stem cells remains a key challenge in developmental and regenerative biology. To distinguish and molecularly characterise stem cell populations in large heterogeneous tissues, the combination of cytochemical cell markers with ultrastructural morphology is highl...

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Autores principales:	Viola Oorschot, Benjamin W. Lindsey, Jan Kaslin, Georg Ramm
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8ddcd5a7e12c41499aee1a0cc450ec98
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spelling	oai:doaj.org-article:8ddcd5a7e12c41499aee1a0cc450ec982021-12-02T15:23:10ZTEM, SEM, and STEM-based immuno-CLEM workflows offer complementary advantages10.1038/s41598-020-79637-92045-2322https://doaj.org/article/8ddcd5a7e12c41499aee1a0cc450ec982021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-79637-9https://doaj.org/toc/2045-2322Abstract Identifying endogenous tissue stem cells remains a key challenge in developmental and regenerative biology. To distinguish and molecularly characterise stem cell populations in large heterogeneous tissues, the combination of cytochemical cell markers with ultrastructural morphology is highly beneficial. Here, we realise this through workflows of multi-resolution immuno-correlative light and electron microscopy (iCLEM) methodologies. Taking advantage of the antigenicity preservation of the Tokuyasu technique, we have established robust protocols and workflows and provide a side-by-side comparison of iCLEM used in combination with scanning EM (SEM), scanning TEM (STEM), or transmission EM (TEM). Evaluation of the applications and advantages of each method highlights their practicality for the identification, quantification, and characterization of heterogeneous cell populations in small organisms, organs, or tissues in healthy and diseased states. The iCLEM techniques are broadly applicable and can use either genetically encoded or cytochemical markers on plant, animal and human tissues. We demonstrate how these protocols are particularly suited for investigating neural stem and progenitor cell populations of the vertebrate nervous system.Viola OorschotBenjamin W. LindseyJan KaslinGeorg RammNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-16 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Viola Oorschot Benjamin W. Lindsey Jan Kaslin Georg Ramm TEM, SEM, and STEM-based immuno-CLEM workflows offer complementary advantages
description	Abstract Identifying endogenous tissue stem cells remains a key challenge in developmental and regenerative biology. To distinguish and molecularly characterise stem cell populations in large heterogeneous tissues, the combination of cytochemical cell markers with ultrastructural morphology is highly beneficial. Here, we realise this through workflows of multi-resolution immuno-correlative light and electron microscopy (iCLEM) methodologies. Taking advantage of the antigenicity preservation of the Tokuyasu technique, we have established robust protocols and workflows and provide a side-by-side comparison of iCLEM used in combination with scanning EM (SEM), scanning TEM (STEM), or transmission EM (TEM). Evaluation of the applications and advantages of each method highlights their practicality for the identification, quantification, and characterization of heterogeneous cell populations in small organisms, organs, or tissues in healthy and diseased states. The iCLEM techniques are broadly applicable and can use either genetically encoded or cytochemical markers on plant, animal and human tissues. We demonstrate how these protocols are particularly suited for investigating neural stem and progenitor cell populations of the vertebrate nervous system.
format	article
author	Viola Oorschot Benjamin W. Lindsey Jan Kaslin Georg Ramm
author_facet	Viola Oorschot Benjamin W. Lindsey Jan Kaslin Georg Ramm
author_sort	Viola Oorschot
title	TEM, SEM, and STEM-based immuno-CLEM workflows offer complementary advantages
title_short	TEM, SEM, and STEM-based immuno-CLEM workflows offer complementary advantages
title_full	TEM, SEM, and STEM-based immuno-CLEM workflows offer complementary advantages
title_fullStr	TEM, SEM, and STEM-based immuno-CLEM workflows offer complementary advantages
title_full_unstemmed	TEM, SEM, and STEM-based immuno-CLEM workflows offer complementary advantages
title_sort	tem, sem, and stem-based immuno-clem workflows offer complementary advantages
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/8ddcd5a7e12c41499aee1a0cc450ec98
work_keys_str_mv	AT violaoorschot temsemandstembasedimmunoclemworkflowsoffercomplementaryadvantages AT benjaminwlindsey temsemandstembasedimmunoclemworkflowsoffercomplementaryadvantages AT jankaslin temsemandstembasedimmunoclemworkflowsoffercomplementaryadvantages AT georgramm temsemandstembasedimmunoclemworkflowsoffercomplementaryadvantages
_version_	1718387277384646656

TEM, SEM, and STEM-based immuno-CLEM workflows offer complementary advantages

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