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Protocol for high-throughput electron tomography exemplified on centrioles in primary human plasma cells

Electron microscopy is the gold standard to characterize cellular ultrastructure. However, production of significant morphometrical data is highly limited by acquisition time. Here, we describe a semi-automated high-throughput strategy using single-axis serial section electron tomography to investig...

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Autores principales: Köhrer, Sebastian, Dittrich, Tobias, Schorb, Martin, Haberbosch, Isabella, Schwab, Yannick, Krämer, Alwin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10320260/
https://www.ncbi.nlm.nih.gov/pubmed/37354457
http://dx.doi.org/10.1016/j.xpro.2023.102373
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author Köhrer, Sebastian
Dittrich, Tobias
Schorb, Martin
Haberbosch, Isabella
Schwab, Yannick
Krämer, Alwin
author_facet Köhrer, Sebastian
Dittrich, Tobias
Schorb, Martin
Haberbosch, Isabella
Schwab, Yannick
Krämer, Alwin
author_sort Köhrer, Sebastian
collection PubMed
description Electron microscopy is the gold standard to characterize cellular ultrastructure. However, production of significant morphometrical data is highly limited by acquisition time. Here, we describe a semi-automated high-throughput strategy using single-axis serial section electron tomography to investigate and analyze centriole ultrastructure in bone-marrow-derived, primary human CD138(pos) plasma cells. The protocol comprises steps for electron microscopy sample preparation, semi-automated transmission electron microscopy screening, and screening evaluation for cells of interest. Thereafter, we detail tomography acquisition, data reconstruction, and joining. For complete details on the use and execution of this protocol, please refer to Dittrich et al.(1)
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spelling pubmed-103202602023-07-06 Protocol for high-throughput electron tomography exemplified on centrioles in primary human plasma cells Köhrer, Sebastian Dittrich, Tobias Schorb, Martin Haberbosch, Isabella Schwab, Yannick Krämer, Alwin STAR Protoc Protocol Electron microscopy is the gold standard to characterize cellular ultrastructure. However, production of significant morphometrical data is highly limited by acquisition time. Here, we describe a semi-automated high-throughput strategy using single-axis serial section electron tomography to investigate and analyze centriole ultrastructure in bone-marrow-derived, primary human CD138(pos) plasma cells. The protocol comprises steps for electron microscopy sample preparation, semi-automated transmission electron microscopy screening, and screening evaluation for cells of interest. Thereafter, we detail tomography acquisition, data reconstruction, and joining. For complete details on the use and execution of this protocol, please refer to Dittrich et al.(1) Elsevier 2023-06-22 /pmc/articles/PMC10320260/ /pubmed/37354457 http://dx.doi.org/10.1016/j.xpro.2023.102373 Text en © 2023 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
Köhrer, Sebastian
Dittrich, Tobias
Schorb, Martin
Haberbosch, Isabella
Schwab, Yannick
Krämer, Alwin
Protocol for high-throughput electron tomography exemplified on centrioles in primary human plasma cells
title Protocol for high-throughput electron tomography exemplified on centrioles in primary human plasma cells
title_full Protocol for high-throughput electron tomography exemplified on centrioles in primary human plasma cells
title_fullStr Protocol for high-throughput electron tomography exemplified on centrioles in primary human plasma cells
title_full_unstemmed Protocol for high-throughput electron tomography exemplified on centrioles in primary human plasma cells
title_short Protocol for high-throughput electron tomography exemplified on centrioles in primary human plasma cells
title_sort protocol for high-throughput electron tomography exemplified on centrioles in primary human plasma cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10320260/
https://www.ncbi.nlm.nih.gov/pubmed/37354457
http://dx.doi.org/10.1016/j.xpro.2023.102373
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