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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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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) |
format | Online Article Text |
id | pubmed-10320260 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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