Cargando…
Approaches to altering particle distributions in cryo-electron microscopy sample preparation
Cryo-electron microscopy (cryo-EM) can now be used to determine high-resolution structural information on a diverse range of biological specimens. Recent advances have been driven primarily by developments in microscopes and detectors, and through advances in image-processing software. However, for...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6096488/ https://www.ncbi.nlm.nih.gov/pubmed/29872006 http://dx.doi.org/10.1107/S2059798318006496 |
_version_ | 1783348111020654592 |
---|---|
author | Drulyte, Ieva Johnson, Rachel M. Hesketh, Emma L. Hurdiss, Daniel L. Scarff, Charlotte A. Porav, Sebastian A. Ranson, Neil A. Muench, Stephen P. Thompson, Rebecca F. |
author_facet | Drulyte, Ieva Johnson, Rachel M. Hesketh, Emma L. Hurdiss, Daniel L. Scarff, Charlotte A. Porav, Sebastian A. Ranson, Neil A. Muench, Stephen P. Thompson, Rebecca F. |
author_sort | Drulyte, Ieva |
collection | PubMed |
description | Cryo-electron microscopy (cryo-EM) can now be used to determine high-resolution structural information on a diverse range of biological specimens. Recent advances have been driven primarily by developments in microscopes and detectors, and through advances in image-processing software. However, for many single-particle cryo-EM projects, major bottlenecks currently remain at the sample-preparation stage; obtaining cryo-EM grids of sufficient quality for high-resolution single-particle analysis can require the careful optimization of many variables. Common hurdles to overcome include problems associated with the sample itself (buffer components, labile complexes), sample distribution (obtaining the correct concentration, affinity for the support film), preferred orientation, and poor reproducibility of the grid-making process within and between batches. This review outlines a number of methodologies used within the electron-microscopy community to address these challenges, providing a range of approaches which may aid in obtaining optimal grids for high-resolution data collection. |
format | Online Article Text |
id | pubmed-6096488 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-60964882018-08-24 Approaches to altering particle distributions in cryo-electron microscopy sample preparation Drulyte, Ieva Johnson, Rachel M. Hesketh, Emma L. Hurdiss, Daniel L. Scarff, Charlotte A. Porav, Sebastian A. Ranson, Neil A. Muench, Stephen P. Thompson, Rebecca F. Acta Crystallogr D Struct Biol Research Papers Cryo-electron microscopy (cryo-EM) can now be used to determine high-resolution structural information on a diverse range of biological specimens. Recent advances have been driven primarily by developments in microscopes and detectors, and through advances in image-processing software. However, for many single-particle cryo-EM projects, major bottlenecks currently remain at the sample-preparation stage; obtaining cryo-EM grids of sufficient quality for high-resolution single-particle analysis can require the careful optimization of many variables. Common hurdles to overcome include problems associated with the sample itself (buffer components, labile complexes), sample distribution (obtaining the correct concentration, affinity for the support film), preferred orientation, and poor reproducibility of the grid-making process within and between batches. This review outlines a number of methodologies used within the electron-microscopy community to address these challenges, providing a range of approaches which may aid in obtaining optimal grids for high-resolution data collection. International Union of Crystallography 2018-05-18 /pmc/articles/PMC6096488/ /pubmed/29872006 http://dx.doi.org/10.1107/S2059798318006496 Text en © Drulyte et al. 2018 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.http://creativecommons.org/licenses/by/2.0/uk/ |
spellingShingle | Research Papers Drulyte, Ieva Johnson, Rachel M. Hesketh, Emma L. Hurdiss, Daniel L. Scarff, Charlotte A. Porav, Sebastian A. Ranson, Neil A. Muench, Stephen P. Thompson, Rebecca F. Approaches to altering particle distributions in cryo-electron microscopy sample preparation |
title | Approaches to altering particle distributions in cryo-electron microscopy sample preparation |
title_full | Approaches to altering particle distributions in cryo-electron microscopy sample preparation |
title_fullStr | Approaches to altering particle distributions in cryo-electron microscopy sample preparation |
title_full_unstemmed | Approaches to altering particle distributions in cryo-electron microscopy sample preparation |
title_short | Approaches to altering particle distributions in cryo-electron microscopy sample preparation |
title_sort | approaches to altering particle distributions in cryo-electron microscopy sample preparation |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6096488/ https://www.ncbi.nlm.nih.gov/pubmed/29872006 http://dx.doi.org/10.1107/S2059798318006496 |
work_keys_str_mv | AT drulyteieva approachestoalteringparticledistributionsincryoelectronmicroscopysamplepreparation AT johnsonrachelm approachestoalteringparticledistributionsincryoelectronmicroscopysamplepreparation AT heskethemmal approachestoalteringparticledistributionsincryoelectronmicroscopysamplepreparation AT hurdissdaniell approachestoalteringparticledistributionsincryoelectronmicroscopysamplepreparation AT scarffcharlottea approachestoalteringparticledistributionsincryoelectronmicroscopysamplepreparation AT poravsebastiana approachestoalteringparticledistributionsincryoelectronmicroscopysamplepreparation AT ransonneila approachestoalteringparticledistributionsincryoelectronmicroscopysamplepreparation AT muenchstephenp approachestoalteringparticledistributionsincryoelectronmicroscopysamplepreparation AT thompsonrebeccaf approachestoalteringparticledistributionsincryoelectronmicroscopysamplepreparation |