Approaches to Using the Chameleon: Robust, Automated, Fast-Plunge cryoEM Specimen Preparation
<jats:p>The specimen preparation process is a key determinant in the success of any cryo electron microscopy (cryoEM) structural study and until recently had remained largely unchanged from the initial designs of Jacques Dubochet and others in the 1980s. The process has transformed structural...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media SA
2022
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| Online Access: | https://hdl.handle.net/1721.1/146787 |
| _version_ | 1826212992531824640 |
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| author | Levitz, Talya S Weckener, Miriam Fong, Ivan Naismith, James H Drennan, Catherine L Brignole, Edward J Clare, Daniel K Darrow, Michele C |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Levitz, Talya S Weckener, Miriam Fong, Ivan Naismith, James H Drennan, Catherine L Brignole, Edward J Clare, Daniel K Darrow, Michele C |
| author_sort | Levitz, Talya S |
| collection | MIT |
| description | <jats:p>The specimen preparation process is a key determinant in the success of any cryo electron microscopy (cryoEM) structural study and until recently had remained largely unchanged from the initial designs of Jacques Dubochet and others in the 1980s. The process has transformed structural biology, but it is largely manual and can require extensive optimisation for each protein sample. The chameleon instrument with its self-wicking grids and fast-plunge freezing represents a shift towards a robust, automated, and highly controllable future for specimen preparation. However, these new technologies require new workflows and an understanding of their limitations and strengths. As early adopters of the chameleon technology, we report on our experiences and lessons learned through case studies. We use these to make recommendations for the benefit of future users of the chameleon system and the field of cryoEM specimen preparation generally.</jats:p> |
| first_indexed | 2024-09-23T15:41:56Z |
| format | Article |
| id | mit-1721.1/146787 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T15:41:56Z |
| publishDate | 2022 |
| publisher | Frontiers Media SA |
| record_format | dspace |
| spelling | mit-1721.1/1467872022-12-08T03:05:33Z Approaches to Using the Chameleon: Robust, Automated, Fast-Plunge cryoEM Specimen Preparation Levitz, Talya S Weckener, Miriam Fong, Ivan Naismith, James H Drennan, Catherine L Brignole, Edward J Clare, Daniel K Darrow, Michele C Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry <jats:p>The specimen preparation process is a key determinant in the success of any cryo electron microscopy (cryoEM) structural study and until recently had remained largely unchanged from the initial designs of Jacques Dubochet and others in the 1980s. The process has transformed structural biology, but it is largely manual and can require extensive optimisation for each protein sample. The chameleon instrument with its self-wicking grids and fast-plunge freezing represents a shift towards a robust, automated, and highly controllable future for specimen preparation. However, these new technologies require new workflows and an understanding of their limitations and strengths. As early adopters of the chameleon technology, we report on our experiences and lessons learned through case studies. We use these to make recommendations for the benefit of future users of the chameleon system and the field of cryoEM specimen preparation generally.</jats:p> 2022-12-07T18:18:38Z 2022-12-07T18:18:38Z 2022 2022-12-07T18:08:22Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/146787 Levitz, Talya S, Weckener, Miriam, Fong, Ivan, Naismith, James H, Drennan, Catherine L et al. 2022. "Approaches to Using the Chameleon: Robust, Automated, Fast-Plunge cryoEM Specimen Preparation." Frontiers in Molecular Biosciences, 9. en 10.3389/FMOLB.2022.903148 Frontiers in Molecular Biosciences Creative Commons Attribution 4.0 International license https://creativecommons.org/licenses/by/4.0/ application/pdf Frontiers Media SA Frontiers |
| spellingShingle | Levitz, Talya S Weckener, Miriam Fong, Ivan Naismith, James H Drennan, Catherine L Brignole, Edward J Clare, Daniel K Darrow, Michele C Approaches to Using the Chameleon: Robust, Automated, Fast-Plunge cryoEM Specimen Preparation |
| title | Approaches to Using the Chameleon: Robust, Automated, Fast-Plunge cryoEM Specimen Preparation |
| title_full | Approaches to Using the Chameleon: Robust, Automated, Fast-Plunge cryoEM Specimen Preparation |
| title_fullStr | Approaches to Using the Chameleon: Robust, Automated, Fast-Plunge cryoEM Specimen Preparation |
| title_full_unstemmed | Approaches to Using the Chameleon: Robust, Automated, Fast-Plunge cryoEM Specimen Preparation |
| title_short | Approaches to Using the Chameleon: Robust, Automated, Fast-Plunge cryoEM Specimen Preparation |
| title_sort | approaches to using the chameleon robust automated fast plunge cryoem specimen preparation |
| url | https://hdl.handle.net/1721.1/146787 |
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