Academic Journal
Direct deposition on pre-cooled grids: a new approach to sub-millisecond cryoEM grid preparation.
| Τίτλος: | Direct deposition on pre-cooled grids: a new approach to sub-millisecond cryoEM grid preparation. |
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| Συγγραφείς: | Klebl DP; School of Biomedical Sciences, Faculty of Biomedical Sciences and Astbury Centre for Structural and Molecular Biology, University of Leeds, Leeds, United Kingdom., Kay RW; School of Mechanical Engineering, University of Leeds, Leeds, United Kingdom., McMillan S; School of Biomedical Sciences, Faculty of Biomedical Sciences and Astbury Centre for Structural and Molecular Biology, University of Leeds, Leeds, United Kingdom., Sobott F; School of Molecular and Cellular Biology, Faculty of Biological Sciences and Astbury Centre for Structural and Molecular Biology, University of Leeds, Leeds, United Kingdom., Kapur N; School of Mechanical Engineering, University of Leeds, Leeds, United Kingdom., Muench SP; School of Biomedical Sciences, Faculty of Biomedical Sciences and Astbury Centre for Structural and Molecular Biology, University of Leeds, Leeds, United Kingdom. |
| Πηγή: | Acta crystallographica. Section D, Structural biology [Acta Crystallogr D Struct Biol] 2026 Sep 01; Vol. 82 (Pt 9), pp. 1056-1061. Date of Electronic Publication: 2026 Aug 11. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: John Wiley & Sons Country of Publication: United States NLM ID: 101676043 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2059-7983 (Electronic) Linking ISSN: 20597983 NLM ISO Abbreviation: Acta Crystallogr D Struct Biol Subsets: MEDLINE |
| Imprint Name(s): | Publication: <2018-> : Medford, MA : John Wiley & Sons Original Publication: [Malden, MA] : John Wiley & Sons, [2016]- |
| Ιατρικοί όροι (MeSH): | Cryoelectron Microscopy*/methods , Cryoelectron Microscopy*/instrumentation , Specimen Handling*/methods |
| Περίληψη: | There has been a rapid expansion in new approaches to sample preparation for single-particle cryoEM. One drive has been around speeding up the process both for the application of time-resolved studies and in outrunning some of the negative effects which may come from the sample interacting with the air-water interface, such as degradation and preferred orientation. Here, we set out to significantly improve the speed of grid preparation by removing the plunging stage and directly spraying onto pre-cooled grids which are situated in the cryogen, allowing us to produce grids where two samples are mixed and frozen on the grid in less than 1 ms. We have previously shown a preliminary proof of principle with this new approach and here we provide a detailed description of the approach taken and the methodology behind it. This approach allows us to move beyond the millisecond timescale to open up more systems to time-resolved EM and potentially improve samples. (open access.) |
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| Grant Information: | United Kingdom WT_ Wellcome Trust; 198466/Z/15/Z United Kingdom WT_ Wellcome Trust; UKR11958 United Kingdom BB_ Biotechnology and Biological Sciences Research Council |
| Contributed Indexing: | Keywords: cryoEM; structural biology; time-resolved cryoEM |
| Entry Date(s): | Date Created: 20260811 Date Completed: 20260901 Latest Revision: 20260903 |
| Update Code: | 20260903 |
| PubMed Central ID: | PMC13532694 |
| DOI: | 10.1107/S2059798326007126 |
| PMID: | 42579337 |
| Βάση Δεδομένων: | MEDLINE |
| ISSN: | 2059-7983 |
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| DOI: | 10.1107/S2059798326007126 |