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| 001 | 1032201 | ||
| 005 | 20250203133217.0 | ||
| 024 | 7 | _ | |a 10.1016/j.jsb.2024.108139 |2 doi |
| 024 | 7 | _ | |a 1047-8477 |2 ISSN |
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| 024 | 7 | _ | |a 10.34734/FZJ-2024-06070 |2 datacite_doi |
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| 100 | 1 | _ | |a Henderikx, Rene J. M. |0 0009-0005-1709-7587 |b 0 |e Corresponding author |
| 245 | _ | _ | |a Ice thickness control and measurement in the VitroJet for time-efficient single particle structure determination |
| 260 | _ | _ | |a San Diego, Calif. |c 2024 |b Elsevier |
| 336 | 7 | _ | |a article |2 DRIVER |
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| 520 | _ | _ | |a Embedding biomolecules in vitreous ice of optimal thickness is critical for structure determination by cryoelectronmicroscopy. Ice thickness assessment and selection of suitable holes for data collection are currentlypart of time-consuming preparatory routines performed on expensive electron microscopes. To address thischallenge, a routine has been developed to measure ice thickness during sample preparation using an opticalcamera integrated in the VitroJet. This method allows to estimate the ice thickness with an error below ±20 nmfor ice layers in the range of 0–70 nm. Additionally, we characterized the influence of pin printing parametersand found that the median ice thickness can be reproduced with a standard deviation below ±11 nm forthicknesses up to 75 nm. Therefore, the ice thickness of buffer-suspended holes on an EM grid can be tuned andmeasured within the working range relevant for single particle cryo-EM. Single particle structures of apoferritinwere determined at two distinct thicknesses of 30 nm and 70 nm. These reconstructions demonstrate theimportance of ice thickness for time-efficient cryo-EM structure determination. |
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| 700 | 1 | _ | |a Schotman, Maaike J. G. |0 P:(DE-HGF)0 |b 1 |
| 700 | 1 | _ | |a Shahzad, Saba |0 P:(DE-Juel1)194807 |b 2 |u fzj |
| 700 | 1 | _ | |a Fromm, Simon A. |0 0000-0002-2094-1911 |b 3 |
| 700 | 1 | _ | |a Mann, Daniel |0 P:(DE-Juel1)179550 |b 4 |
| 700 | 1 | _ | |a Hennies, Julian |0 0000-0002-0555-151X |b 5 |
| 700 | 1 | _ | |a Heidler, Thomas V. |0 P:(DE-Juel1)184892 |b 6 |
| 700 | 1 | _ | |a Ashtiani, Dariush |0 P:(DE-HGF)0 |b 7 |
| 700 | 1 | _ | |a Hagen, Wim J. H. |0 0000-0001-6229-2692 |b 8 |
| 700 | 1 | _ | |a Jeurissen, Roger J. M. |0 0000-0002-1388-5691 |b 9 |
| 700 | 1 | _ | |a Mattei, Simone |0 P:(DE-HGF)0 |b 10 |
| 700 | 1 | _ | |a Peters, Peter J. |0 P:(DE-Juel1)131901 |b 11 |
| 700 | 1 | _ | |a Sachse, Carsten |0 P:(DE-Juel1)173949 |b 12 |u fzj |
| 700 | 1 | _ | |a Beulen, Bart W. A. M. M. |0 P:(DE-HGF)0 |b 13 |e Corresponding author |
| 773 | _ | _ | |a 10.1016/j.jsb.2024.108139 |g Vol. 216, no. 4, p. 108139 - |0 PERI:(DE-600)1469822-5 |n 4 |p 108139 - |t Journal of structural biology |v 216 |y 2024 |x 1047-8477 |
| 856 | 4 | _ | |u https://juser.fz-juelich.de/record/1032201/files/Ice%20thickness%20control%20and%20measurement%20control%20in%20the%20VitroJet.pdf |y OpenAccess |
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