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001032201 1001_ $$00009-0005-1709-7587$$aHenderikx, Rene J. M.$$b0$$eCorresponding author
001032201 245__ $$aIce thickness control and measurement in the VitroJet for time-efficient single particle structure determination
001032201 260__ $$aSan Diego, Calif.$$bElsevier$$c2024
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001032201 520__ $$aEmbedding biomolecules in vitreous ice of optimal thickness is critical for structure determination by cryoelectronmicroscopy. Ice thickness assessment and selection of suitable holes for data collection are currentlypart of time-consuming preparatory routines performed on expensive electron microscopes. To address thischallenge, a routine has been developed to measure ice thickness during sample preparation using an opticalcamera integrated in the VitroJet. This method allows to estimate the ice thickness with an error below ±20 nmfor ice layers in the range of 0–70 nm. Additionally, we characterized the influence of pin printing parametersand found that the median ice thickness can be reproduced with a standard deviation below ±11 nm forthicknesses up to 75 nm. Therefore, the ice thickness of buffer-suspended holes on an EM grid can be tuned andmeasured within the working range relevant for single particle cryo-EM. Single particle structures of apoferritinwere determined at two distinct thicknesses of 30 nm and 70 nm. These reconstructions demonstrate theimportance of ice thickness for time-efficient cryo-EM structure determination.
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001032201 7001_ $$0P:(DE-HGF)0$$aSchotman, Maaike J. G.$$b1
001032201 7001_ $$0P:(DE-Juel1)194807$$aShahzad, Saba$$b2$$ufzj
001032201 7001_ $$00000-0002-2094-1911$$aFromm, Simon A.$$b3
001032201 7001_ $$0P:(DE-Juel1)179550$$aMann, Daniel$$b4
001032201 7001_ $$00000-0002-0555-151X$$aHennies, Julian$$b5
001032201 7001_ $$0P:(DE-Juel1)184892$$aHeidler, Thomas V.$$b6
001032201 7001_ $$0P:(DE-HGF)0$$aAshtiani, Dariush$$b7
001032201 7001_ $$00000-0001-6229-2692$$aHagen, Wim J. H.$$b8
001032201 7001_ $$00000-0002-1388-5691$$aJeurissen, Roger J. M.$$b9
001032201 7001_ $$0P:(DE-HGF)0$$aMattei, Simone$$b10
001032201 7001_ $$0P:(DE-Juel1)131901$$aPeters, Peter J.$$b11
001032201 7001_ $$0P:(DE-Juel1)173949$$aSachse, Carsten$$b12$$ufzj
001032201 7001_ $$0P:(DE-HGF)0$$aBeulen, Bart W. A. M. M.$$b13$$eCorresponding author
001032201 773__ $$0PERI:(DE-600)1469822-5$$a10.1016/j.jsb.2024.108139$$gVol. 216, no. 4, p. 108139 -$$n4$$p108139 -$$tJournal of structural biology$$v216$$x1047-8477$$y2024
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