%0 Journal Article
%A Henderikx, Rene J. M.
%A Schotman, Maaike J. G.
%A Shahzad, Saba
%A Fromm, Simon A.
%A Mann, Daniel
%A Hennies, Julian
%A Heidler, Thomas V.
%A Ashtiani, Dariush
%A Hagen, Wim J. H.
%A Jeurissen, Roger J. M.
%A Mattei, Simone
%A Peters, Peter J.
%A Sachse, Carsten
%A Beulen, Bart W. A. M. M.
%T Ice thickness control and measurement in the VitroJet for time-efficient single particle structure determination
%J Journal of structural biology
%V 216
%N 4
%@ 1047-8477
%C San Diego, Calif.
%I Elsevier
%M FZJ-2024-06070
%P 108139 -
%D 2024
%X 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.
%F PUB:(DE-HGF)16
%9 Journal Article
%$ 39433138
%U <Go to ISI:>//WOS:001343861100001
%R 10.1016/j.jsb.2024.108139
%U https://juser.fz-juelich.de/record/1032201