TY  - JOUR
AU  - Henderikx, Rene J. M.
AU  - Schotman, Maaike J. G.
AU  - Shahzad, Saba
AU  - Fromm, Simon A.
AU  - Mann, Daniel
AU  - Hennies, Julian
AU  - Heidler, Thomas V.
AU  - Ashtiani, Dariush
AU  - Hagen, Wim J. H.
AU  - Jeurissen, Roger J. M.
AU  - Mattei, Simone
AU  - Peters, Peter J.
AU  - Sachse, Carsten
AU  - Beulen, Bart W. A. M. M.
TI  - Ice thickness control and measurement in the VitroJet for time-efficient single particle structure determination
JO  - Journal of structural biology
VL  - 216
IS  - 4
SN  - 1047-8477
CY  - San Diego, Calif.
PB  - Elsevier
M1  - FZJ-2024-06070
SP  - 108139 -
PY  - 2024
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - 39433138
UR  - <Go to ISI:>//WOS:001343861100001
DO  - DOI:10.1016/j.jsb.2024.108139
UR  - https://juser.fz-juelich.de/record/1032201
ER  -