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@ARTICLE{Henderikx:1032201,
author = {Henderikx, Rene J. M. and Schotman, Maaike J. G. and
Shahzad, Saba and Fromm, Simon A. and Mann, Daniel and
Hennies, Julian and Heidler, Thomas V. and Ashtiani, Dariush
and Hagen, Wim J. H. and Jeurissen, Roger J. M. and Mattei,
Simone and Peters, Peter J. and Sachse, Carsten and Beulen,
Bart W. A. M. M.},
title = {{I}ce thickness control and measurement in the {V}itro{J}et
for time-efficient single particle structure determination},
journal = {Journal of structural biology},
volume = {216},
number = {4},
issn = {1047-8477},
address = {San Diego, Calif.},
publisher = {Elsevier},
reportid = {FZJ-2024-06070},
pages = {108139 -},
year = {2024},
abstract = {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.},
cin = {ER-C-3},
ddc = {540},
cid = {I:(DE-Juel1)ER-C-3-20170113},
pnm = {5352 - Understanding the Functionality of Soft Matter and
Biomolecular Systems (POF4-535) / 5241 - Molecular
Information Processing in Cellular Systems (POF4-524)},
pid = {G:(DE-HGF)POF4-5352 / G:(DE-HGF)POF4-5241},
typ = {PUB:(DE-HGF)16},
pubmed = {39433138},
UT = {WOS:001343861100001},
doi = {10.1016/j.jsb.2024.108139},
url = {https://juser.fz-juelich.de/record/1032201},
}