EMD-29573

Single-particle
2.76 Å
EMD-29573 Deposition: 26/01/2023
Map released: 22/02/2023
Last modified: 22/02/2023
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links

EMD-29573

Mouse apoferritin from data collected on Talos Arctica microscope equipped with a K3 camera operating in counting mode, in 0-50 nm ice thickness

EMD-29573

Single-particle
2.76 Å
EMD-29573 Deposition: 26/01/2023
Map released: 22/02/2023
Last modified: 22/02/2023
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
Sample Organism: Mus musculus
Sample: apoferritin
Raw data: EMPIAR-11397

Deposition Authors: Neselu K , Wang B , Rice WJ , Potter CS , Carragher B , Chua EYD
Measuring the effects of ice thickness on resolution in single particle cryo-EM.
Neselu K , Wang B , Rice WJ , Potter CS , Carragher B , Chua EYD
(2023) J Struct Biol X , 7 , 100085 - 100085
PUBMED: 36742017
DOI: doi:10.1016/j.yjsbx.2023.100085
ISSN: 2590-1524
Abstract:
Ice thickness is a critical parameter in single particle cryo-EM - too thin ice can break during imaging or exclude the sample of interest, while ice that is too thick contributes to more inelastic scattering that precludes obtaining high resolution reconstructions. Here we present the practical effects of ice thickness on resolution, and the influence of energy filters, accelerating voltage, or detector mode. We collected apoferritin data with a wide range of ice thicknesses on three microscopes with different instrumentation and settings. We show that on a 300 kV microscope, using a 20 eV energy filter slit has a greater effect on improving resolution in thicker ice; that operating at 300 kV instead of 200 kV accelerating voltage provides significant resolution improvements at an ice thickness above 150 nm; and that on a 200 kV microscope using a detector operating in super resolution mode enables good reconstructions for up to 200 nm ice thickness, while collecting in counting instead of linear mode leads to improvements in resolution for ice of 50-150 nm thickness. Our findings can serve as a guide for users seeking to optimize data collection or sample preparation routines for both single particle and in situ cryo-EM. We note that most in situ data collection is done on samples in a range of ice thickness above 150 nm so these results may be especially relevant to that community.