- EMDB-29594: Mouse apoferritin from data collected on Talos Arctica microscope... -
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Database: EMDB / ID: EMD-29594
Title
Mouse apoferritin from data collected on Talos Arctica microscope equipped with a K3 camera operating in super resolution mode, in 200-500 nm ice thicknessFerritin
Map data
Mouse apoferritin from data collected on Talos Arctica microscope equipped with a K3 camera operating in super resolution mode, in 200-500 nm ice thicknessFerritin
Sample
Complex: apoferritinFerritin
Keywords
Mouse apoferritin / Talos Arctica / K3 camera / super resolution mode / 200-500 nm ice thickness / METAL BINDING PROTEIN
Biological species
Mus musculus (house mouse)
Method
single particle reconstruction / cryo EM / Resolution: 8.82 Å
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
GM103310
United States
Citation
Journal: J Struct Biol X / Year: 2023 Title: Measuring the effects of ice thickness on resolution in single particle cryo-EM. Authors: Kasahun Neselu / Bing Wang / William J Rice / Clinton S Potter / Bridget Carragher / Eugene Y D Chua / 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 ...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.
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Annotation
Mouse apoferritin from data collected on Talos Arctica microscope equipped with a K3 camera operating in super resolution mode, in 200-500 nm ice thickness
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