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-Structure paper
| タイトル | Measuring the effects of ice thickness on resolution in single particle cryo-EM. |
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| ジャーナル・号・ページ | J Struct Biol X, Vol. 7, Page 100085, Year 2023 |
| 掲載日 | 2023年1月24日 |
 著者 | Kasahun Neselu / Bing Wang / William J Rice / Clinton S Potter / Bridget Carragher / Eugene Y D Chua /  |
| PubMed 要旨 | 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. |
 リンク | J Struct Biol X / PubMed:36742017 / PubMed Central |
| 手法 | EM (単粒子) |
| 解像度 | 2.36 - 10.18 Å |
| 構造データ |  EMDB-29393: Mouse apoferritin from data collected on Titan Krios with K3 camera in counting mode with a 20 eV energy filter slit, in 200-500 nm Ice thickness  EMDB-29513: Mouse apoferritin from data collected on Titan Krios with K3 camera in counting mode with a 20 eV energy filter slit, in 150-200 nm ice thickness  EMDB-29535: Mouse apoferritin from data collected on Titan Krios with K3 camera in counting mode with a 20 eV energy filter slit, in 50-100 nm ice thickness  EMDB-29536: Mouse apoferritin from data collected on Titan Krios with K3 camera in counting mode with a 20 eV energy filter slit, in 0-50 nm ice thickness  EMDB-29554: Mouse apoferritin from data collected on Titan Krios with K3 camera in counting mode without energy filter, in 0-50 nm ice thickness  EMDB-29555: Mouse apoferritin from data collected on Titan Krios with K3 camera in counting mode without energy filter, in 50-100 nm ice thickness  EMDB-29556: Mouse apoferritin from data collected on Titan Krios with K3 camera in counting mode without energy filter, in 100-150 nm ice thickness  EMDB-29557: Mouse apoferritin from data collected on Titan Krios with K3 camera in counting mode without energy filter, in 150-200 nm ice thickness  EMDB-29558: Mouse apoferritin from data collected on Titan Krios with K3 camera in counting mode without energy filter, in 200-500 nm ice thickness  EMDB-29559: Mouse apoferritin from data collected on Titan Krios with K3 camera in counting mode with a 20 eV energy filter slit, in 100-150 nm Ice thickness  EMDB-29566: Mouse apoferritin from data collected on Glacios microscope equipped with a Falcon 3 camera operating in integrating mode, in 0-50 nm ice thickness  EMDB-29567: Mouse apoferritin from data collected on Glacios microscope equipped with a Falcon 3 camera operating in integrating mode, in 50-100 nm ice thickness  EMDB-29568: Mouse apoferritin from data collected on Glacios microscope equipped with a Falcon 3 camera operating in integrating mode, in 100-150 nm ice thickness  EMDB-29569: Mouse apoferritin from data collected on Glacios microscope equipped with a Falcon 3 camera operating in integrating mode, in 150-200 nm ice thickness  EMDB-29570: Mouse apoferritin from data collected on Glacios microscope equipped with a Falcon 3 camera operating in integrating mode, in 200-500 nm ice thickness  EMDB-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  EMDB-29574: Mouse apoferritin from data collected on Talos Arctica microscope equipped with a K3 camera operating in counting mode, in 50-100 nm ice thickness  EMDB-29575: Mouse apoferritin from data collected on Talos Arctica microscope equipped with a K3 camera operating in counting mode, in 100-150 nm ice thickness  EMDB-29576: Mouse apoferritin from data collected on Talos Arctica microscope equipped with a K3 camera operating in counting mode, in 150-200 nm ice thickness  EMDB-29577: Mouse apoferritin from data collected on Talos Arctica microscope equipped with a K3 camera operating in counting mode, in 200-500 nm ice thickness  EMDB-29589: Mouse apoferritin from data collected on Talos Arctica microscope equipped with a K3 camera operating in super resolution mode, in 0-50 nm ice thickness  EMDB-29591: Mouse apoferritin from data collected on Talos Arctica microscope equipped with a K3 camera operating in super resolution mode, in 50-100 nm ice thickness  EMDB-29592: Mouse apoferritin from data collected on Talos Arctica microscope equipped with a K3 camera operating in super resolution mode, in 100-150 nm ice thickness  EMDB-29593: Mouse apoferritin from data collected on Talos Arctica microscope equipped with a K3 camera operating in super resolution mode, in 150-200 nm ice thickness  EMDB-29594: 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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mouse (ネズミ)