WikiSysop: Created page with "== Citation == Aiyer, Sriram / Baldwin, Philip R. / Tan, Shi Min / Shan, Zelin / Oh, Juntaek / Mehrani, Atousa / Bowman, Marianne E. / Louie, Gordon / Passos, Dario Oliveira / Đorđević-Marquardt, Selena / others. Overcoming resolution attenuation during tilted cryo-EM data collection. 2024. Nature communications, Vol. 15, No. 1, p. 389 == Abstract == Structural biology efforts using cryogenic electron microscopy are frequently stifled by specimens adopting “prefe..."

2024-08-20T06:40:28Z

Created page with "== Citation == Aiyer, Sriram / Baldwin, Philip R. / Tan, Shi Min / Shan, Zelin / Oh, Juntaek / Mehrani, Atousa / Bowman, Marianne E. / Louie, Gordon / Passos, Dario Oliveira / Đorđević-Marquardt, Selena / others. Overcoming resolution attenuation during tilted cryo-EM data collection. 2024. Nature communications, Vol. 15, No. 1, p. 389 == Abstract == Structural biology efforts using cryogenic electron microscopy are frequently stifled by specimens adopting “prefe..."

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== Citation ==

Aiyer, Sriram / Baldwin, Philip R. / Tan, Shi Min / Shan, Zelin / Oh, Juntaek / Mehrani, Atousa / Bowman, Marianne E. / Louie, Gordon / Passos, Dario Oliveira / Đorđević-Marquardt, Selena / others. Overcoming resolution attenuation during tilted cryo-EM data collection. 2024. Nature communications, Vol. 15, No. 1, p. 389

== Abstract ==

Structural biology efforts using cryogenic electron microscopy are frequently
stifled by specimens adopting “preferred orientations” on grids, leading to
anisotropic map resolution and impeding structure determination. Tilting the
specimen stage during data collection is a generalizable solution but has historically
led to substantial resolution attenuation. Here, we develop updated
data collection and image processing workflows and demonstrate, using
multiple specimens, that resolution attenuation is negligible or significantly
reduced across tilt angles. Reconstructions with and without the stage tilted as
high as 60° are virtually indistinguishable. These strategies allowed the
reconstruction to 3 Å resolution of a bacterial RNA polymerase with preferred
orientation, containing an unnatural nucleotide for studying novel base pair
recognition. Furthermore, we present a quantitative framework that allows
cryo-EM practitioners to define an optimal tilt angle during data acquisition.
These results reinforce the utility of employing stage tilt for data collection
and provide quantitative metrics to obtain isotropic maps.

== Keywords ==

== Links ==

https://www.nature.com/articles/s41467-023-44555-7

== Related software ==

== Related methods ==

== Comments ==

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