Narrow-beam geometry improves the efficiency of cryo-EM

Authors

Matinyan, S., Filipcik, P., Genderen, E. v., Abrahams, J. P.

Abstract

Cryo-electron microscopy (cryo-EM) of biological specimens is limited by radiation damage and a low signal-to-noise ratio (SNR). Here, we show that reducing the illuminated area substantially slows the observed diffraction decay in protein microcrystals. We further show that narrow parallel-beam electron diffraction from thin non-crystalline biological specimens provides substantially higher reciprocal-space SNR than conventional cryo-EM imaging. We developed a multimodal scanning workflow, 4D-para-STEM, that records narrow-beam diffraction patterns together with corresponding images. Using viruses, peptide assemblies, and microtubules, we demonstrate interpretable diffraction signals from both crystalline and non-crystalline biological specimens. Together, these results show that narrow parallel-beam scanning reduces observed radiation damage and improves the SNR in cryo-EM.

Preprint server: bioRxiv
The authors list and abstract were imported from bioRxiv on 09 Jul 2026.

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