Single-particle cryoEM has made visualization of biological molecules and their complexes at high resolution possible without the need for crystallization. The last decade has seen a rapid growth in the number of structures determined by cryoEM; for membrane proteins, the total number of cryoEM structures has exceeded that determined by crystallography.

In this issue of IUCrJ, Subramaniam, Kühlbrandt & Henderson present an overview of the remarkable progress that has been made in electron cryo-microscopy and electron cryo-tomography.

The increasing democratization and implementation of electron cryo-microscopy appears poised to drive a new revolution in digital structural biology.

Advances in cryo-electron microscopy have enabled molecular insights into the assembly and regulation of the complement membrane attack complex.

Recent advances in single-particle cryo-electron microscopy have enabled a deeper understanding of the structural diversity and higher-order organization of aqua­porin complexes. These insights have important implications for disease biology and drug discovery.

Two recent cryogenic electron microscopy (cryoEM) structures provide a unique opportunity to compare a bacteriophage and a bacteriocin targeted at the same host. CryoEM reveals the most detailed view of contractile injection systems interacting with Clostridioides difficile.

We use sub-2 Å single-particle analysis and analytical contrast transfer function modeling to show that the Volta phase plate (VPP) boosts low-frequency signal relevant for cryo-electron tomography (cryo-ET) tilt-series alignment, improving contrast and alignment robustness in thick, crowded samples. Despite reduced subtomogram averaging resolution, these results establish the VPP's value for cryo-ET in thick, dense specimens, where tilt-series alignment is challenging and moderate-resolution information can still provide valuable biological insights.

An atomic resolution of 1.24 Å was achieved on an upgraded 200 kV electron microscope featuring a cold field emission gun, a high-resolution objective lens polepiece and an energy filter. These com­ponents transform the instrument into a cost-effective single-particle cryo-EM platform with performance com­parable to that of significantly more expensive 300 kV systems.

We present a compact mathematical framework for ab initio single-particle 3D reconstruction that reformulates the inverse problem in polar Fourier coordinates, enabling direct orientation recovery from extremely noisy 2D projection images without explicit 3D density reconstruction.

Our studies show that cryo-EM structure-guided optimization to eliminate the off-target liability of CB-5083 yields a potent and selective inhibitor of the AAA ATPase VCP/p97, a cancer target.

High-resolution cryoEM structures of MAT enzyme complexes help to explain why MATα1 selectively forms a stable complex with MATβV1 but not with MATβV2, despite sharing high sequence and structural identity with MATα2.

We introduce AlignPCA-2D, a PCA-space Euclidean alignment method for rapid and interpretable 2D classification of cryo-EM particle images. The approach preserves essential structural variability while greatly reducing computational cost, offering a lightweight alternative to existing large-scale cryo-EM classification pipelines.

Analysis of SARS-CoV-2 spike protein heterogeneity using cryo-EM data reveals cooperativity between receptor-binding domains.

We present an integrated sample preparation workflow that combines photocaged ligand activation and controlled triggering, coupled with rapid vitrification to enable time-resolved cryoET studies on bacterial chemotaxis at millisecond timescales.