Detalle de: AlignPCA -2 D : PCA-reduced Euclidean vector alignment for 2D classification in cryo-EM

Cryogenic electron microscopy (cryo-EM) has transformed structural biology by enabling the high-resolution reconstruction of macromolecular complexes from noisy projection images. However, the intrinsic heterogeneity and low signal-to-noise ratio of cryo-EM datasets make 2D classification a critical and computationally demanding step in the processing workflow. Here, we introduce AlignPCA -2 D , a principal component analysis (PCA)-space Euclidean vector alignment method for fast, interpretable 2D classification in cryo-EM. By projecting particle images and class representations into a compressed latent PCA space, AlignPCA -2 D reduces data dimensionality while preserving meaningful structural variability. The image-to-class assignment is then performed using Euclidean distance, enabling efficient and accurate classification. We benchmark AlignPCA -2 D against established cryo-EM software, such as RELION and CryoSPARC , and demonstrate that it achieves competitive alignment accuracy while substantially reducing computational cost. This approach provides a lightweight alternative for large-scale 2D classification tasks, and its modular design makes it compatible with existing cryo-EM processing pipelines.