CryoFSL: an annotation-efficient, few-shot learning framework for robust protein particle picking in cryo-electron microscopy micrographs.

Accurate identification of protein particles in cryo-electron microscopy (cryo-EM) micrographs is crucial for high-resolution structure determination, but remains challenging due to the heavy reliance on extensive annotated datasets and the difficulty of ensuring robustness under low signal-to-noise ratio (SNR) conditions. Current approaches require large annotations and exhibit poor generalization to new protein targets. We present CryoFSL (Cryo-EM Few Shot-Learning), a novel few-shot learning framework built on Segment Anything Model 2 with lightweight adapters, enabling robust particle picking with as few as five labeled micrographs and significantly reducing the annotation burden. The framework's hierarchical adapter design supports dynamic feature modulation for low-SNR and heterogeneous conditions, resolving the trade-off between annotation burden and performance. CryoFSL surpasses both traditional template-based methods and state-of-the-art deep learning models across diverse proteins in the few-shot learning setting, achieving superior recall, precision, and 3D reconstruction resolution with minimal supervision. It maintains stability across heterogeneous micrographs and consistently detects high-quality particles with fewer false-positives. Notably, CryoFSL achieves competitive resolution in density map reconstruction with just a fraction of the particles picked by other methods, redefining efficiency and quality in cryo-EM analysis. This work paves the way for scalable, generalizable, and annotation-efficient particle-picking pipelines. The code is available at https://github.com/biplabpoudel25/CryoFSL.