Multicolor single particle reconstruction of protein complexes

Single-particle reconstruction (SPR) from electron microscopy images is widely used in structural biology, but lacks direct information on protein identity. To address this limitation, we developed a computational and analytical framework that reconstructs and co-aligns multiple proteins from 2D superresolution fluorescence images. We demonstrate our method by generating multicolor 3D reconstructions of several proteins within the human centriole, revealing their relative locations, dimensions and orientations.