User Manual for Tomography-Guided 3D Reconstruction of Subcellular Structures (TYGRESS)

Abstract Recent advances in cryo-electron microscopy (cryo-EM) are paving the way to determining isolated three-dimensional (3D) macromolecular structures at near-atomic resolution using single-particle cryo-electron microscopy (SP-cryo-EM). However, determining the subcellular structures in intact cells and organelles using cryo-electron tomography (cryo-ET) and subtomogram averaging, another cryo-EM technique, with comparable resolution remains a challenge. Current methodologies can only reach a resolution of several nanometers in most samples studied. Here, we introduce a new hybrid method, called Tomography-Guided 3D Reconstruction of Subcellular Structures (TYGRESS) that is able to achieve structural determination of subcellular structures within their natural crowded environment with nanometer-resolution by combining the advantages of cryo-ET and SP-cryo-EM.