Rapid freezing of cells can provide outstanding structure preservation and time resolution of dynamic cellular processes. Electron tomography of rapidly frozen specimens (cryo-ET) is a powerful technique for imaging biological structures in their native state and in an unperturbed cellular environment. We integrate high resolution imaging by either cryo-ET and sub-tomogram averaging or TYGRESS (Tomography-Guided 3D Reconstruction of Subcellular Structures), with comparative genetics, biochemical methods and EM-visible labeling to deconstruct the in situ 3D structure and functional organization of macromolecular complexes. Among different model systems, we use e.g. cilia and flagella to advance techniques and approaches for high-resolution imaging of complex cellular structures. Cilia and flagella are conserved and ubiquitous eukaryotic organelles that are composed of more than 600 different proteins and have important biological roles in motility and sensation; defects in their assembly or function cause severe human diseases. Our cryo-ET studies visualize the three-dimensional structures of intact wild-type and mutant flagella, and dissect the organization of key macromolecular complexes in different functional states. Such information can provide detailed insights into the structural basis and ultimately the function of many cellular processes.

Studying the molecular organization and function of cells using cryo-electron tomography

2019 Fall National Meeting

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