Explore a cutting-edge approach to recovering three-dimensional atomic structures of flexible macromolecules from heterogeneous single-particle cryo-electron microscopy (cryo-EM) datasets in this 51-minute conference talk. Delve into the innovative method that combines prior biological knowledge with cryo-EM images to determine atomic structure deformations. Learn how the space of possible atomic structures is parametrized using biological knowledge, and how conformation parameters are estimated through a linear combination of leading eigenvectors from a graph Laplacian. Discover how this technique approximates the spectral properties of the macromolecule's conformation manifold, offering new insights into structural biology and cryo-EM analysis.