FaNaC1 and Continuous Heterogeneity (Part 6 of 6) скачать в хорошем качестве

FaNaC1 and Continuous Heterogeneity (Part 6 of 6)

In the final video (Part 6 of 6) in this cryo-EM data processing series, we consider the same FaNaC1 dataset as in Part 5, but consider the apo and ligand-bound states as existing on a continuous spectrum. We begin by considering the difficulty inherent in extracting the conformation of individual particle images, and why results from these techniques should always be validated using orthogonal, biochemical methods. We then discuss the theory and practice of 3D Variability Analysis in CryoSPARC. In 3D Variability Analysis, each particle is modeled as a consensus refinement plus some linear combination of difference volumes. This makes the technique computationally lighter-weight than alternatives and can produce good results in many situations. Finally, we discuss 3D Flexible Refinement (3DFlex). This technique directly models continuous deformation of the consensus refinement using nonlinear, machine-learning techniques. We discuss important steps in setting up 3DFlex, including mesh design and important parameter choices. Chapters: 00:00 Introduction to Continuous Heterogeneity 04:00 Can You Do Thermodynamics With Cryo-EM? 14:10 Flexible Forward Model 17:55 3D Variability Analysis - Theory 24:14 3D Variability Analysis - Making a Job 29:19 3D Variability Analysis - Interpretation 50:01 Q&A: 3D Variability Analysis 1:03:54 3D Flexible Refinement - Motivation 1:07:51 Flex Data Prep 1:08:18 Flex Mesh Prep 1:11:09 Flex Training 1:17:47 Flex Generate 1:21:06 Using 3DVA results in 3D Flex 1:22:52 Flex Reconstruct 1:26:52 Final Q&A This video is the last in a series of six recordings covering single particle cryo-EM data processing in CryoSPARC from the 2024 Single-Particle Cryo-EM Image Processing Workshop held by the Stanford-SLAC CryoEM Center (S2C2) in 2024. The original recordings are courtesy of S2C2. Captions were automatically generated by Zoom and may contain errors. Workshop: https://s2c2.slac.stanford.edu/traini... S2C2: https://s2c2.slac.stanford.edu/traini... More tutorials and videos are available in the CryoSPARC Guide: https://guide.cryosparc.com/