Data Processing Tutorials
Tutorials on how to use CryoSPARC jobs and processing cryo-EM data.
Case Studies

Processing EMPIAR-10739 including handling global pseudosymmetry, using 3DVA to guide classification strategies, separating low population classes, and local refinement of a flexible region.

Processing EMPIAR-10853 including Micrograph Denoising, Micrograph Junk Detector, Subset Particles by Statistic, Local Refinement and focussed 3D Classification to improve ligand density.

Processing EMPIAR-10059 in CryoSPARC v4.6 with a focus on particle curation, developing an intuition for when a domain may be blurred due to flexibility, and how to handle this type of flexibility.

Processing EMPIAR-10256 with a focus on handling pseudosymmetry and 3D Classification parameter choices.

Processing EMPIAR-10668 including Local Refinement, 3D Variability Analysis, 3D Flex and classification of continuous heterogeneity.

Processing EMPIAR-10716 with a focus on high-symmetry ab-initio reconstruction, local symmetry, non-point-group symmetry, symmetry expansion, and custom geometry operations.

Processing a combined dataset of an ion channel with a focus on techniques for separating conformational states from a heterogeneous mixture.

An analysis of the impact of data collection and particle picking techniques on orientation bias, including strategies for improving map quality and isotropy.


Friendly, approachable introductions to cryoEM data processing in CryoSPARC with a focus on the exploratory, "try-it-and-see" nature of single-particle analysis. Kindly reproduced from Oliver Clarke, PhD, Assistant Professor of Physiology and Cellular Biophysics at Columbia University.


A detailed description of the common plots that CryoSPARC makes across multiple job types.
Data Processing
Alongside these tutorials, see: All Job Types in CryoSPARC and Tutorial Videos
Files, Data and Import
Tutorial: Negative Stain DataTutorial: Phase Plate DataTutorial: EER File SupportTutorial: EPU AFIS Beam Shift ImportPreprocessing
Tutorial: Patch Motion and Patch CTFTutorial: Float16 SupportParticle Picking
Tutorial: Particle Picking CalibrationTutorial: Blob Picker TunerHelical Reconstruction
Tutorial: Helical Processing using EMPIAR-10031 (MAVS)3D Refinement
Tutorial: Maximum Box Sizes for RefinementTutorial: CTF RefinementTutorial: Ewald Sphere CorrectionTutorial: Symmetry RelaxationTutorial: Orientation DiagnosticsTutorial: BILD files in CryoSPARC v4.4+Local Refinement
Tutorial: Mask CreationConformational Variability
Tutorial: 3D ClassificationTutorial: 3D Variability Analysis (Part One)Tutorial: 3D Variability Analysis (Part Two)Tutorial: 3D Flexible RefinementTutorial: 3D Flex Mesh PreparationLast updated
