Hello CryoSPARC users! The Stanford-SLAC CryoEM Center (S2C2) holds annual image processing workshops, and we were generously invited to present at this year’s session. We have posted the CryoSPARC sessions to YouTube, slightly edited to remove gaps in the recordings. We believe the topics covered in these lectures will be interesting to all users, but they are especially targeted toward relatively new cryo-EM practitioners, covering foundational concepts in the theory and practice of cryo-EM image processing.
The first video in the series introduces the format of this workshop and presents an overview of how we collect cryo-EM data and various difficulties we must overcome during image processing.
The second video presents a “standard workflow” using TRPV1, meant to be a useful starting workflow for most single-particle datasets. Of course, there are many alternate workflows which will be just as effective!
The third video covers important considerations and potential pitfalls when working with symmetric samples, using the TRPV5 ion channel as an example.
In the fourth video, we discuss how we processed an Encapsulated Ferritin sample with non-point-group symmetry to high resolution, including using several different jobs to perform a “custom symmetry expansion”.
The fifth video presents the benefits and drawbacks of various methods of handling discrete heterogeneity, using a FaNaC1 dataset as an example.
The final video discusses the two main means of working with continuous heterogeneity in CryoSPARC (namely, 3D Variability Analysis and 3D Flexible Refinement). This video covers both the theoretical workings of these jobs, as well as considerations for inputs and parameter choices.
We are very grateful for S2C2’s invitation to present at this workshop, and encourage users to check out the other speakers’ talks and those from previous years on S2C2’s website!