Streaming heterogeneous refinement (feature request)

CryoSPARC Live
1

Hi,

For many membrane proteins, particularly in nanodisc, it is often necessary to perform at least one round of 3D cleanup to get a decent resolution reconstruction.

While this is sort-of possible in Live using multi class ab initio, the Streaming Refinement module as far as I can tell takes the selected particles from 2D, not the ones from ab initio (which makes sense seeing as ab initio is not set up for streaming).

Would it be possible to add in a streaming heterogeneous refinement module, to allow for continuous cleanup in 3D (and potentially separation of different conformations)?

Cheers
Oli

2

Would still love a streaming hetero refinement module in Live, or streaming ab initio I guess - some means of on the fly refinement of a selected 3D class. Right now with a complex sample I can often separate multiple components by ab initio, but I would like to be able to quickly refine a single class so I can know the composition (and whether or not further collection will be useful) - it would greatly improve the usefulness of live for complex samples (e.g. allowing on the fly separation of complex from residual free nanodisc)

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3

I would second this request for streaming hetero refinement, this would be an excellent feature to have. We typically always also do a 3D cleanup before we are able to reach a reasonable resolution also with membrane proteins.

4

Would still really love to see a Live version of heterogenous refinement!

As we increasingly deal with more heterogeneous samples (e.g. partially purified native cell extracts), some form of on-the fly classification & refinement will become ever more useful.

Cheers
Oli

5

Bumping this :slight_smile: because we would still love to see this. We basically do all of our initial sorting now using heterogenous refinement and only use 2D in later stages of sorting to pick out classes with high res features. The goal would be to have this running on our live session such that we would have a really good stack of particles that people could take to refinement immediately after data collection.

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