Helical Refine (refinement not converging in the 1st attempt)

Dear cryoSPARC team

We’ve been enjoyed the helical refinement function in cryoSPARC. But recently I noticed a strange behavior of the Helical refinement from two unrelated helical objects. They are both pretty large in diameter (250-350 Å), and the tube can be viewed as a thin shell.

What I found is that the refinement didn’t converge in the first Helical refinement job with the correct symmetry (tiny search range for the rise and twist because it is calculated from layer lines at high resolution). However, if I copied this Helical refinement job, using exactly the same parameters and also using the output volume of the 1st job as input, the 2nd refinement will converge. Any thoughts? Currently, it didn’t create any problem for me, but one can imagine it may be an issue when one has 100 possible helical symmetries to test.

1st job (reported ~10Å)

2nd job (still going, ~3.3Å)


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Dear @Jerry,

Thanks for the feedback! Reports like this help us improve the robustness of the algorithms we implement :slight_smile:

We have noticed that this can sometimes happen on the datasets available to us, and most often the issue was related to both half maps diverging from each other early on (usually through slight differences in rotation/orientation). This causes the FSCs to be incorrect, sometimes showing a lot of artefacts and spikes. The typical way to work around this is to average the two half-maps together at low resolution only, and by default, this is done at resolution of 20Å or below. Though for objects with helical symmetry and smaller asymmetric units, it seems 20Å might not always be sufficient, as more of the signal used to align particles to the volume is moderate or higher frequency. E.g. in this dataset, at ~10Å it looks like you can just see the asymmetric unit, but I imagine at 20Å it might just look like a featureless cylinder.

Could you try changing the GSFSC split resolution (A) parameter to be a bit lower in value, say 12-15Å, to see if this alleviates the issue in the first job? As well, do you notice that the FSCs in the first job contain lots of spikes or artefacts at resolutions beyond 20Å?

Something else that may be related to this is the initialization. In the first job you ran (top picture), did you pass in an initial model, or did you let the refinement generate one itself? In my experience the problem mentioned above is more common when the initial model is generated from scratch, possibly because we’re starting “further away” from the optimal structure, so there’s more ways in which the half maps could diverge.


Thank you Michael for the useful tip. I tried 2 things:

(1) copied the 1st job, and changed GSFSC split resolution (A) to 12Å. And it worked (reported ~3.3Å resolution)!

(2) copied the 1st job, and deleted the starting volume (what I used for the starting volume is a cylinder shell generated according to its diameter. It had a bug without starting volume, but that’s another story. See my previous post). It reported ~8Å.

So it seems the GSFSC split resolution (A) is really the solution, and I will set that to 12Å by default in future helical projects. Maybe you guys may consider this as well in your future release?


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