Hi,
I’m working with a membrane protein that shows some flexibility induced loss of feature in some regions.
Because of that I was thinking to collect a few tilt-pair movies, to get more thorough understanding about the features of the particles.
However, I was wondering how to treat a tilt-pair data in cryosparc. Unlike EMAN2, cryosparc does not have any tilt pair boxer program.
Some suggestions on this matter would be very helpful.
Thank you
Abhisek
Hi @diffracteD,
Thanks for your post. Unfortunately we have never tried to process or support tilt-pair data acquisition - as far as I’m aware it is relatively rarely used.
How would you expect the tilt pair data to help with the flexibility-induced loss of features?
why don’t you pick in EMAN2 and then import to CS, e.g extract in RELION3 and then import stack in CS?
Depending on flexibility, you would get higher resolution with flexibility analysis of normal SPA dataset rather than tilt pairs.
Yes sure. I was skeptical about recent microscope time I have so I thought to collect a small set with tilt pair.
But now the time issue is gone, I’m doing SPA.
The particle contrast I’m working with is very low (the particles stays nice in thicker ice only). CS does a great job picking low contrast particles, however, having a hard time getting internal features.
try denoising with Janni or Topaz.
if it is maybe in micelle/nanodisc, you might want to try partial signal subtraction followed if necessary by multibody refinement: though the latter might be tricker since protein is small.