Hi Vincent,
It is important that the greyscale values in the movies are the same as
in the average micrographs used in refinement. What I mean is that the
sum of all movie-frames (divided by the number of frames) should give
the same image as the average micrograph.
And yes, image_handler can do this.
HTH,
S
On 05/06/2015 04:32 PM, Vincent Yip wrote:
> Dear all,
>
> When I collected movie data, the movie was only dark-subtracted and not
> gain-corrected by our image-collecting software in order to save hard drive
> space. The software also spits out the gain reference maps so that the gain
> can be corrected during the movie-frame averaging and alignment step (using
> a home-made script).
>
> I have used these averaged micrographs for all the processing steps to
> 3D-auto-refine in Relion. To further improve the reconstruction, I have
> decided to do post-processing with movie alignment with individual
> particles (particle polishing). I am wondering if I can use my
> dark-subtracted movies for this step or if I need to get a gain-corrected
> movie stack. If I will indeed need a gain-corrected movie stack, can
> relion_image_handler do that? Or do I need to use other packages?
>
> Thank you very much for you help!
>
> Vincent
>
--
Sjors Scheres
MRC Laboratory of Molecular Biology
Francis Crick Avenue, Cambridge Biomedical Campus
Cambridge CB2 0QH, U.K.
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