movie processing

[patrick bron]

Dear all 

I have recorded images on a Titan Krios with a Flacon 2 camera using the movie mode. After a few cycles, I obtained a first 3D structure at 4.7A resolution using particles extracted from the final image recorded after 1 sec exposition.

I would like to know if you have developed some procedures for Auto3dem in order to include particles from video frames rather than particles from the total exposure image ?

As aligning particles together from selected frames and merging them ? or using and processing independently particles from selected frames ? …..

Sincerely 

Patrick 

Patrick Bron, PhD, Team leader

Team "Multi-Scale Structural Biology"

« Structural Biology » Department

Centre de Biochimie Structurale

INSERM U1054 / CNRS UMR5048

29 rue de Navacelles

34090 Montpellier, France

Phone: 33 4 67 41 77 29

Fax: 33 4 67 41 79 12

http://www.cbs.cnrs.fr

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Passerelle antivirus CBS

[Giovanni Cardone]

Hi Patrick,

at the moment Auto3DEM accepts only micrographs as input, and not frames, since it does not include any procedure for generating drift-corrected, sub-averages of frames from a DDD camera, although it is under consideration.
This means you have to rely on external programs, in some cases provided by the camera manufacturer, to get different subsets of frames to form the input micrographs.
However, once you have these new images, you can repeat the boxing procedure using all the information derived from the original images, and save the new stacks in a directory that is different from the previous one. At this point it is just a matter to make the parameters (.dat) files of your last iteration to point to this new directory (autopp option I), and you can continue the processing using the same orientation and alignment parameters but with the new images.
If you need any help with the procedure, just let me know.

Best,
Giovanni

Passerelle antivirus CBS --

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-- 
Giovanni Cardone
Baker's Laboratory
University of California, San Diego
Department of Chemistry and Biochemistry
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