Unblur & Summovie

Hi,

I have a FalconII movie data. total dose: 40 e/A2 in 30 frames.

I have used the unblur to align and exposure filter the frames with the default parameters (Bfactor of 1500). I have calculated the power specturm of the averaged (summed) images before and after the unblur filtering.I see that the high resolution Thon-rings are more visible for the original image than for the exposure-filtered image. Is it expected like this?. Please help me to understand this.

Dear all,

I am wondering what the "restore noise power after filtering" function actually means, as I could not find a description in the eLife paper. By default this option is set to yes in unblur and sum_movie. What is it good for and in which situations should it be used and in which situations rahter not?

Thanks again for any help and best resgards
Niels

Dear all,
I have a question how to deal with image stacks which start after a pre-exposure, i.e. the first frame aquired by the camera is not included in the saved stack.

Hi Tim,

Very quick question, I am just wondering whether there is a way to bin data during Unblur processing ? or I just have to bin data separately after Unblur.

Best
Hun

Hi Tim-

Quick question: when unblur outputs an average image, which frame does it use as the reference for the alignment? I.e. is it the first frame, middle frame, etc.?

Thanks,
-Greg Alushin

Hi Tim,
I am wondering if there is a way to apply the gain (dm4) to gain-uncorrected-compressed-TIF file?
Where can I set the CPU number for processing?
Is it possible to write out intermediate corrected stack, so we can use local motion correction and then apply exposure filtering?
Thanks,
Lei

I'm a little confused by the parameter of "Exposure per frame". I initially thought that the exposure (e-/A^2) referred to the dose received by the specimen. But the method part of the elife paper (eLife 2015;10.7554/eLife.06980) says "The exposure per frame as reported by Digital Micrograph (Gatan, Inc.) was 0.769 e-/Å2 which corresponds to an exposure of 8 electrons/pixel/s on the camera." When I used 8e-/pix/s to calculate the total exposure, it gave me 100e-/A^2. This means the value of 100 corresponds to the dose on the camera.

Hi,

I wanted to give a go at unblur and summovie.
I'm unsure how to launch them on a folder full of mrc stacks?
I guess I should create a .com file very much like rmeasure is using but I'm not sure what it should contain.
Could you give me an example of how to do that?

Thank you,
Ludo