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<p class="MsoNormal">Hi there<o:p></o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal"><span lang="EN-US">I encountered a weird problem with Relion 4 polishing in a dataset with 10 subsets of data from a Falcon 4i camera (EER movies).<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Each subset was measured with the same total exposure of 50 e-/A^2, but the exposure time per micrograph to achieve that was significantly different in some subsets.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Therefore, each subset has a different amount of EER fractions ( in the range of 600 – 1200 fractions).
<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Since in relion one can only choose how many EER fractions are combined to one frame, the amount of frames after EER fraction combination varies from 49 to 51.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">I figured out that with the –find-shortest option, a polishing job on all the datasets combined would find out that 49 is the amount of frames that is present in all micrographs, and I assume on datasets which are not
in the EER format, everything would work out fine from here on.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">However, since the amount of fractions of the original EERs is still different, I found that the program crashes with the error message like:<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">EERRenderer::renderFrames(frame_start = 967, frame_end = 987), NFrames = 918<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">EERRenderer::renderFrames(frame_start = 904, frame_end = 924), NFrames = 918<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">n: /Applic.HPC/Easybuild/zen3/2021a/build/RELION/4.0/foss-2021a-beta-543db0a-CUDA-11.3.1/relion-543db0a84a6735cc975be93fbafc0ffe683ebc57/src/renderEER.cpp, line 315<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">ERROR: <o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Invalid frame range was requested.<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">I found that when I limit the amount of fractions to sth like 20, relion would not crash (maybe because then the highest amount of fractions in one of the datasets that contributed to the first 20 frames is not lower
than the total amount of fractions in another subset). However, if 30 frames from one dataset are made from e.g. 750 fractions, while in the other subset the total 50 frames only were made from 690 fractions (because of higher exposure per second), the program
would crash with above error. <o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Is this a bug in relion? <o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">How can I overcome this? <o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Is the only way to first make movies with 50 frames from each subset and not work on the original EERs anymore? I processed fine to about 4 Angstroem without doing so, is it really necessary just for polishing to now
make 50 frame movies, or work on each subset separately? (As far as I understand relion, it does not have a GUI-function to generate 50 frame movies from EER data, but it has a command-line function to do so, am I right?<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Best wishes<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US">Björn<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US" style="mso-fareast-language:DE"><o:p> </o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
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