====== Pre-processing of dataset from Titan II ====== ==== 1. Recover the dataset from the microscope ==== Log into **cbi-gateway-01** to access the microscope and transfer the data to **gateway** or **storage** ssh @cbi-gateway-01 ls /mnt/titan2/offloaddata/TemScripting/EF-Falcon/ rsync -avz --progress /mnt/titan2/offloaddata/TemScripting/EF-Falcon/ /mnt/storage/teams/// Path to gateway as seen from cbi-gateway-01 : /mnt/zfspool/teams/ ==== 2. Convert the EER movies to TIF ==== Log into **cbi-compute-01** and transfer the scripts {{ ::eer2tif_parallel_loop.py |}} and {{ ::eer2tif_parallel.py |}} in your **home** ssh @cbi-compute-01 module load relion cd /mnt/storage/teams//// #go to the folder with the dataset module load python python ~/eer2tif_parallel.py Warning : conversion of eer to tiff file will show error this message : TIFFReadDirectory: Warning, Unknown field with tag 65002 (0xfdea) encountered. ==== 3. Begin processing with relion ==== Load and start relion (from cbi-compute-01 or the team GPU node (phantom-node39 for Lamour-Ruff)) module load relion relion& #start relion in the folder with movies === Job Relion/Import === Give the path to the *.tif , specify the pixel size, voltage and hit **RUN** You can **RESUME** the job when more movies are transferred and converted === Estimation the camera gain === Estimate the gain with at least 200 movies (redo if needed with a larger number) relion_estimate_gain --i Import/job001/movies.star --j 8 --max_frames 10000 --random true --o estimated_gain.mrc Explanation of options: * --j: number of thread * --max_frames: target number of frames to average (rounded to movies) * --random: randomize the order of input movies before taking subset === Job Relion/Motion correction === Specify the dose per frame, the number of patches 5x5m, provide the gain file, and set several MPI (number of tasks in parallel) and several (as a multiple of the number of frames in the movies) threads (number of cpu per task)