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    --- tags: WIP, NGFF --- Josh: would be good to know how much of this could be moved to either - https://github.com/ome/omero-cli-zarr - https://github.com/IDR/SubmissionWorkflow Comments from Petr for integrating: - 1. say whether or not the IMAGE_PATH can point to any old image or does it have to be an image inside the ManagedRepo of OMERO or otherwise “special” image - 2. say what is the output of the workflow and give a hint about how to follow-up (…and now open locally in napari) as applicable - (basically the need to explain the bigger framework, why are the omero steps there, and why and when can I omit/ignore them) # NGFF workflow, summarised A summarised version of https://github.com/IDR/SubmissionWorkflow/blob/master/zarr.md (private) that will be used to construct a workflow. Input parameters: - `IMAGE_ID`: OMERO Image ID - `IMAGE_PATH`: Path to raw data on NFS - `OUTPUT_DIR`: Output parent directory for Zarrs Temporary files passed between steps: - `SERIES_CSV`: CSV including image IDs - `ROIS_CSV`: CSV including ROI IDs Conda Environment: - bioformats2raw - omero-cli-zarr - omero-py - zarr Current workflow: ## 1. Obtain OMERO all image IDs associated with `${IMAGE_ID}`, save to ${SERIES_CSV}: omero hql --style=plain -q "select i.id, i.series from Image i where i.fileset.id = (select i.fileset.id from Image i where i.id = ${IMAGE_ID}) order by i.series asc" | cut -f2,3 -d, | tee ${SERIES_CSV} ## 2. Convert raw image data to Zarr bioformats2raw --file_type=zarr ${SERIES_CSV} ${IMAGE_PATH} ${OUTPUT_DIR} ## 3. Add OMERO metadata curl -o- http://idr.openmicroscopy.org/webclient/imgData/${IMAGE_ID}/ > ${OUTPUT_DIR}/${IMAGE_ID}.zarr/omero.json (Presumably need to loop through all image IDs in `${SERIES_CSV}`) ## 4. Merge external OMERO metadata into the Zarr `merge.py`: https://github.com/IDR/idr-zarr-tools/blob/master/merge.py ./merge.py {OUTPUT_DIR}/${IMAGE_ID}.zarr (Should we delete `${OUTPUT_DIR}/${IMAGE_ID}.zarr/omero.json`?) ## 5. Get mask IDs omero hql --style=plain "select distinct s.textValue, s.roi.id from Shape s where s.roi.image.id = ${IMAGE_ID}" --limit=-1 | tee ${ROIS_CSV} (Presumably need to loop through all image IDs in `${SERIES_CSV}`) ## 6. Add masks omero zarr masks Image:${IMAGE_ID} --mask-map=${ROIS_CSV} ---- Looks like steps 3-6 should be in a loop over the output of step 2. One question is whether to keep the loop in the same workflow, or to branch into a different workflow. # omero-cli-zarr export using Docker Based on https://github.com/ome/omero-cli-zarr/pull/29#issuecomment-713516616 and https://github.com/ome/omero-cli-zarr/pull/38#pullrequestreview-518083982 ``` $ ssh -A ome-zarr-dev1.openmicroscopy.org ``` Or use ``` ssh idr-ngff-1 ``` if you have: ``` Host idr-ngff-1 ProxyCommand ssh idr-pilot.openmicroscopy.org -W zarr1-dev:%p ``` Can run from your home directory... ``` $ mkdir idr0033_zarr_export_docker $ cd idr0033_zarr_export_docker $ vi Dockerfile ``` NB: Paste in this docker file, editing the omero-cli-zarr branch you want to use: ``` FROM centos:7 RUN yum install -y git python3 fontconfig RUN useradd -ms /bin/bash converter USER converter RUN python3 -m venv /tmp/venv && /tmp/venv/bin/pip install -U pip wheel && /tmp/venv/bin/pip install https://github.com/ome/zeroc-ice-py-centos7/releases/download/0.2.1/zeroc_ice-3.6.5-cp36-cp36m-linux_x86_64.whl RUN /tmp/venv/bin/pip install git+https://github.com/ome/omero-cli-zarr.git@refs/pull/38/merge # or RUN /tmp/venv/bin/pip install git+git://github.com/joshmoore/omero-cli-zarr.git@nested#egg=omero-cli-zarr COPY export.omero . ENV OMERO_USERDIR /tmp/omero ENTRYPOINT ["/tmp/venv/bin/omero", "load", "export.omero"] ``` Create a companion export.omero file ``` vi export.omero ``` And add export commands to connect to idr-testing... ``` login -C public@idr-testing.openmicroscopy.org -w public zarr --output /data export Plate:5966 ``` Then build with ``` $ sudo docker build --tag idr0033_plate5966 . ``` We want files to end up at e.g. (named after current or last-merged PR) https://minio-dev.openmicroscopy.org/idr/idr0033-rohban-pathways/41744_illum_corrected/pr35/5966.zarr Location is on /uod/idr/objectstore/minio/idr/ but this is now accessible from ome-zarr-dev1.openmicroscopy.org. Make sure target location exists... ``` $ cd /uod/idr/objectstore/minio/idr/idr0033-rohban-pathways/41744_illum_corrected $ mkdir pr_35 ``` Want to run with your own user ID, so you can modify output files ``` $ id wmoore # shows user ID - e.g. 5098 $ screen -xRR ``` NB: had some permissions problems (fixed) but the session created when the Dockerfile was built above then expired, so needed to run the container interactively and re-login to idr: ``` [wmoore@idr1-slot2 idr0002]$ sudo docker run -it --rm -u 5098 -e OMERO_USERDIR=/tmp/omero -v /uod/idr/objectstore/minio/idr/idr0033-rohban-pathways/41744_illum_corrected/pr_35:/data idr0033_plate5966 WARNING: Could not load OpenGL library. WARNING:vispy:Could not load OpenGL library. Server: [localhost:4064]idr.openmicroscopy.org Username: [root]public Password: Created session for public@idr.openmicroscopy.org:4064. Idle timeout: 10 min. Current group: Public Exporting to /data/5966.zarr 0.06% done, ETA: 05:20:46 ````` Should now be able to see files appearing on idr0-slot3 e.g. to see how many Wells have been exported: ``` $ cd /uod/idr/objectstore/minio/idr/idr0033-rohban-pathways/41744_illum_corrected/pr_35/5966.zarr/0 $ find . -maxdepth 2 -type d | grep [0-9] | wc 64 64 430 ``` # export with Z-downsampling Export and then downsample, working with Temp state of PR: https://github.com/ome/ome-zarr-py/pull/71#issuecomment-759404371 ``` ssh -A ome-zarr-dev1.openmicroscopy.org mkdir idr0077_zarr_z-downsample cd idr0077_zarr_z-downsample ``` Dockerfile: ``` FROM centos:7 RUN yum install -y git python3 fontconfig RUN useradd -ms /bin/bash converter USER converter RUN python3 -m venv /tmp/venv && /tmp/venv/bin/pip install -U pip wheel && /tmp/venv/bin/pip install https://github.com/ome/zeroc-ice-py-centos7/releases/download/0.2.1/zeroc_ice-3.6.5-cp36-cp36m-linux_x86_64.whl RUN /tmp/venv/bin/pip install git+https://github.com/will-moore/omero-cli-zarr.git@use_ome_zarr_to_build_pyramid RUN /tmp/venv/bin/pip install git+https://github.com/will-moore/ome-zarr-py.git@downsample_Z COPY export_zarr . ENV OMERO_USERDIR /tmp/omero ENTRYPOINT ["bash", "export_zarr"] ``` Create an `export_zarr` bash script: ``` /tmp/venv/bin/omero login -C public@idr-testing.openmicroscopy.org -w public /tmp/venv/bin/omero zarr --output /data export Image:9836831 /tmp/venv/bin/ome_zarr scale /data/9836831.zarr /data/9836831_zscale.zarr --downsample_z ``` Build and run ``` $ sudo docker build --tag idr0077_9836831_zscale_02 . $ sudo mkdir -p /uod/idr/objectstore/minio/idr/idr0077-valuchova-flowerlightsheet/zscale_02 $ sudo chown wmoore /uod/idr/objectstore/minio/idr/idr0077-valuchova-flowerlightsheet/zscale_02 $ id wmoore # 5098 $ screen -xRR $ sudo docker run -it --rm -u 5098 -e OMERO_USERDIR=/tmp/omero -v /uod/idr/objectstore/minio/idr/idr0077-valuchova-flowerlightsheet/zscale_02:/data idr0077_9836831_zscale_02 ``` # export via conda environment Use: ``` ssh -A ome-zarr-dev1.openmicroscopy.org ``` Or with this config: ``` # /.ssh/config Host pilot-zarr* ProxyCommand ssh idr-pilot.openmicroscopy.org -W %h.openmicroscopy.org:%p ForwardAgent yes ``` Use: ``` ssh pilot-zarr1-dev ``` First install Conda. Using first link in list at https://docs.conda.io/en/latest/miniconda.html#linux-installers ``` $ wget https://repo.anaconda.com/miniconda/Miniconda3-py39_4.10.3-Linux-x86_64.sh $ bash Miniconda3-py39_4.10.3-Linux-x86_64.sh ... Miniconda3 will now be installed into this location: /lifesci/groups/jrs/wmoore/miniconda3 ... ``` After re-opening a Terminal: ``` $ which conda ~/miniconda3/bin/conda ``` Using `environment.yml` at https://github.com/ome/NGFF-ELMI-2021-Workshop/blob/main/binder/environment.yml ... ``` wget https://raw.githubusercontent.com/ome/NGFF-ELMI-2021-Workshop/main/binder/environment.yml $ conda create -f environment.yml CondaValueError: The target prefix is the base prefix. Aborting. $ conda env create -f environment.yml CondaValueError: prefix already exists: /lifesci/groups/jrs/wmoore/miniconda3 $ conda env create --prefix ./envs -f environment.yml NB: could use -n myenv instead of --prefix? ... ERROR conda.core.link:_execute(699): An error occurred while installing package 'conda-forge::pyopengl-3.1.5-py_0'. Rolling back transaction: done LinkError: post-link script failed for package conda-forge::pyopengl-3.1.5-py_0 location of failed script: /lifesci/groups/jrs/wmoore/envs/bin/.pyopengl-post-link.sh ==> script messages <== <None> ==> script output <== stdout: Warning: Missing OpenGL driver, install with yum install mesa-libGL-devel or equivalent ... $ sudo yum install mesa-libGL-devel # try again... $ conda env create -f environment.yml -n omero_zarr_export ``` Seemed to work: ``` $ conda env list # conda environments: # /lifesci/groups/jrs/wmoore/envs base * /lifesci/groups/jrs/wmoore/miniconda3 omero_zarr_export /lifesci/groups/jrs/wmoore/miniconda3/envs/omero_zarr_export ``` Activate and install version of omero-cli-zarr we want to use ``` $ conda activate omero_zarr_export $ pip uninstall omero-cli-zarr $ pip install git+https://github.com/will-moore/omero-cli-zarr.git@export_v0.3_ome_ngff $ cd /uod/idr/objectstore/minio/idr/v0.3/idr0062-blin-nuclearsegmentation $ omero zarr export Image:6001240 ``` Available at: - 2D (YX): https://minio-dev.openmicroscopy.org/idr/v0.3/idr0094-ellinger-sarscov2/10503791.zarr - 3D (CXY): https://minio-dev.openmicroscopy.org/idr/v0.3/idr0077-valuchova-flowerlightsheet/9836842.zarr - 4D (TCYX) https://minio-dev.openmicroscopy.org/idr/v0.3/idr0002-heriche-condensation/179758.zarr - 4D (CZYX): https://minio-dev.openmicroscopy.org/idr/v0.3/idr0062-blin-nuclearsegmentation/6001240.zarr - 4D (TZYX): idr0099-jain-beetlelightsheet/12557113.zarr (not done) - 4D (TCYX): https://minio-dev.openmicroscopy.org/idr/v0.3/idr0077-valuchova-flowerlightsheet/9836849.zarr - 5D (TCZYX): https://minio-dev.openmicroscopy.org/idr/v0.3/idr0077-valuchova-flowerlightsheet/9836847.zarr (not done) Ended with: ``` OSError: [Errno 28] No space left on device: '/uod/idr/objectstore/minio/idr/v0.3/idr0099-jain-beetlelightsheet/12557113.zarr/0/0/54/0/0' ``` Seb: looks like we have used all the Inodes on the objectstore GPFS fileset at UoD so options are 1- clean up data on GPFS, 2- increase the number of inodes, 3- use EBI S3 rather than our minio Others: - https://minio-dev.openmicroscopy.org/idr/v0.4/2022-01-24/plates/idr0004 - https://minio-dev.openmicroscopy.org/idr/v0.4/2022-02-03/idr0062/6001240.zarr - https://minio-dev.openmicroscopy.org/idr/v0.4/2022-02-03/idr0101/13457227.zarr (13457537.zarr 13457539.zarr) - From bioformats2raw, includes OME/OME.xml for MIF https://minio-dev.openmicroscopy.org/idr/bf2raw/mdb/martin/sample_files.zarr/.zattrs is `{"bioformats2raw.layout" : 3}` # export vizarr & deploy on s3 Checkout https://github.com/hms-dbmi/vizarr/pull/43 $ cd vizarr $ npm run export # to /out directory # export via bioformats2raw Using pilot-zarr1-dev (ngff data directory: `/data`) or pilot-zarr2-dev (`/data/ngff`). Install bioformats2raw via conda: ``` conda install -c ome bioformats2raw ``` This is actually just for getting the dependencies installed. Get the actual bioformats2raw from this PR and just unzip it into your home directory: https://github.com/IDR/bioformats2raw/pull/1 Create a directory for the idr project and memo files (if it's not already there), and change into the idr directory. For example for idr0051: ``` mkdir idr0051 mkdir memo cd idr0051 ``` Find out where the pattern, screen or companion files are. For example: `/nfs/bioimage/drop/idr0051-fulton-tailbudlightsheet/patterns/` Then run the conversion (using the bioformat2raw from the PR!): ``` for i in `ls /nfs/bioimage/drop/idr0051-fulton-tailbudlightsheet/patterns/`; do echo $i; /home/<USER>/bioformats2raw-0.7.0-SNAPSHOT/bin/bioformats2raw --memo-directory ../memo /nfs/bioimage/drop/idr0051-fulton-tailbudlightsheet/patterns/$i ${i%.*}.ome.zarr; done ``` (`$i` is the pattern file, `${i%.*}.ome.zarr` strips the .pattern file extension and adds .ome.zarr; this should work for pattern, screen and also companion file extensions) # upload to uk1 s3 Install minio and configure... ``` ssh -A ome-zarr-dev1.openmicroscopy.org wget https://dl.min.io/client/mc/release/linux-amd64/mc chmod +x mc (base) [wmoore@ome-zarr-dev1 ~]$ ./mc config host add uk1s3 https://uk1s3.embassy.ebi.ac.uk Enter Access Key: X8GE11ZKP71A8529XFAE Enter Secret Key: Added `uk1s3` successfully. $ ./mc ls uk1s3/idr/zarr [2022-10-13 11:07:56 BST] 0B test-data/ [2022-10-13 11:07:56 BST] 0B v0.1/ [2022-10-13 11:07:56 BST] 0B v0.2/ [2022-10-13 11:07:56 BST] 0B v0.3/ [2022-10-13 11:07:56 BST] 0B v0.4/ $ cd /uod/idr/objectstore/minio/idr/v0.4/2022-02-03/ $ /lifesci/groups/jrs/wmoore/mc cp -r idr0101/ uk1s3/idr/zarr/v0.4/idr0101A/ ...idr0101/13457539.zarr/labels/0/2/0/0/0/0/0: 13.21 GiB / 13.21 GiB ━━━━━━━ ``` Also installed at on idrtesting-pilot to allow direct upload without conversion... ``` ssh -A idr-pilot.openmicroscopy.org ssh -A idrtesting-omeroreadwrite # install and config as above... $ cd /uod/idr/filesets/idr0138-lohoff-seqfish/20210917-Globus/ngff/TimEmbryos-102219/HybCycle_0/ $ /home/wmoore/mc cp -r MMStack_Pos0.ome.zarr/ uk1s3/idr/zarr/v0.4/idr0138A/TimEmbryos-102219/HybCycle_0/MMStack_Pos0.ome.zarr/ .../MMStack_Pos0.ome.zarr/labels/nuclei/4/0/0/0: 186.41 MiB / 186.41 MiB ━━━━━━━ ``` # Copy files from IDR ``` rsync -av -e "ssh -A idr-pilot.openmicroscopy.org ssh" "idrtesting-omeroreadwrite:/uod/idr/filesets/idr0070-kerwin-hdbr/20200414-Batch3-ftp/HDBR_BMP4_ISH/" . -P ``` # Test import to OMERO ``` $ ssh idr2-slot3.openmicroscopy.org $ cd /uod/idr-scratch/will-test/ $ aws s3 sync --no-sign-request --endpoint-url https://uk1s3.embassy.ebi.ac.uk s3://idr/zarr/v0.4/idr0050A/4995115.zarr 4995115.zarr $ conda activate omeropy $ echo $OMERODIR /lifesci/groups/jrs/wmoore/OMERO.server-5.6.5-ice36-b233 $ omero import --depth=100 4995115.zarr ``` # Update Jan 2023 See https://forum.image.sc/t/converting-other-idr-images-into-public-zarr/44025/12 Want to convert /uod/idr/filesets/idr0048-abdeladim-chroms/20181217-ftp/Astrop65_BDV/ (astroP65.h5 (73GB) and astroP65.xml) Seb: "`pilot-zarr1-dev` or `pilot-zarr2-dev` that has the advantage of being on EBI Embassy (so easier to upload to S3)" ``` ssh -A -o 'ProxyCommand ssh idr-pilot.openmicroscopy.org -W %h:%p' pilot-zarr1-dev ``` Installed conda and bioformats2raw as above in `/home/wmoore/miniconda`, and installed & configured `mc` client as above. ``` rsync -av -e "ssh -A idr-pilot.openmicroscopy.org ssh" "idrtesting-omeroreadwrite:/uod/idr/filesets/idr0048-abdeladim-chroms/20181217-ftp/Astrop65_BDV/" -P . ``` ``` $ bioformats2raw --version Version = 0.5.0 Bio-Formats version = 6.10.1 NGFF specification version = 0.4 $ ls -lh total 73G -rw-rw----. 1 wmoore idrnfs 73G Dec 17 2018 astroP65.h5 -rw-rw----. 1 wmoore idrnfs 2.3K Dec 17 2018 astroP65.xml $ bioformats2raw astroP65.xml 9846151.zarr OpenJDK 64-Bit Server VM warning: You have loaded library /tmp/opencv_openpnp3473542386768814439/nu/pattern/opencv/linux/x86_64/libopencv_java342.so which might have disabled stack guard. The VM will try to fix the stack guard now. It's highly recommended that you fix the library with 'execstack -c <libfile>', or link it with '-z noexecstack'. $ /home/wmoore/mc cp -r 9846151.zarr/ uk1s3/idr/zarr/v0.4/idr0048A/9846151.zarr/ ``` **conda install** ``` conda create -n omero_zarr_export -c ome python=3.9 zeroc-ice36-python omero-py conda activate omero_zarr_export # custom versions of ome-zarr-py and omero-cli-zarr pip install git+https://github.com/ome/ome-zarr-py.git@refs/pull/244/head pip install git+https://github.com/ome/omero-cli-zarr.git@refs/pull/134/head ``` # March 2023 Rsync local data to minio: ``` rsync -rvP --progress 13425213.zarr ome-zarr-dev1.openmicroscopy.org:/uod/idr/objectstore/minio/idr/idr0113-bottes-opcclones/ ``` # IDR NGFF conversion - make buckets - April 2023 ``` $ aws --endpoint-url https://uk1s3.embassy.ebi.ac.uk s3 mb s3://idr0010 ``` Seb: set the policy on the bucket as described in https://github.com/IDR/deployment/blob/master/docs/object-store.md#policy ? this will be mandatory for accessing it from the pilots (without keys) Will: also need to configure CORS as described there too. Update - April 2025 Minio hosted at `ome-dckr-ap1` which is at... ``` ssh 134.36.7.234 ls -alh /uod/idr/objectstore/minio/idr/Testing/sample_files.zarr/ ``` NB: Dom had to move the data there as when I put it at `/uod/idr/objectstore/minio/idr/ome-ngff-tools/sample_files.zarr/` it wasn't accessible via minio! NB: tried to create a new top-level bucket (sibling to idr) but couldn't fix CORS... ``` aws s3api put-bucket-cors --endpoint-url https://minio-dev.openmicroscopy.org --bucket ome-ngff-tools --cors-configuration file://cors.json ``` ... as I don't know aws credentials! Update: May 2025: `pilot-zarr1`, `pilot-zarr2` & `pilot-zarr3` will be replaced by `pilot-zarr`: see https://github.com/openmicroscopy/management_tools/pull/1798/files # August 2025 Want to install `mc` locally to upload to https://uk1s3.embassy.ebi.ac.uk... Install mc with: ``` $ conda create -n mc python=3.12 $ conda activate mc $ conda install "go-mc==2020.11.17" -c joshmoore ``` But then... ``` $ mc config host add uk1s3 https://uk1s3.embassy.ebi.ac.uk mc: Configuration written to `/Users/wmoore/.mc/config.json`. Please update your access credentials. mc: Successfully created `/Users/wmoore/.mc/share`. mc: Initialized share uploads `/Users/wmoore/.mc/share/uploads.json` file. mc: Initialized share downloads `/Users/wmoore/.mc/share/downloads.json` file. Enter Access Key: X8GE11ZKP71A8529XFAE Enter Secret Key: mc: <ERROR> Unable to initialize new alias from the provided credentials. Get https://uk1s3.embassy.ebi.ac.uk/probe-bucket-sign-deollovggy2v/?location=: x509: certificate signed by unknown authority. ```

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