diff --git a/index.md b/index.md index 589a956..42587ab 100644 --- a/index.md +++ b/index.md @@ -24,6 +24,7 @@ views, images and tables for your colleagues to explore - [Locations and views](./tutorials/views_and_locations.md) - [Importing and exporting tables](./tutorials/importing_and_exporting_tables.md) - [Creating your own views](./tutorials/creating_your_own_views.md) +- [Volume Registration tutorial](./tutorials/volume_registration.md) - [Annotation tutorial](./tutorials/annotation_tutorial.md) - [Grid views & image region tables](./tutorials/image_grids_and_tables.md) - [More features](./tutorials/more_features.md) diff --git a/tutorials/tutorial_images/addCurrentImage.png b/tutorials/tutorial_images/addCurrentImage.png index 392f310..bd0c996 100644 Binary files a/tutorials/tutorial_images/addCurrentImage.png and b/tutorials/tutorial_images/addCurrentImage.png differ diff --git a/tutorials/tutorial_images/multiview.png b/tutorials/tutorial_images/multiview.png new file mode 100644 index 0000000..9a50055 Binary files /dev/null and b/tutorials/tutorial_images/multiview.png differ diff --git a/tutorials/volume_registration.md b/tutorials/volume_registration.md new file mode 100644 index 0000000..6394ecf --- /dev/null +++ b/tutorials/volume_registration.md @@ -0,0 +1,55 @@ +## Volume registration + +This tutorial will show how to generate a project from multimodal volume images and register them. + +### retrieve the raw data + + + - To download the Electron microscopy (EM) dataset, head to https://www.ebi.ac.uk/empiar/EMPIAR-11537/, make sure to de-select all data except #4 "Downscaled (20nm) aligned FIB SEM stack resliced to match the Airyscan dataset" and click the individual download button (choose HTTP). + + - The Fluorescence microscopy (FM) dataset is stored at https://www.ebi.ac.uk/biostudies/bioimages/studies/S-BSST1075. You can download the raw data file [directly](https://www.ebi.ac.uk/biostudies/files/S-BSST1075/EM04480_05_4G_Hoechst_GFP-TGN46_agglutinin_mitotracker.czi). + +### create the MoBIE project + +- open the LM data in Fiji using the BioFormats Importer plugin. (`Plugins > Bio-Formats > Bio-Formats Importer`) +- you should get a Hyperstack image with 4 channels. (MitoTracker, Agglutinin, GFP-TGN46, and Hoechst) +- adjust the lookup table for each channel to fit the fluorophore. (`Image > Lookup Tables`) +- Currently, the MoBIE project creator only supports single channels, so we have to split the fluorescence stack into separate channels using `Image > Hyperstacks > Make Subset...` and choose the channel number. Do this with each of the 4 channels active. +--- +- create a new MoBIE project: type "mobie" in the search bar or choose `Plugins > MoBIE > Create > Create New MoBIE project...` +![createMobieProject.png](tutorial_images/createMobieProject.png) + + name the project as you like and create it in a local directory. +- add a new dataset, name it as you like. Do not tick "2D". +![createDataset.png](tutorial_images/createDataset.png) +- select the first channel image in Fiji +- add a new source `current displayed image`, call it appropriately (channels are sorted inverse), make sure the Image type is "Image" and do not tick `make view exclusive`. +![addCurrentImage.png](tutorial_images/addCurrentImage.png) +- choose the `selection group name` as "fluorescence" or "FM" +![uiSelectionGroup.png](tutorial_images/uiSelectionGroup.png) + +- open the MoBIE project to see how the scaling and LUT are transferred + +--- +- continue the same way with the other channels, adding them to the same dataset and selection group. + +- open the other channel views using the MoBIE UI and explore the multi-channel volume +- right click into the multi-channel viewer and `Save current view` +- `Save as new view`, save to `projcect` +- Call the view something like "all channels" and make it part of the "FM" group +- close MoBIE +--- +- open the EM data in Fiji +- add the volume to MoBIE under the selection group "EM" (make new selection group) to the same dataset. The format conversion can take a couple of minutes. +- open the MoBIE project and view your multichannel FM and EM together +- switch off all fluorescence channels but the Hoechst by clicking on `S` +- change contrast and transparency settings for the relevant sources (`B`) + +![multiview.png](tutorial_images/multiview.png) + + +- right click the image "registration manual", select the EM image as we want to keep the multi-channles where they are, `Start manual transform` drag the image around. +- click `cancel manual Transform` to undo the translation and bring the image back to its original position. + +- use the mouse wheel to translate in `z`, make sure rotations are around the viewing axis and press the `z` key. Use the right and left arrow keys to rotate and the up and down to scale (dangerous) +- click `Accept manual Transform` to make it permanent and store the view into the project. This will only save the transformed EM source. Create a new selection group "Registration" \ No newline at end of file