-
Notifications
You must be signed in to change notification settings - Fork 26
New issue
Have a question about this project? Sign up for a free GitHub account to open an issue and contact its maintainers and the community.
By clicking “Sign up for GitHub”, you agree to our terms of service and privacy statement. We’ll occasionally send you account related emails.
Already on GitHub? Sign in to your account
register 3d images (X,Y,Z) and register from various objectives (x20 /x63) ? #133
Comments
Hi @Nal44, Best,
|
Hi Chandler, The first set is composed of several files 3D volumes (X,Y,Z), such as 5 individuals tif files (5 rounds per ROI) with 4 channels each (DAPI, beads, marker 1, marker 2). The cells shifted slightly between the rounds since it was acquired using multiple ROIS... For the other set, I have a 2d image (low mag ) and a higher one (x63 oil) on similar ROI. The idea is to register the 2d image (lower magnification ) onto the 3D image (higher magnification), then save as a OME.TIFF with 5 channels aligned (3d as 3 channels + 2d (dapi+marker) =5 channels total). Hence, it needs to be scaled then registered , so I can use a segmentation algorithm the get the masks on the 3D image and the registered 2d image and cross all markers for quantification . Let me know if this is clear or need more explanation, |
Hi @Nal44, For the second set of images. I think what you would want to do is make the 3D image your reference image ( Best, |
Hi Chandler, Thanks a lot, The way I did was to take the first Serie, segment the nucleus in 3d, then use this mask to manually align the other series to the first (translations/rotations) and applied the the same operations to the corresponding channels for each round (not very effective). These images are from collaborators, hence I will check first with them if I can share them (these are cells it should be ok I believe). For the second point I will try next week , I got delayed with too much work this week, |
Hi Chandler, Files are uploading now. Please let me know once received. thanks a lot for the help /support ! |
Hi @Nal44, Best, |
Hi Chandler, No probs, thank you for helping out ; D Please let me know, |
Hi @Nal44,
One can now align each volume to R1 by passing the above
Running the above produce the following overlap image, which I think looks pretty good: The
Running the above creates a volume with 16 planes, and 20 channels per plane. It also produces the ome-xml metadata which you can use to save the image using
It did take quite a bit of time to save this image (1.5h), but there may be ways to speed this up (different compression method, lower Please try out the above script and let me know how it goes. Best, |
Hi Chandler, Thanks a lot !! Great support and very helpful :D |
Hi,
I have a set of IF multiplex that I want to register but they are X.Y.Z (3D volume) ,
Using the code you sent me before (using the ch00 then match to all other channels with the same name) can I use the code or modify it to work with 3D images ?
Thanks a lot,
Antho
PS: also can I use valis to register images acquired from different objectives (x20 and x63) ? so scaling up/down before registering ?
Thanks
The text was updated successfully, but these errors were encountered: