The fly brain is a highly stereotyped 3D structure. A number of groups have now published 3D confocal image datasets where all images have been registered to a specific template brain. For almost all purposes the fly brain should be considered symmetric about its mid-sagittal plane i.e. the plane perpendicular to the medio-lateral axis - typically the X axis in image data. However, while the platonic fly brain may be symmetric, individual fly brains that have been fixed, stained and imaged are often
- significantly asymmetric
- not placed exactly in the centre of the image with respect to their mid-sagittal plane
While it is trivial to make template brains that are centred and possible to make symmetric template brains if one wishes, this has rarely been done in practice. It is therefore necessary to do more than simply mirror along the medio-lateral axis if one wishes to map neurons/structures in the left brain hemisphere onto the right hemisphere.
Each mirror registration in this repository maps one common template brain onto a medio-laterally flipped version of itself. It must therefore be combined with a preliminary step that flips the starting (image or point) data about the mid plane of the template image (typically YZ plane). The non-rigid registration therefore serves to fix the small displacements that are left after this simple flip.
The naming convention for the preliminary rigid registration about the mid plane of the template image is TARGET_imgflip.list where TARGET is a short name for the final template brain. The subsequent non-rigid registration to fix any residual displacements due to asymmetries in the brain are called TARGET_mirror.list. Thus to map image data from the right to left (or vice versa) in the JFRC2 template space (see virtual fly brain for details) one would want to use JFRC2_imgflip.list followed by JFRC2_mirror.list.
Of course it is also possible to avoid the use of the initial imgflip registrations when working with image data by using your favourite tool to flip the input data yourself along the correct axis. This could be as simple as Flip Horizontally in ImageJ. However you must ensure that the image's bounding box is not altered by this procedure – for ImageJ this means that images with non-zero origins cannot be handled.
All registrations were constructed with the aid of the open source Computational Morphometry Toolkit (CMTK). In each case the final output contains a non-rigid (warping) intensity-based registration between two template brains (CMTK command warp). In certain cases the initial affine that was used as a starting step was computed using a surface or landmarks-based registration rather than CMTK's intensity based registration (registration). You will need to install a recent (>=2.2) version of CMTK to use these registrations.
First download (or git clone) this repository
We recommend that you convert all images to the NRRD format. Fiji/ImageJ can read and write this format.
reformatx -o fully_mirrored_image.nrrd --floating simple_flipped_image.nrrd template_brain.nrrd target_mirror.list
Note that reformatx uses the template_brain.nrrd to determine the size and spatial location of the block of image data to produce. If you do not have the template brain, you can still produce output by manually defining the region that you want using reformatx's --target-grid option.
--target-grid <string>
Define target grid for reformating as Nx,Ny,Nz:dX,dY,dZ[:Ox,Oy,Oz] (dims:pixel:offset)
For details
reformatx --help
White space separated 3d coordinates can be converted (in either direction) using
streamxform -- --inverse target_source.list < start_coords.txt > mirror_coords.txt
streamxform target_source.list < target_coords.txt > sample_coords.txt
Note the use of the --inverse switch when mapping coordinates from source space to target space. This may be the opposite of what you expect but originates from the fact that in order to map a block of image data from source space to target space, the transformation that must actually be defined is the one that maps a regular grid of points in the target space back to sample locations in the source space. Note also the use of an extra -- to separate the positional arguments from the other options when using --inverse to specify that a given registration should be inverted.
For details
streamxform --help
Map a prominent point in the PLP from left to right
cd /path/to/MirrorRegistrations
echo 430 142.5 100 | streamxform -- --inverse JFRC2_imgflip.list --inverse JFRC2_mirror.list
# 206.67601 146.293051 105.666597
Now we can see how good our mirroring is
echo 206.67601 146.293051 105.666597 | streamxform -- --inverse JFRC2_imgflip.list --inverse JFRC2_mirror.list
# 429.836709 142.154807 99.4046885
i.e. close but not exactly the same point - due to the fact that the registration cannot perfectly match the two halves of the brain.
We can test the numerical stability of CMTK's inversion of the warping transformation by doing
echo 206.67601 146.293051 105.666597 | streamxform -- JFRC2_mirror.list JFRC2_imgflip.list
# 430 142.5 99.9999995
i.e. errors due to numerical instabilities in the inversion are minimal in this case
These registrations should be very accurate because the non-rigid registration is operating on data from exactly the same specimen. This can be tested by calculating the round trip error
P->P'->P''
By way of example for the JFRC2 template we find a mean displacement of 0.7 microns. Of course the displacements are not uniform and structurally variable parts of the brain (e.g. antennal lobes) will likely show larger displacements.
In comparison across brain registration errors might be order 3-4 microns.
A simple Bash script for the automatic production of a mirror registration, given a template image, is also provided in autoflip.sh. This requires a small amount of configuration to set the relative locations of reference brain images, the path to the munger program (part of CMTK), and the path to the bin directory containing CMTK commands. This script uses Fiji to mirror a given template image and then uses CMTK to perform an affine and non-rigid (warping) registration. As an example, to produce a registration for a JFRC.nrrd image saved in the reference brain directory, use ./autoflip.sh JFRC.
If you need help, please visit https://groups.google.com/forum/#!forum/nat-user.