PFC Projection mapping by Serial Two-photon tomography
- AAV-based fluorescent tracers are injected into the prefrontal cortices (PFC) of common marmosets.
- Fixed brain samples are embedded in agarose gel for sectioning and imaging.
Serial Two-photon Tomography (STPT) imaging
- Coordinated actions of vibratome and Z stage enables serial exposure of the blockface images.
- Two-photon imaging of the entire blockface by automated X-Y stage movement.
- Combined X/Y/Z imaging leads to distortion-free 3D data of the entire brain.
- Reconstruction of blockface image (2D) by tiling and stitching (typically 26×21 tiles) .
- Tracer signals and tissue autofluorescence are captured in different channels (green and red).
- 3D reconstruction can be simply done by using a stack of tiled and stitched images of the blockfaces.
- Algorithms are being developed to detect tracer signals and to registrate the sample brain to the reference.
[DataID: 2261. Creator Akiya Watakabe]