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.

• 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]