Movie 2: 3D visualization of the adult human hypogastric nerve (HN)

A sample block of the adult human (male, 78 years) hypogastric nerve (HN) preserved by paraformaldehyde (PFA) fixation (unembalmed), followed by tissue clearing and light sheet fluorescence microscopy imaged using 561 nm autofluorescence (no antibody staining).Neural tracts and blood vessels can be clearly observed using autofluorescence without additional staining. The neural tracts run parallel to one another, and some contain embedded neuron clusters. A prominent vessel overlies these tracts and demonstrates a complex vascular branching pattern. Transverse optical slices using the Imaris Ortho Slicer tool shows these structures running parallel to one another for a region of the block. A putative paraganglion, which was later validated by immunostaining for tyrosine hydroxylase (TH) (not shown), has anatomical fascicular connections to the HN, is highly vascularised, and contains a small number of neurons embedded in it.Human tissue specimens were obtained from the organ donation program based at the Centre Hospitalier Universitaire de Nîmes (CHU de Nîmes), France and the Body Donor Program at the University of Melbourne, Australia. Ethical approval for the use of human tissue was granted from Human Research Ethics Committees at both institutions.