Figure 1. Development of the siGOLD method.
(A) Schematic flowchart of the siGOLD labeling approach from high-pressure freezing and freeze substitution (HPF-FS) to tracing and 3D reconstruction. Ni, nickel grid, Cu, copper grid. (B) SEM micrograph of a 72 hpf Platynereis larva. (C) Schematic of the HT9-5 sample showing the position of the ventral nerve cord (VNC), ventral view. Colored lines indicate where cross-sections through the VNC were taken, near the base of the circumesophageal connectives at the level of the first commissure. Layer number(s) followed by neuropeptide ID are indicated for each colored line. Dashed line indicates the gap (approximately 10 missing sections) between the first and second series of sections. Scale bar: (B) 50 μm.