DiI Staining of Fine Branches of Bonghan Ducts on Surface of Rat Abdominal Organs

Novel thread-like structures and corpuscles, designated Bonghan ducts (BHDs) and corpuscles (BHCs), are known to form a system of networked channels. Here, we tested the effectiveness of a fluorescent carbocyanine dye, DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate), in staining BHDs and BHCs. DiI solution was infused into a BHC on the surface of a rat abdominal organ at a steady rate and the resulting labeling of neighboring BHCs connected via BHDs was examined, as identified by the red fluorescence of DiI. BHDs diameters tapered away from BHCs and formed tree-like branches with fine arborizations embedded in the membranous tissues at their terminal parts. In the proximal parts, DiI fluorescence appeared as continuous lines within BHDs, but a large portion of BHDs remained unstained. In the distal parts of BHDs, discontinuous elongated DiI microparticles were identified along the sinuses within BHDs. The results showed that inner spaces within the BHDs allowed DiI to flow and that BHDs have tree-like branches and terminal arborizations. In conclusion, DiI can be used in visualizing BHDs fine structures.