Visualization of the human enteric nervous system by probe confocal laser endomicroscopy: a first real-time observation of Hirschsprung’s disease and allied disorders

Abstract

Background Our group previously proved that the human enteric nervous system can be visualized with confocal laser endomicroscopy after topical application of cresyl violet using surgically resected intestine specimens. The present report documents the first in vivo visualization of the human enteric nervous system with confocal laser endomicroscopy using local cresyl violet staining. The aim of this study was to evaluate the technical feasibility and clinical efficiency of confocal laser endomicroscopy in patients with Hirschsprung’s disease and allied disorders in vivo. Methods Confocal laser endomicroscopy was performed in vivo in two patients to confirm the presence of the enteric nervous system during surgery in patients with Hirschsprung’s disease and allied disorders. Cresyl violet was gently injected from the serosal side into the muscular layer of the intestine, and scanning was performed within 30 min. Then, the scanned intestines were resected, and the visualized area of the specimens was pathologically evaluated. Results The ganglion cell nuclei and the enteric nervous system network were clearly visualized intraoperatively in both cases. The morphological findings were similar to the pathological findings of the enteric nervous system in both cases although the period of visibility was brief. Conclusion This study demonstrated the first, real-time observation of the enteric nervous system in humans using confocal laser endomicroscopy and suggest the potential to identify the enteric nervous system intra-operatively during surgery for Hirschsprung’s disease and allied disorders.

Visualizing enteric nervous system activity through dye-free dynamic full-field optical coherence tomography

“…However, these technical approaches can be performed only ex vivo as they require tissue clearing and immunohistochemical staining. In vivo visualization of the human ENS was achieved using probe confocal endomicroscopy (pCLE) but as the resolution and penetration depth were too low, endoscopic submucosal dissection 6 or cresyl violet staining 6 was necessary to obtain a clear image of the ENS 7 . In this context, novel technologies providing high spatial resolution and dye-free dynamic imaging of the ENS is of great interest.…”