Abstract
Background
The distribution of ganglion cells in the transition zone of Hirschsprung Disease
(HD) colons is extremely variable. Determining the resection margin based on intraoperative
biopsies may be imprecise. Multiphoton microscopy (MPM) is a novel imaging technology
with the ability to visualize tissues in real time. In this study, we evaluate the
potential of MPM to quantify ganglion cells in a murine model of HD.
Methods
After IACUC approval, formalin-fixed colons from 7 wild type (WT) and 6 Endothelin
Receptor B gene (EdnrB) homozygous knockout (KO) mice with distal colonic aganglionosis
were assessed by MPM for the presence of myenteric ganglion cells. MPM images were
captured starting from the anus progressing proximally at 5 mm intervals. Hematoxylin and eosin (H&E) stained biopsies of the imaged were correlated
with MPM findings.
Results
WT specimens showed normal myenteric plexus ganglia throughout the examined colon.
In contrast, distal colons of EdnrB KO animals were devoid of ganglia up to 10 mm from the anus. Ganglion cells were visible starting at 20–30 mm proximal to the anus. The density of ganglion cells seen by MPM and histology correlated
well.
Conclusions
MPM can clearly identify the myenteric plexus ganglia in both WT and KO mouse colons.
Comparison with the H&E-stained sections showed reproducible correlation. MPM-based
real-time imaging of the myenteric plexus may become a useful intraoperative decision-making
tool in the future.
Key words
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Article info
Publication history
Accepted:
March 8,
2013
Received:
February 23,
2013
Identification
Copyright
© 2013 Elsevier Inc. Published by Elsevier Inc. All rights reserved.
