Objectives: Confocal laser endomicroscopy (CLE) is a non-invasive imaging modality of the gastrointestinal tract. Epithelial gaps in the small intestine of patients and rodents have been demonstrated using CLE. The goal of this study was to quantitatively validate the findings of epithelial gap density observed with CLE against confocal microscopy (CM) and light microscopy.
Methods: Two strains of mice (control 129 Sv/Ev and interleukin 10 knockout (IL-10(-/-))) underwent CLE of the terminal ileum. Adjacent ileal tissues were examined using CM and light microscopy. The total number of gaps and cells in the villi were manually counted from the three-dimensional reconstruction of cross-sectional CLE and CM images. The histology specimens were reviewed for epithelial gap and cell counts by a pathologist blinded to the study groups. The inter- and intra-observer variability for cell and gap counts were determined.
Results: For CLE, the gap densities (mean±s.d.) in the ileum for control and IL-10(-/-) mice were: 9.5±1.3 gaps per 1,000 cells and 20.6±2.1 gaps per 1,000 cells counted (P<0.001), respectively. For CM, the ileal gap densities were 7.3±1.3 gaps per 1,000 cells and 22.8±6.2 gaps per 1,000 cells (P=0.03), respectively. For light microscopy, the ileal gap densities were 29.2±5.9 gaps per 1,000 cells and 51.5±6.4 gaps per 1,000 cells for the two strains.
Conclusion: CLE can be used to quantitatively assess epithelial cells and gaps with accuracy comparable to CM and light microscopy. In a mouse model of inflammatory bowel disease, the epithelial gap density in the terminal ileum is significantly increased when examined using all three modalities.