Pathologists evaluate histology sectioned perpendicular to the tissue surface, or in vertical cross-sections. This orientation (X-Z plane) enables evaluation of mucosal differentiation in the basilar-to-luminal direction. Current endomicroscopes use a conventional (single axis) optical design. Imaging is limited to horizontal cross-sections (X-Y plane) where the microanatomy is frequently similar across the field of view (FOV). In the dual axes configuration, light is delivered and collected off-axis, and images can be detected over a much larger range of intensities. Molecular images collected using fluorescence can improve specificity for disease detection and reveal functional properties about tissue.Proper interpretation of these images requires correlation with the microanatomy. We aimed to demonstrate the simultaneous collection of 2 fluorescence images in vivo in vertical cross-sections using a dual axes confocal endomicro-scope. An overlay of molecular and anatomic images from normal and dysplastic mouse colonic mucosa are displayed in real time.
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