Colonic crypt morphological patterns have shown a close correlation with histopathological diagnosis. Imaging technologies such as high-magnification chromoendoscopy and endoscopic optical coherence tomography (OCT) are capable of visualizing crypt morphology in vivo. We have imaged colonic tissue in vitro to simulate high-magnification chromoendoscopy and endoscopic OCT and demonstrate quantification of morphological features of colonic crypts using automated image analysis. 2-D microscopic images with methylene blue staining and correlated 3-D OCT volumes were segmented using marker-based watershed segmentation. 2-D and 3-D crypt morphological features were quantified. The accuracy of segmentation was validated, and measured features are in agreement with known crypt morphology. This work can enable studies to determine the clinical utility of high-magnification chromoendoscopy and endoscopic OCT, as well as studies to evaluate crypt morphology as a biomarker for colonic disease progression.