Abstract
BACKGROUND: The murine penis model has enriched our understanding of anomalous penile development. The morphologic characterization of the murine penis using conventional serial sectioning methods is labor intensive and prone to errors. AIM: To develop a novel application of micro-computerized tomography (micro-CT) with iodine staining for rapid, non-destructive morphologic study of murine penis structure. METHODS: Penises were dissected from 10 adult wild-type mice and imaged using micro-CT with iodine staining. Images were acquired at 5-μm spatial resolution on a Bruker SkyScan 1272 micro-CT system. After images were acquired, the specimens were washed of any remaining iodine and embedded in paraffin for conventional histologic examination. Histologic and micro-CT measurements for all specimens were made by 2 independent observers. OUTCOMES: Measurements of penile structures were made on virtual micro-CT sections and histologic slides. RESULTS: The Lin concordance correlation coefficient demonstrated almost perfect strength of agreement for interobserver variability for histologic section (0.9995, 95% CI = 0.9990-0.9997) and micro-CT section (0.9982, 95% CI = 0.9963-0.9991) measurements. Bland-Altman analysis for agreement between the 2 modalities of measurement demonstrated mean differences of -0.029, 0.022, and -0.068 mm for male urogenital mating protuberance, baculum, and penile glans length, respectively. There did not appear to be a bias for overestimation or underestimation of measured lengths and limits of agreement were narrow. CLINICAL TRANSLATION: The enhanced ability offered by micro-CT to phenotype the murine penis has the potential to improve translational studies examining the molecular pathways contributing to anomalous penile development. STRENGTHS AND LIMITATIONS: The present study describes the first reported use of micro-CT with iodine staining for imaging the murine penis. Producing repeated histologic sections of identical orientation was limited by inherent imperfections in mounting and tissue sectioning, but this was compensated for by using micro-CT reconstructions to identify matching virtual sections. CONCLUSION: This study demonstrates the successful use of micro-CT with iodine staining, which has the potential for submicron spatial resolution, as a non-destructive method of characterizing murine penile morphology. O'Neill M, Huang GO, Lamb DJ. Novel Application of Micro-Computerized Tomography for Morphologic Characterization of the Murine Penis. J Sex Med 2017;14:1533-1539.