Probe-based confocal laser endomicroscopy (pCLE) - a new imaging technique for in situ localization of spermatozoa

In azoospermic patients, spermatozoa are routinely obtained by testicular sperm extraction (TESE). However, success rates of this technique are moderate, because the site of excision of testicular tissue is determined arbitrarily. Therefore the aim of this study was to establish probe‐based laser endomicroscopy (pCLE) a noval biomedical imaging technique, which provides the opportunity of non‐invasive, real‐time visualisation of tissue at histological resolution. Using pCLE we clearly visualized longitudinal and horizontal views of the tubuli seminiferi contorti and localized vital spermatozoa. Obtained images and real‐time videos were subsequently compared with confocal laser scanning microscopy (CLSM) of spermatozoa and tissues, respectively. Comparative visualization of single native Confocal laser scanning microscopy (CLSM, left) and probe‐based laser endomicroscopy (pCLE, right) using Pro FlexTM UltraMini O after staining with acriflavine. Patients suffering of azoospermia – this means a lack of spermatozoa in the ejaculate – often have to undergo testicular sperm extraction (TESE). Up to now there is no imaging technique to precise the site of excision. This ex vivo study shows the feasibility of visualization of the testicular microarchitecture and the possibility to localize vital spermatozoa by probe‐based laser endomicroscopy (pCLE).