Non-obstructive azoospermia (NOA), a severe male infertility condition with impaired or absent sperm production, is treated by microsurgical testicular sperm extraction (micro-TESE), whose success depends on identifying seminiferous tubules with active spermatogenesis. To address this challenge, we demonstrate that mid-infrared photothermal (MIP) microscopy can provide label-free molecular signatures to distinguish different NOA subtypes in patient tissues. We applied MIP microscopy and MIP-guided IR spectroscopy to testicular tissues from obstructive azoospermia (normal spermatogenesis) and idiopathic NOA (abnormal spermatogenesis) patients. Tissue classification was performed using a Singular Value Decomposition–Random Forest (SVD-RF) pipeline. MIP imaging revealed distinct lipid distribution and reduced lipid content in NOA tissues compared to normal spermatogenic tissues. Using SVD to extract spectroscopic features and RF for classification, we achieved 94.03% accuracy in distinguishing testicular tissues as normal spermatogenesis or three pathological subtypes of idiopathic NOA. These findings demonstrate MIP microscopy as an effective tool for characterizing the spermatogenic potential of seminiferous tubules based on their molecular composition, potentially facilitating improved sperm retrieval strategies.
Wu et al. (Fri,) studied this question.