Label-free high-resolution volume imaging of cleared testes.

Accurate testicular tissue characterization is critical for diagnosing reproductive disorders and malignancies. We present a rapid, label-free 3D imaging technique that leverages endogenous autofluorescence and a fast, non-toxic optical clearing protocol to reveal the full 3D architecture of thick mouse testis samples, without sectioning. This approach clearly distinguishes structures like epithelial tubules, interstitial tissue, and key cell types (germ, Sertoli, Leydig) with high precision. Autofluorescence serves as a powerful structural counterstain, enhancing multiplexed analysis alongside immunofluorescent or transgenic markers. Compatible with high-resolution techniques like image-scanning confocal microscopy (AiryScan), our approach captures fine intracellular details while avoiding the artefacts of traditional histology. Accessible, affordable, and equally usable on standard confocal microscopes, this method democratizes advanced testis imaging and accelerates progress in reproductive research.