Analytical evaluation of intact human tissues using mid-frequency scanning acoustic microscopy at the mesoscopic scale

Conventional histological examination requires irreversible sectioning, consuming tissue and precluding three-dimensional analysis. This limitation is particularly problematic for archival specimens and forensic investigations. Scanning acoustic microscopy (SAM) enables non-destructive tissue visualization through acoustic impedance contrast without sectioning or staining, offering a complementary approach to conventional histology. We investigated mid-frequency (50 MHz) SAM imaging of intact, formalin-fixed human tissue blocks representing diverse anatomical structures, including skull table, pineal gland, cerebellum, and scalp. SAM successfully delineated histologically meaningful microarchitectural features, including trilaminar skull organization with diploic channels; pineal calcifications (corpora arenacea); cerebellar cortical lamination with identifiable Purkinje cells; and stratified scalp architecture. Multifocal image integration and stereoscopic reconstruction enabled extended depth-of-field imaging and three-dimensional visualization of intact specimens. Corresponding hematoxylin–eosin and Masson’s trichrome histology confirmed close structural agreement with SAM-derived contrasts. This non-destructive imaging approach preserves tissue integrity while generating digital datasets suitable for volumetric reconstruction and downstream computational analysis, supporting applications in anatomical education, tissue banking, pathology training, and biomedical research.