Histopathology for Mohs Micrographic Surgery with Photoacoustic Remote Sensing Microscopy.

Mohs micrographic surgery (MMS) is a precise technique where layers of tissue are resected and examined with intraoperative histopathology to minimize the removal of normal tissue while completely excising the cancer. To achieve intraoperative pathology, the tissue is frozen, sectioned and stained over a 20- to 60-minute period, then analyzed by the MMS surgeon. Surgery is continued one layer at a time until no cancerous cells remain, meaning MMS can take several hours to complete. Ideally, it would be desirable to circumvent or augment frozen sectioning methods and directly visualize subcellular morphology on the unprocessed excised tissues. Employing photoacoustic remote sensing (PARS) microscopy, we present a non-contact label-free reflection-mode method of performing such visualizations in frozen sections of human skin. PARS leverages endogenous optical absorption contrast within cell nuclei to provide visualizations reminiscent of histochemical staining techniques. Here, we demonstrate the ability of PARS microscopy to provide large grossing scans (>1 cm^2, sufficient to visualize entire MMS sections) and regional scans with subcellular lateral resolution (400 +- 150 nm).