Fast and label-free 3D virtual H&E histology via active phase modulation-assisted dynamic full-field OCT.

Pathological features are the gold standard for tumor diagnosis, guiding treatment and prognosis. However, the standard histopathological process is labor-intensive and time-consuming. Although frozen sections offer a quicker alternative, they often induce severe artifacts and suffer from lower accuracy. Dynamic full-field optical coherence tomography (D-FFOCT) is an innovative optical imaging technique that provides rapid histological information by utilizing subcellular dynamics as an intrinsic source of contrast. Despite its advantages, D-FFOCT images of adjacent tissues frequently exhibit abrupt shifts in hue and brightness, which is confusing for pathologists and diminishes its interpretability and reliability. Here, we present an active phase modulation-assisted D-FFOCT (APMD-FFOCT) to improve the imaging stability and achieve continuity and consistency in image stitching, which also enhances the image contrast of tissues with low metabolic dynamics. This enables us to further employ an unsupervised generative adversarial network to convert APMD-FFOCT images into virtual hematoxylin and eosin (H&E) stained images for the first time. Three-dimensional (3D) virtual H&E-stained images can be obtained at a scanning rate of 1 frame per second. Furthermore, we also demonstrate that this novel technique has been successfully applied in cancer diagnosis for the human central nervous system and breast, which proves that this new method will play a new unique and important role in intraoperative histology.