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

Pathological features are the gold standard for tumor diagnosis, guiding treatment and prognosis. However, standard histopathological process is labor-intensive and time-consuming, while frozen sections have lower accuracy. Dynamic full-field optical coherence tomography (D-FFOCT) offers rapid histologic information by measuring the subcellular dynamics of fresh, unprocessed tissues. However, D-FFOCT images suffer from abrupt shifts in hue and brightness, which is confusing for pathologists and diminish their interpretability and reliability. Here, we present active phase modulation-assisted D-FFOCT (APMD-FFOCT) to improve the imaging stability and enhance the contrast of static tissues. This enables us to further employ an unsupervised deep learning 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 have been obtained at a scanning rate of 1 frame per second, as demonstrated in cancer diagnosis for human central nervous system and breast. The results prove that this new method will play a unique and important role in intraoperative histology.