Photoacoustic remote sensing 3D H&E histology with fluorescence validation

Hematoxylin and Eosin (H and E) staining is the gold standard for the majority of histopathological diagnostics but requires lengthy processing times not suitable for point-of-care diagnosis. Here we demonstrate a 266-nm excitation Ultraviolet Photoacoustic Remote Sensing (UV-PARS) and Scattering Microscopy system capable of virtual H and E 3D imaging of tissues in conjunction with with confocal fluorescence microscopy (CFM) for validation in thick tissues. We demonstrate the capabilities of this dual-contrast system for en-face planar and volumetric imaging of human tissue samples exhibiting high concordance with the gold standard of H and E staining procedures as well as confocal fluorescence microscopy. To our knowledge, this is the first near real-time microscopy approach capable of volumetric imaging unstained thick tissues with virtual H and E contrast.