Virtual Histology with Photon Absorption Remote Sensing using a Cycle-Consistent Generative Adversarial Network with Weakly Registered Pairs

Modern histopathology relies on the microscopic examination of thin tissue sections stained with histochemical techniques, typically using brightfield or fluorescence microscopy. However, the staining of samples can permanently alter their chemistry and structure, meaning an individual tissue section must be prepared for each desired staining contrast. This not only consumes valuable tissue samples but also introduces delays in essential diagnostic timelines. In this work, virtual histochemical staining is developed using label-free photon absorption remote sensing (PARS) microscopy. We present a method that generates virtually stained histology images that are indistinguishable from the gold standard hematoxylin and eosin (H&E) staining. First, PARS label-free ultraviolet absorption images are captured directly within unstained tissue specimens. The radiative and non-radiative absorption images are then preprocessed, and virtually stained through the presented pathway. The preprocessing pipeline features a self-supervised Noise2Void denoising convolutional neural network (CNN) as well as a novel algorithm for pixel-level mechanical scanning error correction. These developments significantly enhance the recovery of sub-micron tissue structures, such as nucleoli location and chromatin distribution. Finally, we used a cycle-consistent generative adversarial network CycleGAN architecture to virtually stain the preprocessed PARS data. Virtual staining is applied to thin unstained sections of malignant human skin and breast tissue samples. Clinically relevant details are revealed, with comparable contrast and quality to gold standard H&E-stained images. This work represents a crucial step to deploying label-free microscopy as an alternative to standard histopathology techniques.