Abstract: Histopathology is the gold standard for disease diagnosis. The use of digital histology on fresh samples can reduce processing time and potential image artifacts, as label-free samples do not need to be fixed nor stained. This fact allows for a faster diagnosis, increasing the speed of the process and the impact on patient prognosis. This work proposes, implements, and validates a novel digital diagnosis procedure of fresh label-free histological samples. The procedure is based on advanced phase-imaging microscopy parameters and artificial intelligence. Fresh human histological samples of healthy and tumoral liver, kidney, ganglion, testicle and brain were collected and imaged with phase-imaging microscopy. Advanced phase parameters were calculated from the images. The statistical significance of each parameter for each tissue type was evaluated at different magnifications of 10×, 20× and 40×. Several classification algorithms based on artificial intelligence were applied and evaluated. Artificial Neural Network and Decision Tree approaches provided the best general sensibility and specificity results, with values over 90% for the majority of biological tissues at some magnifications. These results show the potential to provide a label-free automatic significant diagnosis of fresh histological samples with advanced parameters of phase-imaging microscopy. This approach can complement the present clinical procedures.

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