Accurate and fast histological assessment of freshly removed tissue specimens is of critical importance in clinical procedures such as diagnostic biopsy and surgical tumor resection. The current approach of frozen section analysis (FSA) for in-procedure histopathology is both time-consuming and damages the tissue sections used to the point of being unable to re-analyze them using traditional pathological staining methods . An alternative to FSA is the application of fluorescent stains that are able to mimic the pathological features of traditional hematoxylin and eosin (H&E) staining, followed by rapid non-destructive ex vivo microscopy, which conserves the tissue for future pathological and molecular analysis. Fluorescent staining is quick, on the order of seconds, and when paired with an appropriate microscopy system, can allow virtual H&E images of fresh tissue surfaces within minutes of removal from the patient. In this work we demonstrate a novel dual-stain fluorescent analog to H&E on prostate tissue sections and fresh animal tissues using confocal microscopy.
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