Label-Free Histology and Evaluation of Human Pancreatic Cancer with Coherent Nonlinear Optical Microscopy.

Surgeries achieving maximal tumor resection remain the major effective treatment of pancreatic cancer. Rapid and precise intraoperative diagnosis of pancreatic tissues is critical for optimum surgical outcomes but is challenging for the current staining-based histological methods. We demonstrated that label-free coherent nonlinear optical microscopy with combined stimulated Raman scattering (SRS) and second harmonic generation (SHG) could reveal key diagnostic features of both normal and cancerous human pancreatic tissues. Adjacent pairs of tissue sections from resection margins of 37 patients were imaged by SRS and hematoxylin and eosin staining for direct comparison, demonstrating high diagnostic concordance (Cohen's kappa, κ > 0.97) between them. Fresh unprocessed tissues showed well-preserved histoarchitectures including pancreatic ducts, islets, acini, and nerves. Moreover, the area ratios of collagen fibers were analyzed and found to correlate with the drainage pancreatic amylase level (odds ratio = 28.0, p = 0.0017). Our results indicated that SRS/SHG histology provides potential for rapid intraoperative diagnosis of pancreatic cancer as well as a predictive value of postoperative pancreatic fistula.

Differential characterization of lumbar spine associated tissue histology with nonlinear optical microscopy.

Percutaneous endoscopic lumbar discectomy (PELD) is the major effective treatment for lumbar disc herniation, and rapid histological identification of dissected tissue is critical to guide the discectomy. In this work, we revealed the histological features of different types of peridural tissues of the lumbar spine by label-free multi-modal nonlinear optical microscopy. Stimulated Raman scattering (SRS) was used to extract lipid and protein distributions, while second harmonic generation (SHG) and two-photon excited fluorescence (TPEF) signals were applied to image the collagen and elastin fibers at the same time. Our results demonstrated that the nonlinear optical features of the dura and adjacent soft tissues were significantly different, showing the potentials of our method for intraoperative differentiation of these critical tissues and improving the surgical outcome of PELD.