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

Abstract

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 labelfree 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.

Seeing is Believing: Developing Multimodal Metabolic Insights at the Molecular Level

“…SRS imaging facilitates rapid, label-free visualization of tissue, characterized by its unique capacity for chemical contrast, enabling the discernment of alterations such as protein-to-lipid ratio within tumor-infiltrated tissue and variations in cellularity . It can also be used for tissue histology due to the chemical contrast of intrinsic metabolites . MALDI-MSI offers comprehensive molecular insights across a wide mass spectrum, spanning small molecules to large proteins, , by generating detailed 2D ion images of specific compounds that may have otherwise not been detected in the fingerprint region of the SRS imaging.…”

“…114 It can also be used for tissue histology due to the chemical contrast of intrinsic metabolites. 173 MALDI-MSI offers comprehensive molecular insights across a wide mass spectrum, spanning small molecules to large proteins, 174,175 by generating detailed 2D ion images of specific compounds that may have otherwise not been detected in the fingerprint region of the SRS imaging. The capacity to correlate chemotypes in tissue regions through MSI renders it valuable for investigating disease progression, discovering biomarkers, and enhancing diagnostic precision.…”

Histological diagnosis of unprocessed breast core-needle biopsy via stimulated Raman scattering microscopy and multi-instance learning

“…It has demonstrated remarkable potentials in rapid histology for various types of human tissues and diseases, including brain tumor, laryngeal carcinoma, gastrointestinal tumor, pancreatic tumor and neurodegenerative disease, etc. [25,[29][30][31][32][33][34][35]. The acquired chemical species commonly include lipid and protein imaged by SRS, and collagen fibers taken by second harmonic generation (SHG) [31,36].…”

“…[25,[29][30][31][32][33][34][35]. The acquired chemical species commonly include lipid and protein imaged by SRS, and collagen fibers taken by second harmonic generation (SHG) [31,36]. On the other hand, various machine-learning models have been applied on SRS images for the analysis, classification and segmentation of tumor histopathology [29,30,[37][38][39].…”

Current Trends of Raman Spectroscopy in Clinic Settings: Opportunities and Challenges

“…Furthermore, they explored the potential of machine and deep learning for automating the identification and analysis of crucial histological features. [ 30 ] The significance of these studies extends beyond technological advancements in the domain of medical image diagnostics. They also provide clinicians with expedited and more precise diagnostic tools, which are particularly pivotal in the early detection and treatment of ailments such as cancer.…”