Nonlinear Microscopy System and Protocol for Rapid Evaluation of Freshly Excised Human Tissue

Abstract

Histopathology determines disease type/condition by microscopic evaluation of biopsy and surgically excised tissue, playing a critical role in medicine. The clinical protocol for histology requires physical sectioning of tissue, either with intense chemical processing and paraffin embedding or freezing of tissue, limiting the rapid evaluation applications. Nonlinear microscopy (NLM) is an emerging optical sectioning microscopy technology that enables histological visualization of fresh and intact human tissue without requiring physical sectioning.

In this thesis, we developed high-throughput, real-time NLM imaging system and protocol for intraoperative NLM evaluation of surgically excised tissue. We demonstrated the versatile imaging capability of NLM with comparative studies between NLM and other optical sectioning microscopy techniques. Interventional randomized clinical trials are designed and conducted to demonstrate the feasibility of intraoperative NLM evaluation (breast lumpectomy and radical prostatectomy). A pilot study demonstrates the feasibility of NLM imaging of bone. The studies in this thesis were performed in close collaboration with the Beth Israel Deaconess Medical Center. This thesis aims to develop NLM system and protocol for rapid evaluation of fresh human tissue and to design and perform clinical trials for validation of the efficacy of intraoperative NLM evaluation. The results suggest the practical potential of NLM as a modality to improve/transform clinical/surgical procedures.