Real-time Optical Diagnosis for surgical margin in low rectal cancer Using Multiphoton Microscopy

Jun Yan, MD, Shuangmu Zhuo, PhD, Gang Chen, Mingang Ying, MD

Department of Surgery, Fujian Provincial Tumor Hospital, Teaching Hospital of Fujian Medical University, Fuzhou, 350014, P.R.China; Institute of Laser and Optoelectronics Technology, Fujian Provincial Key Laboratory for Photonics Technology, Key Laborato

Background:
Multiphoton microscopy (MPM), based on the advancement of the field of non-linear optics and femtosecond lasers, has been shown to provide real-time detailed information on tissue architecture and cell morphology in live tissue. The purpose of this study was to evaluate the feasibility of using MPM to make real-time optical diagnosis for surgical margin in low rectal cancer.

Methods: Thirty fresh, unfixed, and unstained full-thickness surgical margins in low rectal cancer underwent MPM examination and then went through routine intra-operative pathological frozen procedure. MPM images were compared with golden standard hematoxylin-eosin (H-E) stained images.

Results: MPM images were obtained by two photon-excited fluorescence signals from tissue sample. Peak multiphoton autofluorescence intensity was detected in mucosa excited at 800 nm. In the normal area of surgical margin, MPM revealed regular tissue architecture and cell morphology, including a typical foveolar pattern with central, round crypt openings, and glands lined by epithelian cells and goblets cells. In the cancerous area of surgical margin, MPM demonstrated irregular tubular structures, reduced stroma, and cellular and nuclear pleomorphism. Cancer cells, characterized by irregular size and shape, enlarged nuclei, and increased nuclear-cytoplasmic ratio, were identified by MPM images, which were comparable to H-E stained images.

Conclusions: It is feasible to use MPM to make real-time optical diagnosis for surgical margin in low rectal cancer. With miniaturization and integration of colonoscopy or probe, MPM has the potential to provide real-time non-invasive “optical biopsy” for surgical margin in low rectal cancer in the near future.

Key words: Multiphoton microscopy; Real-time diagnosis; Surgical margin; Low rectal cancer; Pathology.

Session: Podium Presentation

Program Number: S043

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