Novel multiphoton microscopy probe and feasibility study for colon cancer: Probe on trace elements–related antioxidant.

Abstract

463 Background: Multiphoton microscopy (MPM) has gained increasing popularity during the past few years because of the distinct advantages over single-photon microscopy, which includes increased penetration depth and low out-of-focus photodamage. However, there had been no report on multiphoton probes for malignancy. Previous studies reported that Zn and Cu ions which are co-factor of antioxidants such as superoxide dismutase (SOD), altered in malignancy tissue. The aim of this study was to evaluate application of the multiphoton (MP) probe of Zn and Cu for colon cancer tissues. Methods: We monitored the Zn/Cu ions in the colon cancer cell lines with multiphoton probe Microscopy. The multiphoton probe AZn2+ (C38H38N6O3) and ACu+ (C34H47N3O3S4), we developed and obtained the patent on it, were stained in live cancer cell lines. The tissues of colon cancer, adenoma and normal mucosa were obtained by biopsy during colonoscopy. Then the tissues were stained with 20μM of the MP probes for Zn/Cu ions. The distribution and concentration of zinc and copper ions were monitored by MPM by following the change in MPEF along the depth of tissue. Results: The majority of Zn2+ and Cu+ ions distributed in cytosole. The average multi-photon excited fluorescence (MPEF) intensities due to Cu, Zn, and Cu/Zn ratio were remarkably different between the normal mucosa and adenoma/adenocarcinoma tissues. The Zn2+ content was significantly lower and the Cu2+ content was significantly higher to result in a much lower the Zn/Cu ratio in adenoma/adenocarcinoma than in normal mucosa tissues. Conclusions: We have obtained multiphoton microscopy images of normal and cancer cell lines as well as mucosa and adenoma/adenocarcinoma tissues labeled with newly developed MP probes AZn1 and ACu1. No significant financial relationships to disclose.