In vivovisualization of collagen fiber produced by cultured osteoblasts using sensitive second-harmonic-generation microscopy equipped with a 10-fs mode-locked Ti:sapphire laser

Abstract

Second-harmonic-generation (SHG) microscopy is a new tool for observing the collagen fiber in tissue in vivo. Conventional SHG microscopy equipped with 100-fs pulse laser is insufficient to visualize low-order-structured, immature collagen with high contrast (for example, collagen fiber produced by cultured cell) because of low nonlinear susceptibility χ (2) . To enhance the image contrast while avoiding the sample damage, one should increase a peak power of the laser light while maintaining the same average power as the 100-fs laser. In this paper, we constructed sensitive SHG microscopy equipped with a 10-fs Ti:Sapphire laser and succeeded to visualize collagen fibers produced by the cultured osteoblasts in vivo.