Development of molecular imaging microscope for biological specimen and its application for tissue examination

生物标本分子成像显微镜的研制及其在组织检查中的应用

基本信息

批准号：
15300161
负责人：
ARAKI Tsutomu
金额：
$ 10.69万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15300161/
关键词：
bio-SHG nonlinear optical effect ultra-short duration pulse laser collagen molecule tissue diagnosis microscopic image skin

项目摘要

Aim of this research project is to develop an effective method that can sense objective biomolecles without staining and to apply this method to identification of the situation of the molecles. For ths purpose, we have focused on non-linear optical effect in the tissue that is induced by the irradiation with high-intensity very short duration pulsed laser light such as a femtosecond laser. In the actual research, we have developed three microscopes ; fluorescence lifetime imaging microscope in nanosecond range, a collagen sensitive microscope using second harmonic generation light (SCG light) and a coherent anti Stokes coherent Roman scattering (CARS) microscope.The most successful one is the SHG microscope. Based on the reflection-type microscope with a probe light spot of 10 micrometers in diameter and the polarization measurement of SHG light induced by collagen molecules, we have determined distribution of collagen fiber orientation in various human tissues. The resulting SHG data in human skin imply that the reticular dermis posses a tangled structure of collagen fibers which is consistent with the result of the anatomical examination of the skin. We have improved the SHG microscope to obtain 3D dissection image using the con-focal optics and the 3D sample scanning stage. With this microscope, tomograph of the collage fiber orientation in the human skin and bone have been measured.Performance of the fluorescence lifetime imaging microscope was demonstrated by the measurement of the autofluorescence in liposome. Also we have measured the distribution of hyaluronic acid around an erythrocyte with CARS microscope.

该研究项目的目的是开发一种有效的方法，该方法可以在不染色的情况下感知客观的生物分子，并将这种方法应用于识别分子的情况。出于目的，我们专注于组织中的非线性光学效应，该组织是由高强度非常短的持续时间脉冲激光（如飞秒激光）引起的。在实际研究中，我们开发了三个显微镜。荧光寿命成像显微镜在纳秒范围内，使用第二谐波生成光（SCG光）的胶原蛋白敏感显微镜和相干的抗STOKES相干罗马散射（CARS）显微镜。最成功的是SHG显微镜。基于直径为10微米的探头光点的反射型显微镜，以及胶原分子诱导的SHG光的极化测量，我们确定了各种人体组织中胶原蛋白纤维方向的分布。人类皮肤中产生的SHG数据表明，网状真皮具有胶原蛋白纤维的纠结结构，这与皮肤解剖学检查的结果是一致的。我们已经改进了SHG显微镜，使用con-focal光学和3D样品扫描阶段获得了3D解剖图像。使用这种显微镜，已经测量了人皮肤和骨骼中拼贴纤维方向的层状。另外，我们还测量了用汽车显微镜在红细胞周围透明质酸的分布。