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Development of Measurement System for Molecular Motion in Plasma Membrane using Pico-second Time-resolved Microscopic Photometry

利用皮秒时间分辨显微光度法开发质膜分子运动测量系统

基本信息

批准号：
07557004
负责人：
ARAISO Tsunehisa
金额：
$ 1.09万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

Time-resolved fluorescence depolarization method is a powerful tool to measure molecular motion of biological membranes. Usually this type of the measurement has been applied to membrane or cell suspension, and so the results were averaged values of the ensemble of molecules in the membranes or cells. However physiologically important changes of the molecular motions in the membranes occur at local areas. In this work, we use a microscope to develop the fluorescence depolarization method to detect the molecular motion at a local area with micrometer resolution. A pulsed Ar-dye laser was introduced to the sample membrane located on a microscope stage for excitation. The incident angle was variable. This function will be useful to detect the orientation angle of the molecules in membrane. Decay of the polarized fluorescent light passed through the objective lens was detected by a single photon counting technique. With this system, we could measure the anisotropic fluorescence decay from a single molecular layr of stearic acid containing 1 mol% of DPH-propionate in a 10 square micrometer area. This suggests that this system can be applied to measure molecular motion in a local area of cell membrane.

时间分辨荧光去极化法是测量生物膜分子运动的有力工具。通常这种类型的测量已应用于膜或细胞悬浮液，因此结果是膜或细胞中分子集合的平均值。然而，重要的生理变化的分子运动在膜发生在局部区域。在这项工作中，我们利用显微镜开发了荧光去极化方法，以微米分辨率检测局部区域的分子运动。将脉冲ar染料激光引入到位于显微镜台上的样品膜上进行激发。入射角是可变的。该函数可用于检测膜中分子的取向角。用单光子计数技术检测了通过物镜的偏振光的衰减。利用该系统，我们可以在10平方微米范围内测量含有1mol %丙酸dph的硬脂酸单分子层的各向异性荧光衰减。这表明该系统可用于测量细胞膜局部区域的分子运动。