Myocyte contraction visualized with lock-in amplified second-harmonic generation microscopy

• Main Author: 張嘉仁
• Other Authors: 廖奕翰
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/25547195441633534174

碩士 === 國立交通大學 === 分子科學研究所 === 99 === Optical microscopy, especially laser scanning microscopy, has been a powerful tool for biomedical research. Concerning laser scanning optical microscopy, a two-dimensional image is constructed by scanning the focus of a laser beam across a fixed sample or vice versa. The frame rate of the former is larger, but the latter allows incorporation of techniques such as lock-in amplification or photon counting to improve signal-to-noise ratio. We have developed a novel lock-in amplified laser scanning second-harmonic generation (SHG) microscopy system, by firstly integrating a pulsed laser into a commercial confocal microscope, and secondly incorporating modulation-demodulation technique into the system. The signal-to-noise ratio is significantly improved while maintaining fast frame rate. The thesis first details the setup of the system including the setting of experiment parameters, and follows with demonstration of the system to image the contraction of two distinct kinds of myocytes: electrically stimulated skeletal muscle cell line and spontaneously contracting chick embryo cardiomyocytes. We show that we can follow the contracting-resting cycle of either electrically induced contraction of skeletal muscles or spontaneous contraction of primary cardiomyocytes. Through image analysis, we can determine the change of sarcomere length during electrically stimulated or spontaneous contraction and examine the effect of hormone on sarcomere contractions.