In vivo Eyes Development of Drosophila Embryo Studied by Three-dimensional Time-lapsed Imaging via Single Plane Illumination Microscopy (SPIM)

• Main Authors: Sih-Hua Pao, 鮑思樺
• Other Authors: Arthur Chiou
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/40157553934560448527

碩士 === 國立陽明大學 === 生醫光電研究所 === 101 === We have studied the development of Drosophila eyes in the embryo stage by Single Plane Illumination Microscopy (SPIM). In our SPIM setup, we aligned a pair of counter-propagating parallel beams with appropriate set of optical elements to produce a light sheet for fluorescence excitation of the sample. A compromise of lateral resolution, axial resolution, and the field of view with uniform fluorescence excitation has been achieved by proper combination of optical elements. The advantages of SPIM, including relatively high lateral resolution, capability of optically sectioning throughout the sample with high-speed data acquisition, and avoidance of (or reduction of) photo-bleaching was successfully demonstrated; besides, a special sample holding stage was designed and incorporated into the setup such that the sample can be rotated to improve the penetration depth and to enable multi-view image acquisition and subsequent image fusion. We have observed and recorded the dynamics of the CD gene expression during the course of Drosophila eyes development from stage 12 to stage 17 via time-lapse imaging of the embryogenesis of eye-antenna disc primordium (EADP) expressed with green fluorescent protein, and we also obtained preliminary results of the dynamics of en gene expression during the course of Drosophila embryo development after stage 14. Three-dimension time-lapse imaging of live sample (of Drosophila embryo) with good image quality was successfully demonstrated via multi-view reconstruction from two view angles (0-degree and 180-degree view angles).