Growth of neurospheres in a microfluidic device characterized with selective plane illumination microscopy

• Main Authors: Rui Xiang, 向睿
• Other Authors: Chau-Hwang Lee
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/64382508587472182686

碩士 === 國立陽明大學 === 生醫光電研究所 === 102 === Abstract To observe and culture the 3-D biological sample is always big challenge in biological experimints.3-D biological sample is closer to real condition then 2-D. In our study, we used SPIM(selective plane illumination microscopy) which is a type of light sheet microscope to research the growth of neurospheres. neurospheres is a 3-D functional biological sample which are aggregated by a lot of single neural stem cells,and neurospheres have ability of self-renew,proliferationand also will differentiated to neuron,astrocytes and oligodendrocytes. SPIM provide a low phototoxity,fast image acquisitionand 3-D reconstruction by scanning Z axis. Because neural stem cells were cultured to a 3-D functional neurospheres,the proliferation of neorosphere is the basic of cell culture in 3-D environment.The feature of light sheet microscopy is that illumination light will in the vertical direction with detection objective,so we developed a microfluidics device which can provide a micro environment for neurospheres.In the end, we observed the growth of cell in the 3 Days in the device and took the cell counting,demonstrated the relationship between the single neural stem cell and neurospheres. The thickness of beam waist is a key parameter of SPIM,we improve the illumination section by thinning the thickness of beam waist,promote the z resolution of system.The SPIM will be applied in the smaller scale biological sample.