| Summary: | 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.
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