Abstract
Neurons are the primary structural unit of nervous systems. The basic structure of a typical neuron consists of a cell body, dendrites and an axon. Axons play a main role in the signal transduction to other connected neurons. Recently, researches of local translation in axons has become center stage. These researches need to use techniques allowing to physically separate the cell bodies and axons. Several such devices have been developed. In my thesis, I use a new device developed in our Lab which can be used to collect more axons and be fabricated easily. Rat hippocampus neurons were prepared to culture in this new device. This new device has a double layered structure. The first layer is a piece of PCTE membrane, and the second layer is a thin PDMS (Poly dimethylsiloxane) sheet perforated by many round holes. I grow hippocampal neurons on the PCTE membrane. Most of the axons grow through the PCTE membrane into the second layer, PDMS membrane. On 17 day-in-vitro, many axons migrate the length of holes and grow on the opposite surface. Axons on PDMS surface were detected by using fluorescence immunostaining. I collect axons from the PDMS surface by using a rubber scraper. The content of axons in the collected sample is determined by comparing to the axon samples produced by using another lab-on-a-chip device also developed in the laboratory. Western blotting is also used to verify whether the axon sample has free contaminations from soma to dendrites. The results indicated that the new developed device in the laboratory can be used to harvest pure axons from cultured rat hippocampal neurons with a yield of axons around 4 fold higher than the earlier lab-on-a-chip device.