We provide to the neuroscience research community unique, disposable platforms to sustain viability in vitro to cellular organization. Our neuron devices provide compartmentalization, fluidic isolation and improved cellular organization over traditio... TheXC-T500consistsof(2)500μmmicrogroovebarrierswitha500μmcentralcompartment.ThischipmakesitpossIBLetoculturecorticalneuronsintheleftandrightchamberswhileculturinghippocampalneurons,orothercombinationsofcells,inthemiddle.Thetriplecompartmentisalsosuitablefortransportstudieswheretheresearcherwishestotreatthemiddleoftheaxongrowinginthemiddlecompartmentwhileobservingeffectsatthegrowthconeorinthesomalcompartment.Neuronscanalsobeloadedinthecentralcompartmentanddifferenttreatmentsappliedtotheleftandrightcompartments. We provide to the neuroscience research community unique, disposable platforms to sustain viability in vitro to cellular organization. Our neuron devices provide compartmentalization, fluidic isolation and improved cellular organization over traditionally chaotic neuronal cell cultures.Xona Microfluidics’ founders and staff are experts in the field of neuroscience. Their goal is to make available to the research community the same tools and expertise they have developed for their own successful research and to develop innovative systems adapted to new and future needs.Xona Microfluidics’ proprietary manufacturing process is conducted under strict quality controls to assure consistent quality and reliability of the devices.Xona Microfluidics distributes and supports its products directly, as well as through qualified distributors, to hundreds of research organizations in over 20 countries.

XonaMicrofluidics LLC美国公司神经元突触轴突培养舱Microfluidic Chamber突触虽然是神经系统重要的功能单位,但针对突触的通用研究方法仍具有一定局限性,有待完善改进,为加深对突触发生、病变现象及机制的研究,本实验设计建立创新性研究方法,实现体外动态观察神经元突触结构的变化。【设计思路】本实验引入新型培养装置Microfluidic Chamber和跨突触结构重组GFP荧光蛋白对mGRASP实现神经元突触体外动态观察。Microfluidic Chamber有双侧培养空间,由微通道相通,仅能允许神经元轴突通过,同时根据流体动力学原理其双侧液体不会发生混合。mGRASP全称为mammalian GFP reconstitution across synapticpartners,包括Pre和Post两个蛋白,分别携带GFP蛋白的一部分,当二者距离约为20 nm左右时,能重组成GFP发出绿色荧光,反之则无荧光信号。Pre和Post通过neurexin和neuroligin固定表达在突触前膜和突触后膜上,利用突触间隙约为20 nm且小于正常细胞间距的原理,用绿色荧光特异性标记突触,体外动态观察突触结构。【实验内容】本实验在新型培养装置Microfluidic Chamber进行孕17天小鼠胎鼠皮层神经元原代培养,体外培养6~7 d,在两侧分别加入携带mGRASP Pre和Post基因的慢病毒感染神经元,基于Microfluidic Chamber良好的液相分隔性,能做到两侧分别表达Pre和Post蛋白,而不会出现二者同一细胞中共表达的情况。培养4周左右,表达Pre的神经元轴突穿过微通道与另一侧表达Post的神经元接触,形成突触后即可在突触间隙发出绿色荧光。可在培养基中加入神经营养因子或药物,体外动态观察生理、病理刺激对突触结构的影响。【材料】实验动物为孕17 dICR小鼠;Microfluidic Chamber购自Xona Microfluidics公司,货号为RD450;mGRASP基因由Dr. Jinhyun Kim惠赠。【可行性】在Microfluidic Chamber中进行神经元原代培养为已掌握技术;mGRASP蛋白已在细胞中表达,能检测到绿色荧光,证明实验可行。【创新性】由于突触结构的特性和通用标记方法大多具有细胞毒性,目前尚无较好办法实现神经元突触的体外动态观察,mGRASP作为荧光蛋白无明显细胞毒性,表达后仍可进行长期观察。mGRASP的应用必须基于Pre和Post独立表达,一旦在同一细胞中共表达即可发出绿色荧光,失去特异性标记突触的作用,因此mGRASP多通过定位注射的方式应用于在体实验,本实验设计创新性的利用Microfluidic Chamber良好的液相分隔性,实现了mGRASP的体外应用,进而动态观察突触结构。

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