Optical stimulation and imaging of functional brain circuitry in a segmented laminar flow chamber.
Optical stimulation and imaging of functional brain circuitry in a segmented laminar flow chamber.
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分段层流室中功能性脑电路的光学刺激和成像。
DOI：
10.1039/c2lc40689f
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发表时间：
2013
期刊：
影响因子：
6.1
通讯作者：
Hui,ElliotE
中科院分区：
文献类型：
--
作者：
Ahrar,Siavash;Nguyen,TransonV;Shi,Yulin;Ikrar,Taruna;Xu,Xiangmin;Hui,ElliotE
微流控技术正在成为研究脑片的有用工具，提供精确的化学因素输送以及强大的氧气和营养转运。然而，对基于电极的生理记录的持续依赖在物理访问以及可同时采样的部位数量方面造成了固有的限制。在本研究中，我们将微流体层流室与快速电压敏感染料成像和通过笼中谷氨酸的激光光刺激相结合，以绘制活的大脑切片中大片皮质区域的神经网络活动。我们发现，封闭的微流体室大大提高了神经信号光学测量的信噪比性能。这些光学工具也被用来表征设备内的层流界面，展示了小于100μm的功能边界宽度。最后，我们利用这个集成平台来研究发育中的小鼠海马区自发神经活动的信号传播机制。通过使用局部钙耗竭，我们为钙依赖的突触传递提供了证据。
Microfluidic technology is emerging as a useful tool for the study of brain slices, offering precise delivery of chemical factors along with robust oxygen and nutrient transport. However, continued reliance upon electrode-based physiological recording poses inherent limitations in terms of physical access, as well as the number of sites that can be sampled simultaneously. In the present study, we combine a microfluidic laminar flow chamber with fast voltage-sensitive dye imaging and laser photostimulation via caged glutamate to map neural network activity across large cortical regions in living brain slices. We find that the closed microfluidic chamber results in greatly improved signal-to-noise performance for optical measurements of neural signaling. These optical tools are also leveraged to characterize laminar flow interfaces within the device, demonstrating a functional boundary width of less than 100 μm. Finally, we utilize this integrated platform to investigate the mechanism of signal propagation for spontaneous neural activity in the developing mouse hippocampus. Through the use of localized Ca2+ depletion, we provide evidence for Ca2+-dependent synaptic transmission.