Integrating multi-unit electrophysiology and plastic culture dishes for network neuroscience
Integrating multi-unit electrophysiology and plastic culture dishes for network neuroscience
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DOI：
10.1039/b802165a
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发表时间：
2008-01-01
期刊：
影响因子：
6.1
通讯作者：
Claverol-Tinture, Enric
中科院分区：
文献类型：
--
作者：
Morales, Ricardo;Riss, Michael;Claverol-Tinture, Enric
培养神经元的电生理特性对于药物发现、安全药理学和神经科学的基础研究至关重要。提供低成本、低技术复杂性和朝向体外神经元上的高通量电生理学的可扩展性的潜力的技术将是有利的，特别是对于筛选目的。在这里，我们描述了一种塑料培养基，支持低复杂性的多单元松散补丁记录和刺激发展中的网络，同时保持与低成本和大规模生产兼容的可制造性。我们的混合聚二甲基硅烷（PDMS）上的聚苯乙烯结构包括室（直径6毫米）和微通道（25 μ m × 3.7 μ m 1毫米）作为基板嵌入式记录移液管。由于几何约束，胞体被镀覆并保留在腔室中，并且它们的突起沿着微通道生长，有效地建立了松散的补丁配置而无需人为干预。我们证明，现成的电压钳，电流钳和细胞外放大器可以用来记录和刺激多单位活动的帮助下，我们的菜。尖峰高达50 pA的电压钳和300 μ V的电流钳模式记录在稀疏和突发的活动模式的特征1周龄海马文化。此外，尖峰排序采用主成分分析（PCA）证实，单个微通道支持多个神经元的记录。总体而言，这项工作提出了一种策略，为传统文化塑料器皿赋予额外的功能，以实现具有成本效益的网络电生理学。
The electrophysiological characterisation of cultured neurons is of paramount importance for drug discovery, safety pharmacology and basic research in the neurosciences. Technologies offering low cost, low technical complexity and potential for scalability towards high-throughput electrophysiology on in vitro neurons would be advantageous, in particular for screening purposes. Here we describe a plastic culture substrate supporting low-complexity multi-unit loose-patch recording and stimulation of developing networks while retaining manufacturability compatible with low-cost and large-scale production. Our hybrid polydimethylsilane (PDMS)-on-polystyrene structures include chambers (6 mm in diameter) and microchannels (25 mu m x 3.7 mu m 1 mm) serving as substrate-embedded recording pipettes. Somas are plated and retained in the chambers due to geometrical constraints and their processes grow along the microchannels, effectively establishing a loose-patch configuration without human intervention. We demonstrate that off-the-shelf voltage-clamp, current-clamp and extracellular amplifiers can be used to record and stimulate multi-unit activity with the aid of our dishes. Spikes up to 50 pA in voltage-clamp and 300 mu V in current-clamp modes are recorded in sparse and bursting activity patterns characteristic of 1 week-old hippocampal cultures. Moreover, spike sorting employing principal component analysis (PCA) confirms that single microchannels support the recording of multiple neurons. Overall, this work suggests a strategy to endow conventional culture plasticware with added functionality to enable cost-efficient network electrophysiology.