Single-Cell Recording of Vesicle Release From Human Neuroblastoma Cells Using 1024-ch Monolithic CMOS Bioelectronics.
Single-Cell Recording of Vesicle Release From Human Neuroblastoma Cells Using 1024-ch Monolithic CMOS Bioelectronics.
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DOI：
10.1109/tbcas.2018.2861220
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发表时间：
2018-12
影响因子：
5.1
通讯作者：
Kim BN
中科院分区：
文献类型：
--
作者：
White KA;Mulberry G;Smith J;Lindau M;Minch BA;Sugaya K;Kim BN
人类神经母细胞瘤细胞SH-SY5Y常被用作研究帕金森病和黑质多巴胺释放的神经元模型，黑质是中脑区域，在运动控制中起重要作用。利用单个囊泡释放事件的安培单细胞记录，我们可以研究多巴胺释放的分子操作，并更好地了解神经系统疾病的机制。然而，单细胞分析的神经递质释放使用传统技术产生的结果非常低的通量。在本文中，我们将讨论一种单片集成CMOS传感器阵列，它具有低噪声性能，良好的时间分辨率，以及1024个并行通道，可以以单泡分辨率观察许多单细胞的多巴胺释放。我们的跨阻放大器的测量噪声电平分别为415、622和1083 fARMS，采样率分别为10、20和30 kS/s，无需额外滤波。后CMOS处理将1024个片上金电极（单个电极尺寸为15 μm × 15 μm）直接单片集成在CMOS器件中的1024个透阻放大器上。SU-8陷阱是在单独的电极上制造的，允许单细胞被询问，并拒绝多细胞团块。通过将SH-SY5Y细胞装入CMOS器件，同时记录76个细胞的多巴胺分泌。在42秒的测量中，总共监测了7147个单囊泡释放事件。该研究表明，CMOS器件具有在单细胞水平上记录囊泡分泌的能力，具有1024个平行通道，可以在单囊泡分辨率上提供多巴胺释放动力学的详细信息。
Human neuroblastoma cells, SH-SY5Y, are often used as a neuronal model to study Parkinson’s disease and dopamine release in the substantia nigra, a midbrain region that plays an important role in motor control. Using amperometric single-cell recordings of single vesicle release events, we can study molecular manipulations of dopamine release and gain a better understanding of the mechanisms of neurological diseases. However, single-cell analysis of neurotransmitter release using traditional techniques yields results with very low throughput. In this paper, we will discuss a monolithically-integrated CMOS sensor array that has the low-noise performance, fine temporal resolution, and 1024 parallel channels to observe dopamine release from many single cells with single-vesicle resolution. The measured noise levels of our transimpedance amplifier are 415, 622, and 1083 fARMS , at sampling rates of 10, 20, and 30 kS/s, respectively, without additional filtering. Post-CMOS processing is used to monolithically integrate 1024 on-chip gold electrodes, with an individual electrode size of 15 μm × 15 μm, directly on 1024 transimpedance amplifiers in the CMOS device. SU-8 traps are fabricated on individual electrodes to allow single cells to be interrogated and to reject multicellular clumps. Dopamine secretions from 76 cells are simultaneously recorded by loading the CMOS device with SH-SY5Y cells. In the 42-second measurement, a total of 7147 single vesicle release events are monitored. The study shows the CMOS device’s capability of recording vesicle secretion at a single-cell level, with 1024 parallel channels, to provide detailed information on the dynamics of dopamine release at a single-vesicle resolution.