单细胞囊泡释放分析

神经元之间的信息传递是依靠突触前膜神经递质释放实现的。在单细胞层次，递质囊泡尺寸微小，释放过程为毫秒级，因此发展高时空分辨的单细胞分析方法是关键。以单个PC-12细胞高钾刺激释囊泡释放过程为例，基于碳纤维微电极的单细胞安培法，可实现单细胞囊泡释放动力学的原位实时分析。本项目采用单细胞安培法记录单个PC-12细胞高钾刺激释囊泡释放过程。所用微电极内含5微米直径碳纤维，由P-2000微电极拉制仪拉制。单细胞层次单个递质囊泡释放数据使用700B膜片钳放大器记录。通过尼康TS100倒置显微镜和Sutter MP-225显微操作仪进行显微操作，通过Inbio RCP-2B加药系统给予高钾溶液刺激。以上与细胞直接接触的设备均置于法拉第笼中，用以屏蔽电磁干扰。碳纤维电极工作电压为780 mV，以Ag/AgCl为参比电极，700B信号低通滤波300 Hz，700B输出信号经Axon 1550A数模转换器转换并以10 KHz的采样频率记录。单细胞层次单个递质囊泡释放数据均使用Igor Pro 8软件分析，可分辨0.1 毫秒的释放信号变化；碳纤维电极接触测量细胞面积直径小于10 微米，可记录单个细胞的递质囊泡释放数据。本数据集包含了单细胞囊泡释放电化学分析示意图、单细胞囊泡释放电化学分析装置图、碳纤维微米电极照片、以及单细胞囊泡释放信号。

Information transmission between neurons relies on the release of neurotransmitters from the presynaptic membrane. At the single-cell level, neurotransmitter vesicles are tiny in size and their release occurs on the millisecond timescale, so developing single-cell analysis methods with high spatiotemporal resolution is critical. Taking the high potassium-stimulated vesicle release of a single PC-12 cell as an example, single-cell amperometry based on carbon fiber microelectrodes enables in situ real-time analysis of the kinetics of single-cell vesicle release. This project adopted single-cell amperometry to record the high potassium-stimulated vesicle release of individual PC-12 cells. The microelectrodes used consist of carbon fibers with a diameter of 5 μm and were fabricated using a P-2000 micropipette puller. Data on single neurotransmitter vesicle release events at the single-cell level were recorded using a 700B patch-clamp amplifier. Micromanipulation was performed using a Nikon TS100 inverted microscope and a Sutter MP-225 micromanipulator, and high-potassium solution stimulation was delivered via an Inbio RCP-2B drug application system. All equipment that directly contacts the cells was placed inside a Faraday cage to shield against electromagnetic interference. The carbon fiber electrode was operated at a working voltage of 780 mV, with an Ag/AgCl reference electrode. The signal from the 700B amplifier was low-pass filtered at 300 Hz, and the output signal of the 700B was converted by an Axon 1550A digital-to-analog converter and recorded at a sampling frequency of 10 kHz. Data on single neurotransmitter vesicle release events at the single-cell level were all analyzed using Igor Pro 8 software, which can resolve vesicle release signal changes with a temporal resolution of 0.1 ms; the contact area between the carbon fiber microelectrode and the measured cell has a diameter of less than 10 μm, enabling the recording of neurotransmitter vesicle release data from individual cells. This dataset includes schematic diagrams of electrochemical analysis for single-cell vesicle release, diagrams of the electrochemical analysis setup for single-cell vesicle release, photographs of carbon fiber microelectrodes, and single-cell vesicle release signal data.

搜集汇总

背景与挑战

背景概述

该数据集专注于单细胞层次囊泡释放的实时分析，利用碳纤维微电极和单细胞安培法实现毫秒级高时空分辨测量，以研究神经元信息传递机制。数据集包含电化学分析示意图、装置图、电极照片以及单细胞囊泡释放信号，适用于电化学分析学科领域。