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JoVE Encyclopedia of Experiments
Biology
趋电性测定:一种观察秀丽隐杆线虫运动的方法
趋电性测定:一种观察秀丽隐杆线虫运动的方法
Encyclopedia of Experiments
Biology
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Encyclopedia of Experiments Biology
Electrotaxis Assay: A Method to Observe Locomotion in C. elegans

趋电性测定:一种观察秀丽隐杆线虫运动的方法

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2,586 Views
03:22 min
April 30, 2023
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Transcript

- 在没有感觉刺激的情况下,秀丽隐杆线虫的运动通常是向前的,并被反转和转弯打断——导致方向改变的随机重定向作。秀丽隐杆线虫是电感受性的。也就是说,它们可以感应电场。当暴露于温和的直流电或直流电时,蠕虫沿负极或阴极的方向沿直线不间断的路径移动,这种行为称为趋电性。

为了更准确地观察蠕虫的运动,请将动物置于微流体装置中并施加直流电场。反转磁场的方向,蠕虫将转向相反的方向 - 朝向阴极的方向。施加交流电 AC,蜗杆将停止。磁场方向的交替变化压倒了蠕虫的神经运动系统,导致蠕虫无法固定。重新施加直流磁场,蠕虫将恢复向阴极前进。

在以下示例方案中,我们将看到一个设置,该设置演示了微流体设备中的趋电性测定。

- 通过将刚刚组装的微通道放在显微镜的 xy 可移动载物台上来开始此步骤,并将安装在监视器上的相机连接到监视器。将电源连接到微通道的电极。

确认微通道的电阻在 0.6 兆欧姆左右后,将微通道的输出管连接到一次性注射器上。然后将入口管的口浸入悬浮在 M9 生理缓冲液中的线虫溶液中。在注射器内施加负压,将液体吸入通道中。

当入口和出口管都装满时,断开注射器的连接,并通过调整管的相对高度以在通道中心放置一个蜗杆来静液压纵流量。然后将两根管子平放在相同的高度。为了在趋电实验期间切换蜗杆样品时保持微通道内的零流量,在将两个入口管调平到相同的高度后,如果仍有流量,请对管相对于彼此的高度进行小幅调整。

- 如果我们不确定流是否真的为零,我们可以通过改变电场的极性,然后评估蜗杆在转动时的线速度来诱导蜗杆运动的反转。

- 现在,将电源设置为适当的电压。激活电信号,并留出一分钟的预曝光时间,让蠕虫适应磁场,在此期间,蠕虫应开始向阴极移动。一分钟过后,开始录制。

实验完成后,从通道中取出所有液体和蠕虫。用去离子水冲洗腔室,并将设备放在 125 摄氏度的热板上晾干。

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