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Dynamics of T-Junction Solution Switching Aimed at Patch Clamp Experiments

机译:针对膜片钳实验的T型结溶液切换动力学

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摘要

Solutions exchange systems are responsible for the timing of drug application on patch clamp experiments. There are two basic strategies for generating a solution exchange. When slow exchanges are bearable, it is easier to perform the exchange inside the tubing system upstream of the exit port. On the other hand, fast, reproducible, exchanges are usually performed downstream of the exit port. As both strategies are combinable, increasing the performance of upstream exchanges is desirable. We designed a simple method for manufacturing T-junctions (300 μm I.D.) and we measured the time profile of exchange of two saline solutions using a patch pipette with an open tip. Three factors were found to determine the timing of the solution switching: pressure, travelled distance and off-center distance. A linear relationship between the time delay and the travelled distance was found for each tested pressure, showing its dependence to the fluid velocity, which increased with pressure. The exchange time was found to increase quadratically with the delay, although a sizeable variability remains unexplained by this relationship. The delay and exchange times increased as the recording pipette moved away from the center of the stream. Those increases became dramatic as the pipette was moved close to the stream borders. Mass transport along the travelled distance between the slow fluid at the border and the fast fluid at the center seems to contribute to the time course of the solution exchange. This effect would be present in all tubing based devices. Present results might be of fundamental importance for the adequate design of serial compound exchangers which would be instrumental in the discovery of drugs that modulate the action of the physiological agonists of ion channels with the purpose of fine tuning their physiology.
机译:溶液交换系统负责在膜片钳实验中应用药物的时间。生成解决方案交换有两种基本策略。如果需要进行缓慢的更换,则更容易在出口上游的管路系统内进行更换。另一方面,通常在出口下游进行快速,可重复的交换。由于两种策略都是可组合的,因此提高上游交换的性能是可取的。我们设计了一种简单的方法来制造T型结(内径300μm),并使用带有开口尖端的贴片移液器测量了两种盐溶液交换的时间曲线。发现了三个因素来确定溶液切换的时间:压力,行进距离和偏心距离。对于每个测试压力,发现时延和行进距离之间存在线性关系,表明其对流体速度的依赖性,该速度随压力而增加。尽管这种关系仍无法解释很大的可变性,但发现交换时间随延迟而二次方增加。随着记录移液器移离流中心,延迟和交换时间增加。随着移液器移至溪流边界附近,这些增加变得戏剧性。沿边界处的慢流体与中心处的快流体之间的行进距离进行的质量传输似乎有助于溶液交换的时间过程。在所有基于管道的设备中都会出现这种效果。目前的结果对于串行化合物交换剂的适当设计可能具有根本的重要性,这对于发现可调节离子通道生理激动剂作用以微调其生理学作用的药物至关重要。

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