Levis R A, Rae J L
Department of Physiology, Rush Medical College, Chicago, Illinois 60612.
Biophys J. 1993 Oct;65(4):1666-77. doi: 10.1016/S0006-3495(93)81224-4.
Quartz has a dissipation factor of approximately 10(-4), which is an order of magnitude less than that of the best glasses previously used to fabricate patch pipettes; it's dielectric constant of 3.8 is also lower than that of other glasses. On the basis of these electrical characteristics it is expected that patch pipettes pulled from quartz tubing will produce significantly less noise than pipettes made from other glasses. Our work confirms these expectations and we describe theoretical and practical aspects of the use of quartz pipettes for single channel patch voltage clamp measurements. Methods for pulling quartz pipettes with a laser-based puller and coating them with low-loss elastomers are discussed, as are precautions that are necessary to achieve low noise recordings. We have shown that quartz pipettes can be pulled from tubing with outer diameter to inner diameter ratios as large as 3 and a method of applying heavy elastomer coatings all the way to the tip of pipettes is presented. Noise sources arising from the pipette and its holder are described theoretically, and it is shown that measured noise is in good agreement with such predictions. With low noise capacitive feedback electronics, small geometry holders, and thick-walled quartz pipettes coated with low-loss elastomers we have been routinely able to achieve noise of 100 fA rms or less in a 5-kHz bandwidth with real cell patches and a pipette immersion depth of approximately 2 mm. On occasion we have achieved noise as low as 60 fA rms in this bandwidth.
石英的耗散因数约为10^(-4),比之前用于制造膜片吸管的最佳玻璃的耗散因数小一个数量级;其3.8的介电常数也低于其他玻璃。基于这些电学特性,预计从石英管拉制的膜片吸管产生的噪声将明显低于由其他玻璃制成的吸管。我们的工作证实了这些预期,并且我们描述了使用石英吸管进行单通道膜片电压钳测量的理论和实践方面。讨论了使用基于激光的拉针仪拉制石英吸管并用低损耗弹性体进行涂层的方法,以及实现低噪声记录所需的注意事项。我们已经表明,可以从外径与内径之比高达3的管子拉制石英吸管,并提出了一种一直涂覆到吸管尖端的厚弹性体涂层方法。从理论上描述了吸管及其固定器产生的噪声源,并且表明测量的噪声与这些预测非常吻合。使用低噪声电容反馈电子设备、小尺寸固定器以及涂有低损耗弹性体的厚壁石英吸管,我们通常能够在5 kHz带宽内,对于真实细胞膜片且吸管浸入深度约为2 mm的情况下,实现均方根噪声为100 fA或更低。有时我们在该带宽内实现了低至60 fA均方根的噪声。