Johnson Brandon E, Brown Austin L, Goodman Miriam B
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA, USA.
J Vis Exp. 2008 Oct 22(20):964. doi: 10.3791/964.
Pressure-polishing is a method for shaping glass pipettes for patch-clamp recording. We first developed this method for fabricating pipettes suitable for recording from small (<3 m) neuronal cell bodies. The basic principal is similar to glass-blowing and combines air pressure and heat to modify the shape of patch pipettes prepared by a conventional micropipette puller. It can be applied to so-called soft (soda lime) and hard (borosilicate) glasses. Generally speaking, pressure polishing can reduce pipette resistance by 25% without decreasing the diameter of the tip opening (Goodman and Lockery, 2000). It can be applied to virtually any type of glass and requires only the addition of a high-pressure valve and fitting to a microforge. This technique is essential for recording from ultrasmall cells (<5 m) and can also improve single-channel recording by minimizing pipette resistance. The blunt shape is also useful for perforated-patch clamp recording since this tip shape results in a larger membrane bleb available for perforation.
压力抛光是一种用于制作膜片钳记录用玻璃微电极的方法。我们最初开发这种方法是为了制造适合从小型（<3μm）神经元细胞体进行记录的微电极。其基本原理与玻璃吹制相似，结合气压和热量来改变由传统微电极拉制仪制备的膜片微电极的形状。它可应用于所谓的软质（钠钙）玻璃和硬质（硼硅酸盐）玻璃。一般来说，压力抛光可使微电极电阻降低25%，而不会减小尖端开口的直径（古德曼和洛克里，2000年）。它几乎可应用于任何类型的玻璃，只需要增加一个高压阀并安装到微型熔炉上。这项技术对于从超小型细胞（<5μm）进行记录至关重要，并且还可以通过最小化微电极电阻来改善单通道记录。钝头形状对于穿孔膜片钳记录也很有用，因为这种尖端形状会形成一个更大的可用于穿孔的膜泡。
