Pressure polishing: a method for re-shaping patch pipettes during fire polishing
Pressure polishing: a method for re-shaping patch pipettes during fire polishing
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DOI：
10.1016/s0165-0270(00)00224-7
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发表时间：
2000-07-31
影响因子：
3
通讯作者：
Lockery, SR
中科院分区：
文献类型：
--
作者：
Goodman, MB;Lockery, SR
膜片钳记录的分辨率受膜片移液器的几何和电学特性的限制。理想的全细胞膜片移液管具有钝的锥形尖端和低阻力。制作贴片移液器的最佳玻璃是低噪音、低电容的玻璃，如硼硅酸盐和铝硅酸盐玻璃。遗憾的是，几乎所有硼硅酸盐玻璃形成的移液管都具有尖锐的锥形尖端和相对较高的电阻。然而，在火抛光过程中，通过对移液管腔加压来重塑尖端是可能的，我们称之为“压力抛光”。我们发现这种技术适用于几乎任何类型的玻璃制成的移液器，包括厚壁铝硅酸盐玻璃。我们经常使用这种技术制作适合于全细胞膜片钳记录微小神经元（直径1-3 μ m）的移液器。我们的移液器由厚壁硼硅酸盐玻璃制成，尖端开口为亚微米，电阻< 10 M ω。类似的移液管可用于记录亚细胞神经元结构，如轴突、树突和树突棘。压力抛光在膜片钳应用中也很有用，因为膜片钳受益于使用钝尖端的移液器，例如穿孔贴片全细胞记录，低噪声单通道记录和需要内部灌注移液器的实验。(C) 2000 Elsevier Science B.V.版权所有
The resolution of patch-clamp recordings is limited by the geometrical and electrical properties of patch pipettes. The ideal whole-cell patch pipette has a blunt, cone-shaped tip and a low resistance. The best glasses for making patch pipettes are low noise, low capacitance glasses such as borosilicate and aluminasilicate glasses. Regrettably, nearly all borosilicate glasses form pipettes with sharp, cone-shaped tips and relatively high resistance. It is possible, however, to reshape the tip during fire polishing by pressurizing the pipette lumen during fire polishing, a technique we call 'pressure polishing.' We find that this technique works with pipettes made from virtually any type of glass, including thick-walled aluminasilicate glass. We routinely use this technique to make pipettes suitable for whole-cell patch-clamp recording of tiny neurons (1-3 mu m in diameter). Our pipettes are made from thick-walled, borosilicate glass and have submicron tip openings and resistances < 10 M Omega. Similar pipettes could be used to record from subcellular neuronal structures such as axons, dendrites and dendritic spines. Pressure polishing should also be useful in patch-clamp applications that benefit from using pipettes with blunt tips, such as perforated-patch whole-cell recordings, low-noise single channel recordings and experiments that require internal perfusion of the pipette. (C) 2000 Elsevier Science B.V. All rights reserved.