FACILITATED GIGA-SEAL FORMATION WITH A JUST ORIGINATED GLASS-SURFACE
FACILITATED GIGA-SEAL FORMATION WITH A JUST ORIGINATED GLASS-SURFACE
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DOI：
10.1007/bf00374265
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发表时间：
1994-07-01
影响因子：
4.5
通讯作者：
BENNDORF, K
中科院分区：
文献类型：
--
作者：
BOHLE, T;BENNDORF, K
描述了一种在贴片移液器中制备尖端的简单技术，该技术有利于千兆密封的形成。移液器由厚壁硼硅酸盐玻璃管（外径 2.0 毫米，内径 0.5 毫米）制成，吸头可以在浴中反复折断。移液管阻力相应地以3-20M Omega的步长从约80M Omega下降到约2M Omega(双浓缩Tyrode)。扫描电子显微镜显示断裂后获得的尖端相当平坦。这些破损的尖端特别适合膜片钳。在具有 Na+ 通道活性的 26 个斑块中，有 11 个心肌细胞中的 gi,oa-密封自发发育，即无需抽吸。在这些斑块中，与负压下形成的斑块相比，具有-40 mV去极化脉冲的平均电流幅度显着更高。结论是，自发密封的斑块最有可能是平面结构，并且Na+通道活性超过吸力诱导的斑块。
A simple technique of tip preparation in patch pipettes is described, which facilitates giga-seal formation. The pipettes were fabricated from thick-walled borosilicate glass tubing (external diameter 2.0 mm, internal diameter 0.5 mm) and the tips could be repeatedly broken in the bath. The pipette resistance correspondingly fell in steps of 3-20 M Omega from about 80 M Omega to about 2 M Omega (double concentrated Tyrode). Scanning electron microscopy showed that the tip obtained after breaking was fairly plain. These broken tips were especially appropriate for patch-clamping. In cardiac myocytes in 11 out of 26 patches with Na+ channel activity, gi,oa-seals developed spontaneously, i.e. without suction. In these patches the amplitude of the mean current with depolarizing pulses to -40 mV was significantly higher in comparison with patches formed under negative pressure. It is concluded that spontaneously sealed patches are most likely of planar configuration and the Na+ channel activity exceeds that in suction-induced patches.