Dye loading with patch pipettes.
Dye loading with patch pipettes.
复制标题
使用贴片移液器加载染料。
DOI：
--
复制
发表时间：
2009
影响因子：
--
通讯作者：
A. Konnerth
中科院分区：
文献类型：
--
作者：
J. Eilers;A. Konnerth
该方案描述了通过贴片吸管将荧光探针装载到单个细胞中。膜片钳方法已成功地用于培养神经元、脑片和活体标本中的单细胞染料标记。这种负载技术可以使用广泛的染料。用于形态重建的标记(例如，荧光黄)；用于监测第二信使级联的离子敏感指示染料(例如，Fura-2)；以及用于荧光共振能量转移(FRET)、荧光相关光谱(FCS)和光漂白后荧光恢复(FRAP)研究的染料标记蛋白质，都适合于膜片钳负载。这项技术最广泛的应用是在钙离子成像方面。全细胞膜片钳记录代表了一种多功能的加载技术，允许在定量水平上结合电生理和光学测量。
This protocol describes the loading of individual cells with fluorescent probes via patch pipettes. The patch-clamp methodology has been successfully used for single-cell dye labeling in cultured neurons, brain slices, and in vivo preparations. A broad range of dyes can be used with this loading technique. Markers for morphological reconstruction (e.g., Lucifer yellow); ion-sensitive indicator dyes for monitoring second-messenger cascades (e.g., fura-2); and dye-labeled proteins for fluorescence resonance energy transfer (FRET), fluorescence correlation spectroscopy (FCS), and fluorescence recovery after photobleaching (FRAP) studies are all suitable for patch-clamp loading. The most widespread application of this technique has been for Ca(2+) imaging. Whole-cell patch-clamp recordings represent a versatile loading technique that allows combined electrophysiological and optical measurements at a quantitative level.