Perforated patch recording with β-escin
Perforated patch recording with β-escin
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DOI：
10.1007/s004240050737
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发表时间：
1998-11-01
影响因子：
4.5
通讯作者：
Palade, P
中科院分区：
文献类型：
--
作者：
Fan, JS;Palade, P
对于某些研究人员来说，用制霉菌素、两性霉素 B 和短杆菌肽进行穿孔贴片记录可能比其他研究人员更困难。此外，在此类实验中很难将染料等低分子量物质引入细胞质中。我们已经确定，β-七叶皂苷是水溶性较差的离子载体的一种方便、易于使用的替代品。端口描述了离子载体的等待时间长和不便，离子载体首先需要溶解在非水溶剂中，然后再溶解在移液器溶液中。其他方法已用于透化细胞，包括洋地黄皂苷和皂苷，它们可以保护细胞内膜。 β-七叶皂苷是一种皂苷衍生物，最初用于使平滑肌透化[4]，据报道可形成对高分子量物质具有相当渗透性的孔洞[5]。我们在这里报告我们使用 β-七叶皂苷作为穿孔贴片记录的替代品的非常积极的经验，特别是当需要通过贴片吸管将指示剂或其他物质引入细胞进行长时间实验时。
Perforated patch recording with nystatin, amphotericin B and gramicidin can be more difficult in the hands of some investigators than others. In addition, it is difficult to introduce low molecular weight substances such as dyes into the cytoplasm in such experiments. We have determined that beta-escin represents a convenient, easy-to-use alternative to less water-soluble ionophores. ports describing long waiting times and inconvenience of working with ionophores that first need solubilization in nonaqueous solvents prior to dissolution in the pipette solution.Other methods have been used to permeabilize cells, including digitonin and saponin, which spare intracellular membranes. beta-escin, a saponin derivative used initially to permeabilize smooth muscle [4], has been reported to form holes quite permeable to high molecular weight substances [5]. We report here our very positive experiences with beta-escin as an alternative for perforated patch recording, particularly when there is a desire to introduce indicators or other substances into cells via the patch pipette for long-duration experiments.