Department of Botany, University of Washington, Seattle, Washington 98195.
Plant Physiol. 1991 Dec;97(4):1573-5. doi: 10.1104/pp.97.4.1573.
A method is described for the isolation of protoplasts (Pisum sativum, Phaseolus vulgaris, Avena sativa, Arabidopsis thaliana) in preparation for ion flux studies using patch clamp electrophysiology. Protoplasts that have been exposed to hydrolytic, cell wall degrading, enzymes for as little as 5 minutes form gigaseals (seal resistance higher than 10 giga Ohm) with the patch pipette with success rates greater than 40%. Sealing of these protoplasts is fast, averaging less than 2 minutes. This method yields high rates of gigaseal formation in a variety of tissues from both monocots and dicots and will enhance data collection in ion flux studies of plasma membranes of vascular plants.
描述了一种分离原生质体（豌豆、菜豆、燕麦、拟南芥）的方法，以便用膜片钳电生理学进行离子通量研究。原生质体在接触到水解酶、细胞壁降解酶后，只需 5 分钟即可与膜片钳形成千兆欧姆级别的巨封（电阻高于 10 千兆欧姆），成功率大于 40%。这些原生质体的密封速度很快，平均不到 2 分钟。该方法可在单子叶植物和双子叶植物的多种组织中产生高比例的千兆欧姆封接，将提高对植物质膜离子通量研究的数据收集效率。
