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2014 | Book

Gigaseal Formation in Patch Clamping

With Applications of Nanotechnology

Authors: Majid Malboubi, Kyle Jiang

Publisher: Springer Berlin Heidelberg

Book Series : SpringerBriefs in Applied Sciences and Technology

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About this book

This book presents an investigation of gigaseal formation using micro/nanotechnology. The aims of the book are twofold. First, it explains the mechanisms of gigaseal formation using the latest discoveries. Second, it provides practical techniques for frequent formation of high resistance seals. The formation of a high-resistance electrical seal, also known as a gigaseal, between a cell membrane and a glass micropipette tip is essential in patch-clamp experiments. Even though four decades have passed since the introduction of the patch-clamping technique by Neher and Sakmann, gigaseal formation remains an obstacle in developing the high-throughput ion channel screening systems required by the pharmaceutical industry. Here the authors share their latest methods for achieving gigaseal formation and describe techniques that are highly desirable at both research and industrial levels. Nanotechnology has been found to be a powerful tool for studying and modifying glass micropipettes and in tackling the problem of gigaseal formation.

Table of Contents

Frontmatter
Chapter 1. Introduction
Abstract
A gigaseal is a high-resistance seal in order of giga Ohms formed between cell membrane and patching tool. High-resistance seals are needed in order to be able to record high-quality data from cellular ion channels activities. Gigaseal formation has remained a mystery for decades, as no conditions can warrantee a gigaseal. Although the mechanism of gigaseal formation is not yet fully elucidated, studies have been successful in determining the important conditions that should be met for a gigaseal to form.
Majid Malboubi, Kyle Jiang
Chapter 2. Development of Patch Clamping
Abstract
Patch clamping was first introduced into biophysical studies by Neher and Sakmann [1] in 1976 and was soon expanded to many other fields such as biology and medicine. The technique not only allowed the detection of single-channel currents in biological membranes for the first time but also enabled higher current resolution, direct membrane patch potential control, and physical isolation of membrane patches [2]. The development of the patch clamp method was honoured with a Nobel Prize in 1991.
Majid Malboubi, Kyle Jiang
Chapter 3. Gigaseal Formation
Abstract
To have a good understanding of interactions and forces in gigaseal formation it is necessary to have a close look at the glass and membrane structures.
Majid Malboubi, Kyle Jiang
Chapter 4. Effect of Roughness on Gigaseal Formation
Abstract
A rough pipette tip in conventional patch clamping, or patching site in planar patch clamping prevents seal formation. In this chapter the effect of roughness on gigaseal formation is discussed.
Majid Malboubi, Kyle Jiang
Chapter 5. Effect of Hydrophilicity on Gigaseal Formation
Abstract
A membrane has both hydrophilic and hydrophobic components. The exact contribution of these components in seal formation is not clear. Hydrophilicity of the pipette (or patch site in planar patch clamping) is believed to be a prerequisite for gigaseal formation.
Majid Malboubi, Kyle Jiang
Chapter 6. Effect of Tip Size on Gigaseal Formation
Abstract
This chapter reports the research into the effect of tip size on gigaseal formation. The approach adopted in the research includes, first, investigation on how the tip size affects the patch clamping, then a study on the surface properties of pipettes with different tip sizes and compare the seal resistances obtained by them, and finally verification of the conclusions using experiments.
Majid Malboubi, Kyle Jiang
Chapter 7. Study of Glass Micropipettes from Tip Formation to Characterization
Abstract
In this chapter various aspects of glass micropipettes are studied, including mechanisms of tip formation, tip geometry, and effect of pulling parameters on surface roughness properties of glass micropipettes.
Majid Malboubi, Kyle Jiang
Metadata
Title
Gigaseal Formation in Patch Clamping
Authors
Majid Malboubi
Kyle Jiang
Copyright Year
2014
Publisher
Springer Berlin Heidelberg
Electronic ISBN
978-3-642-39128-6
Print ISBN
978-3-642-39127-9
DOI
https://doi.org/10.1007/978-3-642-39128-6