Quick Start And A Bit More Of BLM Technique.

By Alexandr Chanturiya

In almost five years that passed form the first edition of “Quick Start...” I got a lot of questions from new adepts of BLM-ology on various issues they have to overcome in a thorny way of mastering The Art of BLM Making. A bit tired of writing same advises repeatedly I started to collect FAQ in order to add them to next edition of this book. When I got enough of those, I realized that I can add more than just one new chapter and added introduction to ion selectivity and membrane fusion. I also edited some passages in old chapters in order to make them more 21-st century (few people use chart recorders now).
This book is intended to help researches with little or no experience with BLM technique to get basic understanding of the method, facilitate selection of the components required to build the optimal BLM setup and to begin experiments in shortest possible time.

• Language: English
• Publisher: Alexandr Chanturiya
• Release date: Nov 23, 2017
• ISBN: 9781370412549

Alexandr Chanturiya

Александр Нодариевич Чантурия родился в 1955 г., в СССР в городе Марганец, что на Украине. Вырос в Николаеве, учился в университетах Днепропетровска и Киева. По специальности биофизик. В 1991-м поехал на четыре месяца в научную командировку в США. Задержался. Работал в NIH, Genetic Therapy, Futron, снова в NIH. Отредактировал и опубликовал воспоминания своего отца. Понравилось. Решил переквалифицироваться в писатели. Alexandr Nodarievich Chanturiya was born in Marganetz, USSR, Ukraine, in 1955. Grow up in Nikolaev. In 1972 entered Dnepropetrovsk State University and after two years left it for Kiev State University that finished in 1977 with diploma in biophysics. Few years later become PhD in biochemistry and worked happily in A.V. Palladin Institute of Biochemistry, Neurochemistry department. After a year in University College of London in 1990-1991 went to USA for four month but somehow managed to stay longer. Worked in NIH, Genetic Therapy, Futron, again in NIH. At some point got feeling that jobs with good ratio of $ to fun would not last forever and started his own company Eastern Scientific. Later decided to have even more fun and published several online worstsellers. Plan to become rich and famous soon.

Book preview

Foreword

In almost five years that passed form the first edition of Quick Start... I got a lot of questions from new adepts of BLM-ology on various issues they have to overcome in a thorny way of mastering The Art of BLM. A bit tired of writing same advises repeatedly I started to collect FAQ in order to add them to next edition of this book. When I got enough of those, I realized that I can add more than just one new chapter and added three more: Surface Tension, Ion Selectivity and Membrane Fusion. I also edited some passages in old chapters in order to make them more 21-st century (few people use chart recorders now). Also I got a feeling that my understanding of a and the usage is better now and replaced one with another wherever I think it possible.

Cover Image: Fusion of a liposome labeled with rhodamin PE and calcein with BLM. (Ref.#10)

Table of Contents

1. Introduction.

2. Equipment required.

3. General recommendations on setting up BLM workstation.

4. Making Mueller-Rudin BLM.

5. Making Montal-Mueller BLM.

6. Making horizontal BLM.

7. Examples of BLM experiments.

7A. Membrane lifetime measurements.

7B. Capacitance measurements.

7C. Surface tension measurements.

7D. Testing membrane activity of proteins or peptides.

7E. Potential dependencies of protein incorporation.

7F. Current/voltage dependencies.

7G. Measuring ion selectivity.

7H. Membrane fusion.

8. Appendix.

8A. Dimensional drawing of BLM chamber.

8B. BLM chamber cleaning and reassembling..

8C. Circuit Diagram of a basic BLM amplifier.

8D Troubleshooting.

9. Abbreviations.

10. References.

1. Introduction

More than 55 years passed since first artificial lipid membranes were successfully created. Originally suggested technique (Mueller et al., 1962) is still used at present and a number of modifications and improvements of this technique were introduced. Thousands of published papers confirm viability and value of this approach for exploring properties of membranes, membrane proteins, and membrane fusion. Indeed, it is hardly possible to name the other biological method that allows registering the effect of a single molecule. Another factor that contributed to a wide spreading of this technique is that experimenting with artificial lipid membranes or BLM is relatively simple and inexpensive. However, when BLM setup is hastily assembled and used by a person without practical knowledge of the method, the result could lead either to long and frustrating learning of the basics of a bilayer measurements or incredible discoveries that discredit both the method and the author, if published.

This publication mainly addressed to researches who have little or no experience with BLM technique. It is intended to provide basic understanding of the method, facilitate selection of components required to build the optimal BLM setup and help them to start experiments in shortest possible time.

2. Equipment required

The most time consuming part of setting up system for BLM experiments is gathering all the equipment and chemicals. This chapter describes necessary and optional components needed to start working with BLM.

Necessary components

Amplifier. Since electrical currents produced byBLM are in pico- nano-ampere range it is absolutely necessary to amplify this current up to the level compatible with registering devices and convert

current into a proportional