Example: biology

A Guide to Electrophysiology & Biophysics …

THE AXON Guide . A Guide to Electrophysiology &. Biophysics laboratory Techniques The Axon Guide 2500-0102 Rev. C. Molecular Devices (now part of MDS Analytical Technologies). The Axon Guide A Guide to Electrophysiology & Biophysics laboratory Techniques Copyright Third Edition Copyright 2008, MDS Analytical Technologies. All rights reserved. No part of this publication may be reproduced, transmitted, transcribed, stored in a retrieval system, or translated into any language or computer language, in any form or by any means, electronic, mechanical, magnetic, optical, chemical, manual, or otherwise, without the prior written permission of MDS. Analytical Technologies, 1311 Orleans Drive, Sunnyvale, California, 94089, United States of America. Trademarks CYBERAMP, DIGIDATA, GENECLAMP and PCLAMP are registered trademarks and AXOPATCH, AXOCLAMP and MULTICLAMP are trademarks of MDS Analytical Technologies.

The Axon Guide — 2500-0102 Rev. C THE AXON GUIDE A Guide to Electrophysiology & Biophysics Laboratory Techniques

Tags:

  Laboratory, Technique, To electrophysiology amp biophysics laboratory techniques, Electrophysiology, Biophysics

Information

Domain:

Source:

Link to this page:

Please notify us if you found a problem with this document:

Other abuse

Transcription of A Guide to Electrophysiology & Biophysics …

1 THE AXON Guide . A Guide to Electrophysiology &. Biophysics laboratory Techniques The Axon Guide 2500-0102 Rev. C. Molecular Devices (now part of MDS Analytical Technologies). The Axon Guide A Guide to Electrophysiology & Biophysics laboratory Techniques Copyright Third Edition Copyright 2008, MDS Analytical Technologies. All rights reserved. No part of this publication may be reproduced, transmitted, transcribed, stored in a retrieval system, or translated into any language or computer language, in any form or by any means, electronic, mechanical, magnetic, optical, chemical, manual, or otherwise, without the prior written permission of MDS. Analytical Technologies, 1311 Orleans Drive, Sunnyvale, California, 94089, United States of America. Trademarks CYBERAMP, DIGIDATA, GENECLAMP and PCLAMP are registered trademarks and AXOPATCH, AXOCLAMP and MULTICLAMP are trademarks of MDS Analytical Technologies.

2 These trademarks are not to be used in any type of promotion or advertising without written permission from MDS Analytical Technologies. All other trademarks or registered trademarks are the property of their respective owners. Disclaimer MDS Analytical Technologies reserves the right to change its products and services at any time to incorporate technological developments. This publication is subject to change without notice. Although this manual has been prepared with every precaution to ensure accuracy, MDS Analytical Technologies assumes no liability for any errors or omissions, nor for any damages resulting from the application or use of this information. Questions? Phone: +1-800-635-5577. +1-408-747-1700. Fax: +1-408-747-3603. Web: For additional offices, please see the contact information on the back cover of this publication.

3 2 The Axon Guide 2500-0102 Rev. C. Contents List of Figures ..ix List of Tables ..xiii Preface .. xv Introduction .. xvii Editorial ..xix Contributors ..xxi 1. Bioelectricity Electrical Potentials .. 1. Electrical Currents.. 1. Resistors and Conductors .. 3. Ohm's Law .. 5. The Voltage Divider .. 7. Perfect and Real Electrical Instruments .. 7. Ions in Solutions and Electrodes .. 9. Capacitors and their Electrical Fields .. 11. Currents Through Capacitors .. 12. Current Clamp and Voltage Clamp .. 14. Glass Microelectrodes and Tight Seals .. 16. Further Reading .. 17. 2. The laboratory Setup The In Vitro Extracellular Recording Setup .. 19. The Single-Channel Patch Clamping Setup.. 19. Vibration Isolation Methods .. 20. Electrical Isolation Methods .. 21. Radiative Electrical Pickup.. 21. Magnetically-Induced Pickup.

4 22. Ground-Loop Noise.. 22. Equipment Placement .. 23. List of Equipment .. 24. Further Reading .. 26. The Axon Guide 2500-0102 Rev. C i 3. Instrumentation for Measuring Bioeletric Signals from Cells Extracellular Recording .. 27. Single-Cell Recording .. 28. Multiple-Cell Recording .. 28. Intracellular Recording Current Clamp .. 29. Voltage Follower .. 29. Bridge Balance .. 30. Junction Potentials.. 32. Track .. 32. Current Monitor .. 33. Headstage Current Gain .. 35. Capacitance Compensation .. 35. Leakage Current.. 39. Headstages for Ion-Sensitive Microelectrodes.. 39. Bath Error Potentials .. 40. Cell Penetration: Mechanical Vibration, Buzz and Clear .. 45. Command Generation .. 45. Intracellular Recording Voltage Clamp .. 46. The Ideal Voltage Clamp .. 47. Real Voltage Clamps .. 47. Large Cells Two-Electrode Voltage Clamp.

5 48. Small Cells Discontinuous Single-Electrode Voltage Clamp .. 52. Discontinuous Current Clamp (DCC) .. 57. Continuous Single-Electrode Voltage Clamp (Whole-cell Patch Clamp) .. 58. Series-Resistance Compensation.. 59. Pipette-Capacitance Compensation .. 64. Whole-Cell Capacitance Compensation .. 66. Rupturing the Patch.. 68. Which One Should You Use: dSEVC or cSEVC? .. 69. Space Clamp Considerations .. 69. Single-Channel Patch Clamp .. 70. The Importance of a Good Seal .. 71. Resistor Feedback Technology .. 71. Capacitor Feedback Technology .. 74. Special Considerations for Bilayer Experiments .. 77. How Fast is Fast ? .. 78. Measurement of Changes in Membrane Capacitance .. 78. Seal and Pipette Resistance Measurement .. 79. Micropipette Holders .. 79. Current Conventions and Voltage Conventions .. 80. ii The Axon Guide 2500-0102 Rev.

6 C. Definitions.. 80. Whole-Cell Voltage and Current Clamp .. 81. Patch Clamp .. 82. Summary .. 83. References .. 84. Patch Clamp .. 84. Two-Electrode Voltage Clamp.. 84. Single-Electrode Voltage Clamp.. 84. Space-Clamp Considerations .. 84. Other .. 84. 4. Microelectrodes and Micropipettes Electrodes, Microelectrodes, Pipettes, Micropipettes and Pipette Solutions .. 85. Fabrication of Patch Pipettes .. 87. Pipette Properties for Single-Channel vs. Whole-Cell Recording .. 89. Types of Glasses and their Properties .. 89. Thermal Properties of Glasses .. 91. Noise Properties of Glasses .. 92. Leachable Components .. 94. Further Reading .. 94. 5. Advanced Methods in Electrophysiology Recording from Xenopus Oocytes .. 97. What is a Xenopus Oocyte? .. 98. Two-Electrode Voltage Clamping of Oocytes .. 99. Patch Clamping Xenopus Oocytes.

7 101. Further Reading .. 102. Patch-Clamp Recording in Brain Slices .. 102. The Cleaning technique .. 103. The Blind technique .. 106. Advantages and Disadvantages of the Two Methods of Patch Clamping Brain Slices .. 108. Further Reading .. 109. Macropatch and Loose-Patch Recording .. 109. Gigaseal-Macropatch Voltage Clamp .. 110. Loose-Patch Voltage Clamp .. 111. The Giant Excised Membrane Patch Method .. 117. Pre-Treatment of Muscle Cells.. 118. Pipette Fabrication .. 118. The Axon Guide 2500-0102 Rev. C iii Seal Formation.. 120. Further Reading .. 120. Recording from Perforated Patches and Perforated Vesicles .. 121. Properties of Amphotericin B and Nystatin .. 121. Stock Solutions and Pipette Filling .. 121. Properties of Antibiotic Partitioning .. 122. The Advantages of the Perforated-Patch technique .. 123. The Limitations of the Perforated-Patch technique .

8 123. Suggested Ways to Minimize the Access Resistance .. 124. Other Uses for Perforated Patches .. 125. Further Reading .. 128. Enhanced Planar Bilayer Techniques for Single-Channel Recording .. 129. Solving the Problems of High Resolution and Voltage Steps .. 130. Assembling a Bilayer Setup for High-Resolution Recordings .. 136. Further Reading .. 141. 6. Signal Conditioning and Signal conditioners Why Should Signals Be Filtered? .. 143. Fundamentals of Filtering .. 144. -3 dB Frequency.. 144. Type: High-pass, Low-pass, Band-pass or Band-reject (notch) .. 144. Order .. 144. Implementation: Active, Passive or Digital.. 144. Filter Function.. 144. Filter Terminology.. 145. - 3 dB Frequency .. 145. Attenuation .. 146. Pass Band.. 146. Stop Band .. 146. Phase Shift .. 146. Overshoot .. 146. Octave .. 146. Decade.. 146.

9 Decibels (dB) .. 147. Order .. 148. 10-90% Rise Time.. 149. Filtering for Time-Domain Analysis .. 149. Filtering for Frequency-Domain Analysis .. 151. Sampling Rate .. 152. Filtering Patch-Clamp Data .. 153. iv The Axon Guide 2500-0102 Rev. C. Digital Filters .. 153. Preparing Signals for A/D Conversion .. 155. Where to Amplify .. 155. Pre-Filter vs. Post-Filter Gain .. 156. Offset Control .. 157. AC Coupling and Autozeroing .. 158. Time Constant .. 160. Saturation .. 160. Overload Detection .. 160. Averaging .. 161. Line-Frequency Pick-Up (Hum) .. 161. Peak-to-Peak and rms Noise Measurements .. 161. Blanking .. 163. Audio Monitor Friend or Foe? .. 163. Electrode Test .. 164. Common-Mode Rejection Ratio .. 164. References .. 165. Further Reading .. 165. 7. Transducers Temperature Transducers for Physiological Temperature Measurement.

10 167. Thermistors .. 167. IC Temperature Transducers that Produce an Output Current Proportional to Absolute Temperature .. 168. IC Temperature Transducers that Produce an Output Voltage Proportional to Absolute Temperature .. 169. Temperature Transducers for Extended Temperature Ranges .. 169. Thermocouples .. 169. Resistance Temperature Detectors .. 170. Electrode Resistance and Cabling Affect Noise and Bandwidth .. 171. High Electrode Impedance Can Produce Signal Attenuation .. 171. Unmatched Electrode Impedances Increase Background Noise and Crosstalk .. 172. High Electrode Impedance Contributes to the Thermal Noise of the System .. 173. Cable Capacitance Filters Out the High-Frequency Component of the Signal.. 174. EMG, EEG, EKG and Neural Recording .. 174. EMG .. 174. EKG .. 175. EEG.. 175. Nerve Cuffs .. 175.


Related search queries