neuromuscular junc lab manual

neuromuscular junc lab manual - Neuromuscular Junction Lab...

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Neuromuscular Junction Lab Manual © University of Minnesota Version: July, 2009
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Table of Contents 1 INTRODUCTION. .............................................................................................................................. 3 1.1 COURSE OVERVIEW. ..................................................................................................................... 3 1.2 PREVIEW OF EXPERIMENT ACTIVITIES . ......................................................................................... 4 1.3 NOTEBOOK. .................................................................................................................................. 5 1.4 YOUR RESPONSIBILITIES. .............................................................................................................. 5 1.5 KEEPING YOUR SANITY . ............................................................................................................... 5 1.6 HELP US IMPROVE THIS MANUAL. ................................................................................................. 6 2 EQUIPMENT AND INSTRUMENTATION . .................................................................................. 7 2.1 OVERVIEW OF NEUROPHYSIOLOGY EQUIPMENT . .......................................................................... 7 2.2 THE NMJ EQUIPMENT RACK. ........................................................................................................ 7 2.3 OSCILLOSCOPE BASICS. ................................................................................................................ 9 2.4 PULSE STIMULATOR BASICS . ........................................................................................................ 9 2.5 RECORDING ELECTRODE AND AMPLIFIER BASICS . ........................................................................ 9 2.6 COMPUTER-BASED EXPERIMENT CONTROL. .................................................................................. 9 3 TECHNIQUES, EXERCISES AND EXPERIMENTS. ................................................................. 10 3.1 GETTING TO KNOW THE LAB. ...................................................................................................... 10 3.2 CABLES AND CONNECTORS . ....................................................................................................... 10 3.3 GETTING TO KNOW YOUR OSCILLOSCOPE. .................................................................................. 11 3.4 WORKING THE STIMULATOR . ..................................................................................................... 17 3.5 USING THE ELECTRODE PREAMPLIFIER. ...................................................................................... 22 3.6 USING MICROSCOPES. ................................................................................................................. 24 3.7 SIMULATED CELL . ...................................................................................................................... 25 3.8 CHECKING ELECTRODE IMPEDANCE . .......................................................................................... 26 3.9 MAKING A GLASS MICROELECTRODE . ........................................................................................ 28 3.10 PITHING THE FROG . .................................................................................................................... 32 3.11 DISSECTING THE CUTANEOUS PECTORALIS MUSCLE. .................................................................. 33 3.12 MEASURING MEMBRANE POTENTIAL. ......................................................................................... 37 3.13 MINIATURE END PLATE POTENTIALS (MEPPS). .......................................................................... 40 3.14 EFFECT OF K+ CONCENTRATION ON MEMBRANE POTENTIAL. ..................................................... 42 3.15 STIMULATING THE NERVE FIBER WITH A SUCTION ELECTRODE . ................................................. 45 3.16 MOTOR UNIT ACTION POTENTIALS . ............................................................................................ 47 3.17 END PLATE POTENTIALS. ............................................................................................................ 50 3.18 MEASURING SARCOMERE LENGTH . ............................................................................................ 52 3.19 FORCE MEASUREMENTS . ............................................................................................................ 54 3.20 FORCE SENSOR SETUP. ................................................................................................................ 55 3.21 MUSCLE TWITCH FORCE. ............................................................................................................ 57 4 YOUR RESEARCH PROJECT. ..................................................................................................... 61 4.1 PURPOSE . ................................................................................................................................... 61 4.2 DELIVERABLE . ........................................................................................................................... 61 4.3 PRESENTATION TIPS . .................................................................................................................. 62 4.4 RESEARCH PROJECT SUGGESTIONS. ............................................................................................ 62 APPENDIX A TEK SCOPE REFERENCE . ................................................................................... 63 APPENDIX B SAMPLE PLOTS. ..................................................................................................... 67 APPENDIX C STIMULATOR CONTROLS. ................................................................................. 69 APPENDIX D FROG RINGER'S SOLUTIONS. ............................................................................ 71 1
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APPENDIX E NOISE!. ...................................................................................................................... 73 APPENDIX F BASIC EXPERIMENT DESIGN AND STATISTICS . ......................................... 74 APPENDIX G FITTING A STRAIGHT LINE TO DATA USING EXCEL. ............................... 75 APPENDIX H BIPOLAR HOOK ELECTRODES. ........................................................................ 77 APPENDIX I FIXING SUCTION ELECTRODES. ........................................................................... 79 APPENDIX J MEASURING ELECTRODE AND MEMBRANE IMPEDANCE. .......................... 80 APPENDIX K CHANGING SETTINGS ON THE SUTTER MICROPIPETTE PULLER. ...... 84 2
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1 Introduction 1.1 Course overview Welcome to the Neuromuscular Junction short course. The purpose of this course is to provide you with the background and skills needed to understand basic neurophysiology with an emphasis on nerves, skeletal muscle and the neuromuscular junction. During the week, there will be a few foundation lectures that cover the basics of nerve, muscle and muscle disease, and classic electrophysiology measurement methods. The primary purpose of the course, however, is for you to conduct hands-on experiments using an isolated frog nerve-muscle preparation. Lecture topics for the week will include: Equipment in the electrophysiology lab Biophysical Basis of Membrane Potentials Neuromuscular Signal Transmission Muscle Contraction plus several lectures on groundbreaking research happening at the University of Minnesota. The experiment lab stations contain state-of-the-art equipment for exciting nerves and recording the resulting activity in membrane, nerve and muscle. Enrollment in the course is kept small to ensure there will only be two or three students per station. This means you can learn by doing rather than learn by reading or learn by watching.
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neuromuscular junc lab manual - Neuromuscular Junction Lab...

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