BIO325 Neurophysiology

Spring 2020

Laboratory Guide

LABORATORY MANUAL

Spring 2020

 

 

KEY   PASSIVE MEMBRANE   SYNAPSES

  Set Z - No Lab Writeup  

 

  Set A - Organismal Recording Labs 

  Set B - Electronic Simulation Labs 

  Set C - Computer Simulation Labs 

  Lab 6: Computer Simulations of Resting Potentials and Passive Spread  

Lab 14: Electronic Simulations  of

Electrical Synapses

 

Lab 7:Resting Potentials in

Crayfish Muscle Cells

 

Lab 15: Postsynaptic Potentials in

the Crayfish NMJ

INTRODUCTION  

Lab 8: Electronic Simulations of

Neuronal Cable Properties

 

Lab 16: Computer Simulations of

Postsynaptic Potentials

Lab 1: Electrophysiological

Instrumentation

  ACTION POTENTIALS  

Lab 17: Neuroplasticity in the

Crayfish NMJ

Lab 2: PowerLab

Tutorial

 

Lab 9: Electronic Simulations of

Action Potentials

  SENSORY PROCESSING
ELECTROPHYSIOLOGY  

Lab 10: Computer Simulations of

 a Voltage-Clamp Experiment

 

Lab 18: Electroretinogram (ERG)

of the Fly Eye

Lab 3: Recording from

Weakly Electric Fish

 

Lab 11: Computer Simulations of

Action Potentials

 

Lab 19: Crayfish Stretch

Receptors

Lab 4: Recording from

the Crayfish IIIs Root

 

Lab 12: Action Potentials

in the Snail Brain

  MOTOR SYSTEMS

Lab 5: Resistors, Capacitors,

and RC Circuits

 

Lab 13: Computer Simulations of

Action Potential Propagation

 

Lab 20: Motor Activity in

Crayfish Abdominal Nerves

 

TENTATIVE LABORATORY SCHEDULE

 

    CLASS/LAB EXERCISES AND DISCUSSIONS

W

Date

 

Monday

Wednesday

Wednesday
Lab

Friday

1

Jan 13-17

 

Course Intro

Neurons

Lab 1 - Electrophys Instrumentation

Lab 2a - PowerLab Tutorial

2

Jan 20-24

 

MLK Holiday

Lab 2b - PowerLab Tutorial

Lab 3-  Electric Fish  [A]

Electric Circuits

3

Jan 27-31

 

Lab 5a - DIYN RC Circuits

Lab 5b -  DIYN RC Circuits

Lab 4 -  Crayfish IIIs [A]

Membrane Potentials

& Currents

4

Feb 3-7

 

Estimates of the Resting Potential

Equivalent Circuit
- Compartment

Lab 4 -  Crayfish IIIs  [A]

Lab 6a -  NIA  Resting Potentials  [C]

5

Feb 10-14

 

Equivalent Circuit

- Distributed

Lab 6b -  NIA  passive Spread  [C]

Lab 7 - Crayfish Muscle RPs  [A]

Lab 8a -  DIYN Cable Properties  [B]

6

Feb 17-21

 

Lab 8b - DIYN Cable Properites  [B]

Review/Catchup

Midterm I

Action Potentials

7

Feb 24-28

 

Lab 9a - DIYN AP Simulation  [B]

Lab 9b -  DIYN AP Simulation  [B]

Lab 7 - Crayfish Muscle RPs  [A]

AP Mechanism

H-H Voltage Clamp

8

March 2-6

 

Lab 10a - NIA Voltage Clamp  [C]

Lab 10b - NIA Voltage Clamp  [C]

Lab 12 - Snail Brain Recording  [A]

Lab 11a - NIA Action Potentials   [C]

  March 9-13   Spring Break Spring Break

Spring Break

Spring Break

9

March 16-20

 

Lab 11b - NIA Action Potentials  [C]

On Beyond H-H

Lab 12 - Snail Brain Recording  [A]

Action Potential Propagation Mechanism

10

March 23-27

 

Lab 13a - NIA AP Propagation  [C]

Lab 13b - NIA AP Propagation  [C]

Lab 16 - Crayfish NMJ  [A]

Synapses - Chemical vs. Electrical

11

March 30

 - April 3

 

Lab 14a - NIA PSPs  [C]

Lab 15b - DIYN Elect Synapses   [B]

Lab 16/17 - Crayfish NMJ/Neuroplast  [A]

Lab 15b - DIYN Elect Synapses   [B]

12

April 6-10

 

Summation & Neuronal Integration

Review/Catchup

Midterm II

Easter Break

13

April 13-17

 

Lab 14b - NIA PSP Integration  [C]

Neuroplasticity Mechanisms

Lab 17 - Crayfish Neuroplasticity  [A]

Spike Train Analysis

14

April 20-24

 

Sensory Coding

Student Scholarship Day

Student Scholarship Day

Sensory Processing

15

April 27
 - May 1

 

Field Potentials

Sensorimotor Integration

Lab 18 - Crayfish Stretch Receptor  [A]

Review/Catchup

16

May 4-6

 

Review/Catchup

Review/Catchup

Lab 19 - Fly ERG [A]

 

Finals Week

May 7

May 8-12

 

Thursday Reading Day

Final Exam DATE and TIME TBD*

LAB WRITEUPS:

[A]  -  4 of 8

[B]  -  2 of 3

[C]  -  2 of 5

 *The lab portion of the FInal Exam is a 1 on 1 verbal final to be scheduled with the instructor during the final week of class.  It will focus on laboratory instrumentation and methods.

 

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OVERVIEW AND ASSIGNMENTS

 

As you have no doubt noticed, this is a uniquely lab-centered course.  50% grade is based directly on the products of your successful participation in the laboratory exercises.  The remaining 50% of your grade is based indirectly on what you learn; primarily during the laboratory exercises.  It will be essential in this course to keep up with the readings and computer simulations, come to each class/lab session completely prepared, participate fully throughout each session, keep careful track of your data, and turn in your laboratory assignments in a timely manner.

Laboratory Participation

If you reliably show up for labs, work well with your lab partners, and contribute your fair share, you should learn and understand enough to do fairly well in the course.  If you skip labs, don't pull your weight, sit around and watch others work, whine a lot, and generally let down your lab partners, you may find yourself entirely dependent upon their (probably exhausted) goodwill to survive the course at all.

Data Sheets

 

With luck there will be nineteen laboratory exercises in this course.  Three of the early exercises are Introductory Labs, designed primarily as learning experiences.  Although you will not be turning in any results from these labs and will not be directly graded on your participation in them, it is important that you complete these labs.  If you try to skip or gloss over these labs, you will be missing essential skills and concepts that will be essential for the graded labs.  The midterm and final exams will draw on your understanding of these introductory labs.  In particular, the verbal lab final will rely heavily on material learned (or not learned) in these labs.

 

The remaining sixteen laboratories are divided into four categories, color-coded above:

 

Set A - Organismal Recording Labs - Eight labs involve surgical preparation and recording of neural or bioelectrical signals from real biological organisms, most commonly crayfish.  You will need to turn in Data Sheets for four of these nine labs.

 

Set B - Electronic Simulation Labs - Three labs involve setup and recording recording from hardware electronic models which emulate specific aspects of neuronal structure and behavior.  You will need to turn in Data Sheets for two of these.

 

Set C - Computer Simulation Labs - Five labs involve working through sets of computer simulations of neuronal behavior and generating representative traces or plots from the output of those simulations.  You will need to turn in Data Sheets for two of these.

 

Each Data Sheet is a concise set results from a single laboratory exercise.  These results will be in the form of annotated recording "traces", data tables, well-labeled graphs, and occasionally answers to specific questions.   The exact contents of each data sheet will be specified in the guide to that particular lab.  Each Data Sheet is due within two calendar weeks of the completion of that particular lab exercise.

 

Laboratory Final Examination

 

The laboratory portion of the final exam will be a one-on-one verbal examination which you will schedule with the instructor.  As specified in the syllabus, the exam will focus the methodology of the course, especially that pertaining to surgical procedures and electrophysiological recording/analysis. 

 

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POWERLAB STATIONS AND COMPUTER ACCESS

     

PowerLab Stations and Computer Access

 

      This laboratory centers physically around four computer-based electrophysiology recording and analysis systems.  Each of these consists of a PC, a multi-channel PowerLab box, and PowerLab software.  The computers are named Margaret, Julia, Mary, and Bebbie (in honor of four women who contributed significantly to establishing and equipping this laboraroty space).  These computers are here for your use IN THIS COURSE.  In general, they will be on virtually 24-hours a day.  The only significant downtime will be when they are being physically moved from room to room.  Each computer has its own color printer and is connected to the web via a wireless hub. 

 

You can use these computers any time the room is free, and store your data on the hard drives, as long as you obey the following rules:

 

1)   Use only the PhysStudent login. This login is password protected.  DO not share the password with other students who are not in this course.  DO NOT USE THE INSTRUCTOR LOGIN.

2)   To gain network access you may have to use the wesleyanne login and cccr pasword.

3)   Please store your files in an orderly manner, within appropriate and well-labeled folders.  Back up your files on your own floppy or personal computer hard drive as soon as possible after creating them.  Be advised that any files left on the computer “desktop” may be thrown away at my discretion.

4)   Don't change the desk-top pattern.  I like it the way it is.

5)   Don’t change anything about the system, display, or defaults.

6)   Don’t download any files from the web to the computer.

7)   Don’t use the computer to check your e-mail.

8)   Don't use the computers or printers for coursework other than in the Biology or Neurosceince programs.

9)   Don't mess with other people's files, rename the hard drives, or change other setups.

10) Close all files and applications when you are finished with them.

11) Report any computer housekeeping or etiquette problems to the instructor as soon as possible, especially any issues related to the printers.

 

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LABORATORY  SAFETY  PRECAUTIONS

 

Laboratory study and investigation can be both fun and rewarding; however it can involve a certain amount of risk due to the nature of the equipment (e. g.  glassware, hot plates, scalpels, high voltage power supplies), specimens (e. gAscaris, bacteria, mammalian blood) and/or chemicals (e. g.  formalin, hydrochloric acid, sodium hydroxide, neurotoxins) used.  In order to minimize the chances of accidents and injury, the following general precautions must be followed in all scientific laboratories:

 

1.   NO SMOKING, EATING or DRINKING at any time.

 

2.   Shoes must be worn at all times (sandals are not shoes).

 

3.   Know the location of fire extinguishers, eye-wash stations, fire blankets, safety showers, first aid stations, and containers for broken glass.

 

4.   In the case of defective or broken equipment:

            -- Do not attempt to unplug frayed electric cords yourself;

            -- Do not attempt to clean up any broken glassware yourself;

            -- Report all defective equipment to the instructor.

 

5.   Be sure that electrical cords (e. g. of microscopes and hot plates) are out of the way of traffic.  Tuck them under the desk.

 

6.   Use hot plates with care:

            -- Remove beakers with hot plates using a suitable protective device  (e. g. insulated

                 gloves or tongs);

            -- Do not let solutions on hot plates boil dry;

            -- Turn off and unplug hot plates after use.

 

7.   Use all chemicals with care:

            -- Read labels carefully before you open bottle;

            -- Do not return unused reagents to the bottle;

            -- Dispose of waste in proper container;

            -- Dispose of biohazardous materials in autoclavable BIOHAZARD BAG;

            -- AVOID GETTING ANY CHEMICAL ON YOUR SKIN OR  CLOTHING;

            -- AVOID BREATHING ANY CHEMICAL FUMES;

            -- if chemicals get into your eyes or on your skin, FLUSH THE AFFECTED AREAS WITH  COPIOUS AMOUNTS

                OF COLD WATER IMMEDIATELY

 

8.   Report major chemical spills to the instructor immediately. 

            -- Do not attempt to clean up such spills yourself  (NOTE:  This includes broken thermometers, which  contain toxic

                mercury).

 

9.   Report immediately all personal injuries to the instructor.

 

10. Be sure to sign the lab safety sheet and return the signed lab copy to the lab instructor.

 

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