BIO340 Animal Physiology

Fall 2019

Laboratory guide

 

 









 

BIOLOGY 340 LABORATORY SCHEDULE

 

WEEK       DATES                            TOPIC                                                                           

         1

August 22

Lab 1: Electrophysiological Instrumentation

         2

August 29

Lab 2: Permeability of the RBC membrane

           Worksheet for Labs 1 & 2  due at start of lab

         3       September 5

Lab 3: Active Transport in the Frog Skin

            Worksheets for Lab 3 due at start of lab

         4       

September 12

Lab 4: CAPs in the Frog Sciatic Nerve

            Worksheet for Lab 4 due at start of lab

         5       

September 19

Discussion: Labs 1-4

                    Questions from Lab 1 due at start of lab

         6      

September 26

Midterm I

         7       

October 3

Lab 5: Sensory Coding by APs in the Cockroach Leg

           Worksheet for Lab 5 due at start of lab

         8      

October 10

Lab 6: Excitation and Contraction of Frog Muscle

           Worksheet for Lab 6 due at start of lab

Oct 11 (by 5 PM) 1 or 2 Data Sheets from Labs 2-4 Due*

         9     

October 17

Lab 7: Microvascular Circulation in the Frog Tongue

            Worksheet for Lab 7 due at start of lab

        10       

October 24

Discussion: Labs 5-7

        11

October 31

Midterm II (Happy Halloween!)

        12       

November 7

Lab 8: Control of Heartbeat in the Frog

            Worksheet for Lab 8 due at start of lab

        13

November 14

Lab 9:  Blood and Urinary Glucose in Humans

             Worksheet for Lab 9 due at start of lab

Nov 15 (by 5 PM) 1 or 2 Data Sheets from Labs 5-7 Due*
        14 November 21

Lab 10: Human Physiological Responses

              Worksheet for Lab 10 due at start of lab

              

November 28

NO LAB - THANKSGIVING BREAK

        15       

December 5

Discussion Labs 8-10

        16

Dec 11 (by 5 PM)

1 or 2 Data Sheets from Labs 8-10 Due*

 

12/16 Mon 8:30 AM

Lab Final following Class Final 

 

* You will turn in 5 Data Sheets – up to 2 sheets each from three sets of labs (2-4, 5-7, 8-10).  Lab 1 has no data sheet.  Data Sheets are due by 5:00 PM on Friday October 11, Friday November 15, and Wednesday December 11, as indicated.  There will be a 5 point grade reduction for each sheet turned in late. No written work will be accepted after 5:00 PM on Thursday December 12.


 

back to top=


 

INTRODUCTION AND OVERVIEW

 

     The laboratory portion of this course is designed to complement the lecture/discussion part of the course.  It will provide you with hands-on experience in testing some of the physiological principles which we will be discussing in class, in vivo (in living organisms), in vitro (in isolated, functioning tissue), and ‘in silico’ (in computer and electronic simulations).  The study of physiology is manipulative in nature, and there is no substitute for direct experimentation.  Contrary to what you might have heard or read, there is also no real substitute for working with real animals. 

 

      This laboratory is not intended to be an exhaustive practicum in physiological methods, but rather to get you comfortable with some basic physiological instruments applied to basic physiological questions, as well as to introduce you to basic physiological reasoning.  The exercises are adaptations of classical physiology experiments, which lead you through a predetermined sequence of procedures.  You will have the opportunity to practice experimental design directly at the end of each laboratory exercise, by testing novel questions of your own. 

 

Completing the laboratory portion of this course will require a good deal of work and a good deal of time.  Most of you are taking this course as preparation for postgraduate training and a professional career.  This course will allow you to acquire a base of knowledge and experience sufficient for you to compete successfully with any undergraduate student in the country, if you are willing to put in the necessary effort.  Consequently, the demands made on you in this course are designed to be in line with the demands made upon physiology students in the best undergraduate institutions. 

 

      As is the case in any laboratory endeavor, the first key to getting the most out of your experience is to prepare before you to come to the lab.  For each of the laboratory exercises you should read through the laboratory guide, and through the assigned readings.  Be sure that you understand the rationale for each experimental approach and for each sequence of experimental steps.  Look over any equations and try to understand how they are applied and how they were derived.  Complete the worksheet to review your understanding of the lab.  Set up your data collection files and review exactly what data you will need to collect to successfully complete the Data Sheet.  Come see the instructor with any questions.  Do this before each lab period starts.

 

      The second key to success in the lab is meticulous record-keeping.  A laboratory notebook is not strictly required, but is strongly recommended.  The easiest way to make such a notebook is to add your laboratory notes into the lab manual.  As you perform each experiment, keep a running record of equipment settings, observations, procedural mistakes and apparent anomalies, numerical results, preliminary calculations and graphs, etc.  Add these notes to the appropriate section of your notebook, along with copies of your worksheets and data sheets.  Keep your PowerLab data on a floppy disk with your laboratory notebook.  If you want, you are free to break up this lab manual and partition it out in your lab notebook.  The important thing is to establish a system that works for you, and then stick to it.  Your notebook will not be collected or graded, but it can provide a simple way to keep your laboratory notes organized.  Who knows?  One day you may want to teach a physiology lab and find yourself frantically searching your notes to figure out what size microammeter you need for the frog skin experiment.

 

      The third key to success is time.  Three hours is actually a very short period in which to complete a physiology exercise.  Frankly, most of these labs work better at four hours.  Getting your equipment set up and your physiological "preparation" prepared will take 1 to 1 1/2 hours, assuming that you have studied the instructions, are reasonably dexterous, and know exactly what you are doing.  You will have to use the remaining time very efficiently in order to finish "on time".  It is not appropriate to leave without finishing a laboratory exercise.  You should, therefore, get used to the idea that many labs will run past the designated ending time.  This will be the case in many of the upper-level biology courses at Wesleyan, and in any graduate laboratory course you take.  Late-running labs are simply one of the costs of being a scientist.  I am prepared to stay until you have finished the laboratory; you should be, too.

 

      Finally, you should be aware that this laboratory involves a good deal of animal dissection.  Read through the labs and make sure that this prospect does not unduly disturb you.  If you are think that you are going to have problems with this aspect of the experiments, please talk to me as soon as possible.

 

back to top=


 

LABORATORY ASSIGNMENTS

 

Laboratory Participation

 

      30 points (7.5% of the laboratory portion of the course grade) is reserved for a lab participation score.  This is a purely subjective grade given you by the instructor.  If you reliably show up for labs, work well with your lab partners, and contribute your fair share, you will get all 30 points.  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 will get less (substantially less) than the full 30 points.  Physiology labs require cooperation and this is my blunt way of making sure that you do just that.

 

Lab Questions

 

      You will be required to hand in a complete set of answers to the questions in the text of the guide to Lab 1.  These answers must be your own, i.e. written in your own words and substantively different from the answers of your classmates.  This is to insure that you have completed this instrumentation lab, and understand the PowerLab system well enough to not be a burden on your lab partners.  This answer set will be worth 10 points (2.5 % of the laboratory portion of the course) and is due at the start of the laboratory discussion period on September 14.

 

      You do not have to turn in answers to the questions imbedded in the remaining labs, but answering them as you go would be a really good idea. Text questions from the remaining labs will serve as the basis for the lab portion of the final exam.

 

Worksheets

 

      Each of the labs 1 through 10 has a short worksheet associated with it.  These worksheets are included at the end of each laboratory guide.  The answers to the questions on each worksheet can be found in the laboratory guide, or in the assigned readings for that lab.  The worksheets are designed to make sure that you have read and grasped the important features of each lab, before you come to the laboratory session.  The questions range from simple to moderately difficult, and you should be easily able to get a nearly perfect score. 

 

      Each of the 10 worksheets is worth up to 10 points (2.5% of the laboratory portion of the course grade), but only the top 8 grades will be recorded.  Each worksheet is due at the start of the lab period. You will not be allowed to start any lab without turning in the appropriate worksheet.

 

Data Sheets

 

      You will turn in your results from labs 1 through 10 in the form of Data Sheets.  Each sheet corresponds to the "meat" of a full scale lab write-up, without any of the "trimmings".  It will consist primarily of properly labeled and annotated graphs, tables, computer printouts, and calculation results based on the data which you have gathered during the experiment.  Specific instructions for what to include in each data sheet are provided at the end of each laboratory guide.

 

      You will turn in a total of five of these data sheets: a maximum of two data sheets from each of three sets of labs (1-3, 4-6, 7-10).  Note that labs 0 and 8 have no data sheets associated with them.  Each of the six data sheets is worth up to 36 points (9% of the laboratory portion of the course grade).  Due dates for these data sheets are indicated in the Laboratory Schedule at the beginning of this manual.  If a data sheet is turned in late, 5 points will be taken off for each week or fraction of a week elapsed since the due date.  No data sheet will be accepted after Wednesday, December 6 at 5:00 PM.

 

Laboratory Examination

 

      A single laboratory final exam will follow the class final exam.  At least half of the lab exam questions will be drawn from the questions included in each laboratory guide and the questions on the worksheets. The remainder of the lab exam questions will deal with either experimental designs or data interpretation related to the laboratory exercises.  The laboratory exam will be worth 100 points (25% of the laboratory portion of the course grade).

 

back to top=


 

LABORATORY GRADING

 

      The laboratory portion of the course will count for 40% of the total course grade.  Points for the laboratory portion of the course will be assigned as follows:

 

Assignment                                                                           Pts.      lab %      course %

Lab 1 Answer Set                                                                 10          2.5%             1%

Worksheets  (8 Worksheets @ 10 points each)                    80         20%               8%

Data Sheets  (5 Data Sheets @ 36 points each)                 180         45%             18%

Laboratory Final Examination                                           100         25%             10%

Laboratory Participation                                                      30          7.5%             3%

TOTAL                                                                             400        100%            40%

  

back to top=


 

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 MacLab A/D hardware box (retrofit as a PowerLab), and PowerLab software.  The computers are named Christa, Judy, Ruth, and Sally (in honor of four women who contributed significantly to scientific exploration).  These computers are here for your use.  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)   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.

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

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

4)   Don’t download any files to the computer.

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

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

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

8)   Report any computer housekeeping or etiquette problems to me as soon as possible.

 

back to top=


 

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. g Ascaris, 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.

 

back to top=