415/515 Immunology Principles and Practice

Laboratory Syllabus - Fall 2009

August 24 and 25

Introduction and Overview

Dilution Techniques

August 31 and September 1

Agglutination

Quantitative Precipitation

September 7 and 8

Labor Day Week - No Laboratory Sessions

September 14 and 15

Agglutination

September 21 and 22

Ouchterlony Double Diffusion

Radial Immunodiffusion

September 28 and 29

Immunoelectrophoresis

Immunoblot Analysis

October 5 and 6

Fluorescent Microscopy

Flow Cytometry

Review for Examination 1

October 12 and 13

Examination 1 (Safety ... Flow Cytometry, plus relevant sections of Chapter 6, Kuby Immunology)

Notebook Assessment 1

October 19 and 20

Enzyme-Linked Immunosorbent Assay

TLRs and Cytokine Production

October 26 and 27

Cytokine Assays

November 2 and 3

Cytokine Assays

November 9 and 10

Phagocytosis and Opsonization

November 16 and 17

Bacterial Clearance

November 23 and 24

Antibody Response Analysis

Laboratory Report Due (TLRs and Cytokines)

November 30 and December 1

Hypersensitivity

Review for Examination 2

Student Evaluation

December 7 and 8

Examination 2 (Enzyme-Linked Immunosorbent Assay ... Hypersensitivity, plus relevant sections of Kuby Immunology, Chapter 6)

Notebook Assessment 2

Immunology Laboratory Manual. 2009. John R. Stevenson and Joseph M. Carlin, Department of Microbiology, Miami University, Oxford, OH.

Additional Reading

Antigen-Antibody Interactions (Kuby Immunology, Chapter 6)

Laboratory Instructors

• John R. Stevenson, PhD
  • Course Director
  • Office: 52 Pearson Hall
  • Phone: 529-5427
  • E-mail: StevenJR@muohio.edu
  • Hours: MTWRF 2:00-3:00pm (or by appointment)
• B. Jason Hayes, BS, MS
  • Section A: Monday 8:00-9:50 am
  • Office: 50 Pearson Hall
  • Phone: 529-5440 or 529-7262
  • E-mail: HayesBJ2@MUOhio.edu
  • Hours: By appointment
• Jason P. Clark, BS
  • Section B: Monday 2:10-4:00 pm
  • Office: 64 Pearson Hall
  • Phone: 529-5442 or 529-7263
  • E-mail: ClarkJP2@MUOhio.edu
  • Hours: By appointment
• Ryan F. Relich MT(ASCP)cm
  • Section C: Tuesday 10:00-12:00 am
  • Office: 78 Pearson Hall
  • Phone: 529-5442 or 529-7265
  • E-mail: RelichRF@MUOhio.edu
  • Hours: By appointment

Objectives

The field of immunology has continued to develop at a rapid pace throughout the 20th century and during the early years of the 21st century. Consequently, numerous assays and techniques are available to the professional immunologist. It would not be possible to incorporate all of these in any single course. Therefore, the purpose of this laboratory course is to provide a selective approach to your learning of certain immunologic techniques and assays.

Attendance

Whereas you are expected to participate in every laboratory exercise, only one excused absence will be given to students with valid medical excuses or reasons acceptable to their TA. Each additional absence will result in deduction of ten percentage points from your final percentage.

Laboratory Preparation Information

For best performance of the laboratory exercises and best understanding of the course material, it is important to approach the laboratory exercises in a thoughtful, organized way. This includes reading each exercise and writing a half-page of laboratory preparation informationfor each lab exercise on what you will be doing during class before you come to the laboratory. You must turn in this "lab prep info” at the before class begins as evidence of your advanced preparation. Your lab prep info must contain a description of the experimental approach to be used in the exercise together with a description of how the data will be reported. (Note: some days have more than one lab exercise; therefore, you must write more than one lab prep.) Failure to turn in this information on time will result in deduction of five points from your total course points.

Quizzes

On occasion, your Laboratory Instructor will give you an unannounced quiz on the material he/she thinks you should understand to best perform that day’s exercise(s).

Notebook

Your notebook must be an up-to-date, accurate account of everything you do in this laboratory. It should use the same basic format as your laboratory reports, but is not expected to be as rigorous. This is highly important if you are to get the most out of your laboratory experiences this semester. Your laboratory instructor will assess your notebook twice during the semester to assure that you are developing the record-keeping habits expected of professional microbiologists. Your notebook entries should follow this format:

• Introduction (Lab Prep Info) - What is the purpose of this exercise (What will you learn) and how will you accomplish it?
• Materials and Methods – This section consists of changes in the protocol and/or clarifications of instructions to be written in your notebook.  What did you do and what did you do it with?
• Results - What data did you obtain? (Tables and figures are generally the best approach for recording data; text is needed for further understanding).
• Discussion - What do your results mean?

Laboratory Report

You will generate one laboratory report based on the Antibody Response Exercise. It should be well integrated, and follow this format:

• Introduction - "Set the scene" for the rest of the report
  • Background: Explain any theoretical background necessary for a clear understanding and evaluation of the results of the experiments, including a review of pertinent literature, a general statement of the method used and why it was chosen, and a statement of the principal results of the study.
  • Purpose: State the purpose of the experiment (describing the nature and scope of the problem) and explain what the experiment accomplished
• Materials and Methods - Provide all the technical details of the experiments (sufficient that a competent worker can repeat them)
  • Materials: Describe what materials were used to do each technique, including exact technical specifications such as concentration, quantity and source (or method of preparation) for each item and organism.
  • Methods: Describe, in detail, the methods and procedures by which each technique was done, including precise details about conditions such as temperature, time, centrifugal force, volumes, voltage, current, etc.
• Results - Present the data in an understandable framework, by:
  • Text: Describe experiments in a way that makes the processes and results obtained clear, but resisting the urge to repeat experimental details described in materials and methods.
  • Tables/Graphs: Present data obtained using tables, graphs and/or diagrams as necessary for clarity:
    • Do not repeat data presented in tabular, graphic or diagrammatic form in the text; but …
    • Do point out trends that clarify interpretation of the results; however, don't discuss them yet (save that for the Discussion).
• Discussion - This section should interpret the results and explain their relevance by pointing out and discussing (considering from all reasonable angles) the relationships among the observed facts (your data vs. those of others)
  • Interpretations: Present principles, relationships, or generalizations shown by the results;
  • Exceptions: Pointing out any exceptions or lack of correlation and designating any indeterminate points as being unsettled;
  • Comparisons: Explain how your results and interpretations agree (or disagree) with previously published work;
  • Perspectives: Consider from all reasonable perspectives any theoretical implications and practical applications of your results;
  • Conclusions: Summarize the evidence for each conclusion, then state it as clearly as possible.
• References - This section should describe all sources of information used in designing your experiments, as well as in interpreting your results. Be sure you refer to these references at appropriate places in your report. Use these formats:
  • Journal article: Davis, Norine J., and John R. Stevenson. 2003. Role of neutrophils in decreased resistance of protein-malnourished mice to Candida albicans. Nutr. Res. 23:945-958.
  • Book compiled by an editor:  Stocker, B.A.D., and P.H. Makela. 1986. Genetic determination of bacterial virulence, with special reference to Salmonella, vol. 124, p. 149-167. In D.E. Briles, (ed.), Current Topics in Microbiology and Immunology. Springer-Verlag, New York.
  • Book not compiled by an editor:  Day, R.A. 1979. How to Write and Publish a Scientific Paper. p. 38. ISI Press, Philadelphia, PA.

In preparing your laboratory reports, use appropriate material from your textbook, laboratory manual, or lecture notes as well as other books and articles. You should not, however, simply "regurgitate" any of this material (especially the procedural details), because the object of this report is for you to integrate the information and present it in a clear and thoughtful manner in your own words. These reports must be generated using a computer-based word processor. The Department of Microbiology has a computer laboratory set up for student use. Ask your laboratory instructors when you can use this facility to complete your report.

Plagiarism and other academic misconduct. Academic honesty is expected and required. Be sure your words are your own, on both exams and reports. All cases of plagiarism or other academic misconduct will be referred to the Office of Judicial Affairs. Miami University’s policy on academic misconduct can be found in your student handbook and on-line at: http://www.miami.muohio.edu/documents_and_policies/handbook/

Evaluation

Your grade in Immunology Laboratory will be determined by your performance on examinations, quizzes, notebook assessments, and lab report. The following weightings will be used in determining your grade:

• Examinations:
  • Exam 1 = 25%
  • Exam 2 = 35%
• Quizzes (10%)
• Notebook assessments:
  • Assessment 1 = 10%
  • Assessment 2 = 10%
• Lab report (10%)

Your Laboratory Instructor will determine your scores on your examinations, quizzes, notebook assessments and laboratory report. All reasonable efforts will be made to assure equality of grading among sections. 

Cheating on any aspect of this course may make you subject to immediate dismissal from the class with a grade of F.