Biology 132 - Electron Microscopy Lab

      Spring 2004

 

Lisa M. Baird

Lab: MW 14:20-17:20 ST147

Office: ST481, X4073

E-mail: baird@sandiego.edu

Office hours: M-F 11-12

 

Grading:

                        EM technique and completion of objectives on time.............125

                        Project Presentation...............…………………………………..25

                        Project Paper/Microscopy..........………………………………150

 

            Grading will be done on a straight scale; 90-100 = A, 89-80 = B etc. Please note other grading particulars on the syllabus. The Academic Integrity Policy as outlined in the 2002-2004 Bulletin is strictly enforced. If you are uncertain about the policy – particularly as it relates to plagerism - please ask.

 

            Because of the lab intensive nature of the course, you will have a project paper in lieu of a final exam. Your project paper is due the first day of finals on May 13, 2004 by noon. This paper must be typed, illustrated with at least five TEM micrographs from five different fields of tissue. Micrographs should be taken at least two different magnifications (it is not necessary to have two magnifications of each micrograph, but you should show that you can operate the microscope at magnifications above 3000X) and negatives of all micrographs used included in the paper. You should include 5 SEM micrographs from at least 2 different fields of tissue, taken at (at least) 2 different magnifications. Each figure used should include the magnification of the print and a legend describing to the reader what is illustrated. Late papers will lose one letter grade prior to reading unless an extension is granted in advance.

            The text of the paper must be organized and presented in the format of a scientific paper. You need to use at least two primary references in the body of your paper. If you have any questions about this assignment, please ask.

Attendance is expected at all scheduled class and lab meetings. Please note: you should expect to spend at least 5-6 self-scheduled lab hours per week working on your project and completing the lab objectives for the week. Do not procrastinate on your projects!

 

Remember that facilities are limited. All of you can not use a given instrument at one time, nor can I help all of you at once. If you wait until the end of the semester to prepare your project, your grade will suffer. If you are ill or have a death in the family please notify me or the Biology secretary (X4729) as soon as possible. Incompletes will only be given in response to extraordinary circumstance.


 

              Biology 132 - Electron Microscopy

               Laboratory Syllabus and Objectives

Spring 2004

 

Listed below are the lab assignments for each week. The objectives listed are each worth 10 points each and should be completed and submitted to me by the date noted. Failure to complete an objective on time will result in a 10% loss of points for every day (or portion thereof) that the objective is incomplete. If three (or more) objectives are late, your grade will be lowered by one letter. In addition to the objectives, you will be graded on your mastery (through practice!) of EM techniques.

 

Week of:       Lab demonstration/Objective for the week

 

Jan 26            Introduction to the laboratory/discussion of projects

Objective: By Feb 9th identify project idea, outline it briefly and submit it to me (in writing)

 

Feb 2              Principles of fixation, plastics and embedding techniques.

Objective: Select fixation method appropriate to your tissue, obtain necessary specimens and begin fixation by Feb 16. Be sure to have objective checked off when you begin your fixation.

 

Feb 9              Trimming blocks and making glass knives

Objective: By Feb 23rd  tissue should be dehydrated to 70%, begin embedding. Tissue should be in plastic and polymerized by Mar 1st

 

Feb 16            Microtoming I

Objective: Familiarize yourself with the parts of the ultramicrotome. Face off one blank block and have this checked by instructor by Feb 25th.

 

Feb 23            Microtoming II

Objective: Try sectioning on blank blocks, then begin to section your own material. Have your progress noted by instructor.

 

Mar 1              Microtoming III

Objective: Turn in 2 stained slides with your own monitor sections by Mar 17th         

 

Mar 8              Microscope (TEM) I

Objective: Continue sectioning. Familiarize yourself with the basic

controls of the EM

 

March 15-19 - Spring Break

 

 

 

Mar 22            Microscope (TEM) II /Photography

Objective: Practice microscope alignment, observe your own grids. By Mar 24th  all students should be  checked out to operate the EM. Make an appointment  for this check prior to the 24th.

 

Mar 29            SEM tissue preparation/operation of the SEM

Objective: Keep practicing on the microscope. Make an appointment with instructor to assess your progress

 

Apr 5               Darkroom techniques/photo preparation

                        Objective: By Apr19th submit two of your best negatives

 

Apr 12            No demonstration

                        Objective: By Apr 28th submit two prints from any two of your best                               negatives, please include the negative

 

Apr 19                        Group meeting to assess progress

Objective: Bring work completed to date to the lab meeting. With the instructor and your colleagues assess your negatives and progress.

 

Apr 26            No scheduled lab meeting. Time should be used to work on your project and presentation

 

May 3              Oral Project Presentations during scheduled lab period

 

May  13, 2004 Final Paper due by noon.