Botany 510/BCMB 562
Course Description
Introduction to Electron Microscopy - Transmission Electron Microscopy. This course is designed to give students the necessary skills to prepare and examine biological materials in the transmission electron microscope. Use of the microscope, and ancillary instrumentation, introduction to darkroom techniques and preparation of materials for publication. During the latter part of the semester students will work on their individual research projects.
Syllabus
Week 1. Introduction: Overview of course and requirements.
Source materials. Group lectures on block trimming and knife making.
Introduction to Hitachi H600 electron microscope.
Week 2. Lecture on ultramicrotomy (thin sectioning).
Continuation with microscope training, block trimming and knife making.
Introduction to formavr film making and coating grids.
Demonstration of negative staining.
Week 3. Fixation and embedding of tissue.
Continue with microscope training, block trimming and knike making.
Lecture on fixation and embedding. Introduction to darkroom procedures and printing. Continue with all techniques introduced to date.
Deadline: Turn in quality knives and trimmed blocks.
Week 4. Continue with darkroom procedures.
Introduction to post staining.
Deadline: Turn in negatives of through-focus series of
carbon hole specimen. (Negative magnification x60,000).
Discuss purpose of C-hole exercise.
Week 5. Ultramicrotomy checkout - theory and practice.
Deadline: Evaluation of thin sections.
Demonstration of shadow casting.
Deadline: Turn in prints of C-hole series.
Individual conferences.
Week 6. Continue with above techniques.
Demonstration- calibration of microscope magnification.
Deadline: Turn in 6 negatives of tissue.
Week 7. Demonstration - preparation of micrographs for publication.
Discussion: how to prepare figure legends.
Individual conferences on focus, contrast and print and negative quality.
Introduction to digital image processing.
Deadline: Turn in 5 prints of tissue.
Weeks 8&9. Independent Work.
Week 10. Work. Discuss individual project with faculty.
Week 11. Independent Work.
Week 11. Due by 5 p.m.: Turn in notebook with final electron micrographs, negatives and figure legends.
Begin individual projects on thesis or dissertation material.
SPRING BREAK
Week 12. Individual conferences on notebooks.
Weeks 13&14. Work on individual projects. You must be present during normally scheduled class time. Faculty will be present.
Week 15. Review and evaluate progress on individual projects.
Deadline: Project reports due.
Discuss project reports.
Instructional Personnel:
Dr. John Dunlap
Room 101S&E (4-3590 or 4-6197)
Class meeting time: T-R 9:30-12:30 SERF 101
Requirements:
- Meet deadlines as noted in schedule.
- MINIMUM Notebook Requirements
- 2 negative stain preparations at 2 different microscope magifications.
- 2 shadowcast preparations at 2 different microscope magnifications.
- 8 low magnification prints of tissue. Negative magnification X1000-4000.
- 12 intermediate magnification prints of tissue. Negative magnification X10,000-50,000.
- 4 high magnification prints of tissue. Negative magnification X75,000 & above.
- Note: Prints for all of the above:
- One 8X10 print at each magnification
- All others prints 4X5
- 1 Plate consisting of at least 5 different micrographs. Specifics will be given in class.
Examples of Students Work
