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Tues/Thur 9-10AM, 2304 Tolman Hall
The course will be a detailed overview of the practice of light microscopy as applied to scientific investigation. The emphasis of the course will be on the correct and appropriate use of the light microscope for biological scientists, however students of other disciplines are welcome. We will cover optical microscope theory, microscope components and mechanics, and all modern optical techniques from Brightfield through Deconvolution microscopy. Instruction will consist of lectures, demonstrations, and practice using the light microscopes in the CNR Biological Imaging Facility.
Laboratory: The laboratory will consist of one, 3-hour session per week per student in GPBB Teaching, room 209. Students are to sign up for one lab section:
Tuesday 2-5PM CCN: 70524
Thursday 2-5PM CCN: 71101
Grading: There will be a Midterm written exam and lab practical, plus a written Final Exam. For labs, students will be required to complete one worksheet for each of the techniques covered in lab. Each worksheet is due at the end of that lab. In addition, a project is required for the class. It consists of a written paper. A description of the project will be given toward the middle of the semester.
Syllabus
|
Date
|
Lecture/Lab topic |
|
|
Lecture
|
Topic in Lecture |
Lab topic
|
|
Aug 27
|
Introduction |
|
|
Sep 1
|
Microscope optics: Light and Refraction, Lenses, Imaging and Resolution |
Sample preparation
|
|
3
|
Lens aberrations (spherical, chromatic), The compound microscope |
|
|
8
|
Köhler illumination
|
|
|
10
|
Eyepieces, Inverted Microscope, Stereo Microscope |
|
|
15
|
The Condenser: history, optics & Köhler illumination |
Darkfield
|
|
17
|
Contrast derived from Light Scattering: Darkfield microscopy, Rheinberg Illumination, Oblique Illumination |
|
|
22
|
Polarized light microscopy: Polarized light, Anisotropy, Types of birefringence, Polarized light microscope |
No Lab
|
|
24
|
No Lecture |
No Lab
|
|
29
|
Polarized Light Microscopy
|
|
|
Oct 1
|
Interferometry, Differential Interference Contrast Presentation |
|
|
6
|
Phase Contrast
|
|
|
8
|
|
|
|
13
|
DIC
|
|
|
15
|
Review of transmitted light techniques, Key to determining imaging method, Sample Exam Questions |
|
|
20
|
Midterm Exam (Does not include fluorescence microscopy) |
Lab Exam
|
|
22
|
Lab Exam
|
|
|
27
|
Laser scanning confocal microscopy II |
Fluorescence
|
|
29
|
|
|
|
Nov 3
|
Confocal Microscopy I
|
|
|
5
|
Deconvolution microscopy II, presentation, |
|
|
10
|
High Speed confocal imaging: Spinning Disk & Sweep Scanners, Nipkow disks, Slit scanning confocal microscopes |
Confocal Microscopy II
|
|
12
|
Total Internal Reflection microscopy, TIRF microscope |
|
|
17
|
Advanced fluorescence imaging: FRAP, FRET, FLIM |
Restoration microscopy
|
|
19
|
High Resolution fluorescence microscopy: Structured Illumination |
|
|
24
|
Q & A with Dr. Schichnes |
No Lab
|
|
26, 27
|
HOLIDAY |
No Lab
|
|
Dec 1
|
Digital microscope imaging |
Live-cell microscopy
|
|
3
|
Historical perspective, The Golub Collection |
|
|
8
|
Final Review. Comparison of Advanced Fluorescence Techniques: BF Review, Fluor review, Advanced Review; Project Due |
Optional review
|
|
Dec 15,
8-11AM |
Final Exam |
|
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