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About 30 students will acquire basic and advanced knowledge on optical microscopy, with an emphasis on fluorescence microscopy. Four two-hour technology lectures will guide the students through the field of optical microscopy, covering a large array of topics ranging from fundamental principles of image formation and fluorescence to modern techniques such as superresolution microscopy (see detailed program). Three one-hour applications lectures from different cell biologists/neuroscientists will demonstrate how these techniques can be employed to solve important biological problems. In addition, four afternoons will be devoted to hands-on practical work, during which students can directly see modern microscopy at work at state-of-the-art facilities (featuring spinning-disk confocal microscopy, TIRFM, single-molecule fluorescence, photo-ablation, superresolution microscopy, STED) and will be trained in quantitative image analysis.
This part will be complemented by training in reading and interpretation of scientific literature, illustrating employment of microscopic imaging techniques. The precise organization of the practical part will strongly depend on the number of participating students.
This one week is meant to be dedicated to this course and students are encouraged to work together as a group and support each other.
Mandatory for students in Master’s programme: NO.
Optional for students in other Master’s programmes GS-LS: CSnD. Neuroscience and Cognition, especially the tracks Experimental and Clinical Neuroscience and any other master programme with relevant basal courses.
The exam (1 hour)
There will be an exam on the material covered in the lectures (50% of the final mark).
In addition, each technology lecture will be preceded by a short examination to test understanding of the basic concepts of the previous lecture. Questions and answers will be directly discussed after the test to support deeper understanding. The average score with contribute 30% of the final mark. The average score should be above a 5 to gain access to the final examination.
Grades
20% of the grade will be based on the short examinations, 50% on the final exam, and 30% on participation and presentations.
Topic covered in the technology lectures
Zooming in on microscopes - basics of light microscopy
- Light and matter
- Geometrical optics
- Image formation and contrast generation
- Koehler illumination, outline of a microscope
- Objectives, numerical aperture and resolution
- Aberrations
On windows and wine
- Fluorescence microscopy
- Basics of fluorescence
- Outline of a fluorescence microscope
- Special fluorescent probes
- Confocal microscopy
- TIRF microscopy
- Multiphoton microscopy
- Light-sheet microscopy
Light at work
- Active-illumination techniques
- Live-cell microscopy
- Active illumination microscopy (FRAP, FLIP, Photoactivation/ablation)
- FRET microscopy (FLIM)
Breaking the barriers
- Supersensitive and superresolved microscopy
- Single-molecules fluorescence microscopy
- Superresolution techniques (PALM, STORM, STED)
- Basics of (EM)CCD and CMOS technology
- Understanding photon counts, noise and grey values
- Comparison of different cameras
- Image processing
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