Quantitative and Analytical Light Microscopy

Lecturer(s)

Dr. Y. Belyaev, Dr. G. Csucs, Prof. A. Stemmer

Hour(s) of lecture(s), exercise(s) and credit points

4G (total hours of lectures/exercises)
4CP

Teaching goals

In-depth theoretical and practical knowledge of light microscopy as a means to study life processes on the tissue, cellular, and molecular scale. An explicit objective of this class is to bring together students from engineering and biology and to train them in transdisciplinary information exchange.

Summary and outline

Theory of image formation in the light microscope. Components (optics, cameras) and alignment of a microscope. Aspects of resolution. Super-resolution microscopy (HELM, model-based image analysis). Non-fluorescent contrast modes: bright-field, dark-field, phase contrast, differential interference contrast (DIC). Fluorescent labelling techniques. Epi-fluorescence. Laser Scanning Confocal Microscopy. Multi-photon fluorescence microscopy. Specialized techniques: Fluorescent Speckle Microscopy to measure dynamics of protein assemblies in living cells; Fluorescence Recovery After Photobleaching (FRAP) to measure diffusion processes; Fluorescent Correlation Spectroscopy (FCS); Fluorescence Resonance Energy Transfer (FRET) to measure molecular interactions. Hybrid microscopes.Block course in the first week of the spring semester break. The idea of this block course is to allow students getting hands-on experience in imaging using high-end research microscopes in the Light Microscopy Centre and the lab of Prof. Stemmer. Students are welcome to bring their own samples. Specific experiments on specialized instruments can be pursued upon request.

Literature

Lecture notes