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Instructors: Dr. Udo Birk; Prof. Dr. Christoph Cremer; Dr. Sandra Ritz
Event type:
Lecture
Displayed in timetable as:
Advanced Microscopy
Hours per week:
2
Credits:
2,0
Language of instruction:
Englisch
Min. | Max. participants:
- | -
Requirements / organisational issues:
Language: English
Lecturers: PD U. Birk, Prof. C. Cremer, Dr. M. Gelléri, Dr. I. Lieberwirth, Dr. S. Ritz
The course will be made available at HTW Chur via Video Conferencing.
Begin 16.04.2019
End 09.07.2019
Main idea:
Resolving structural features at the molecular scale is absolutely essential for harnessing the power of nanotechnology and exploring the unique hierarchical structure of complex biological systems. In this course, we will present the basic concepts of microscopy up to very recent developments in optical and electron microscopy facilitating true molecular resolution. Students will be exposed to cutting edge approaches to molecular resolution in linear and nonlinear microscopy, which are uniquely available in Mainz. Students will be given laboratory tours to see the details of these advanced microscopy systems. This course will be taught at the post-baccalaureate level, basic knowledge in optics is of advantage.
Contents:
16. April: Introduction, Course Organization (Marton Gelleri); Lab Tour: Advanced Microscopy @ IMB Mainz
23. April: NO LECTURES (Homework, Portfolio)
30. April: Physical Basics of Microscopy I (Udo Birk)
- Light, color, contrast
- Pinhole camera
- Optical sensors
- Objectives and optical aberration
7. May: Physical Basics of Microscopy II (Udo Birk)
- Fourier optics
- Optical resolution, pixel resolution and spatial sampling
- Point spread function
- illumination
14. May: Methods of Laser Scanning Microscopy (Microbeam, Confocal Laser Scanning Microscopy (Christoph Cremer):
- Focusing of laser light using high aperture objective lenses
- UV-Microirradiation & some applications
- From Laser microirradiation to confocal laser scanning fluorescence microscopy (CLSM)
- Present CLSM methodology; some biomedical applications
21. May: Focused Nanoscopy (4pi-, STED, MINFLUX) (Christoph Cremer):
- First ideas to overcome the conventional diffraction limit by lens free 4pi microscopy based on holographic focusing;
- Realization of 4Pi microscopy using conventional objective lenses; some applications
- „Breaking the Abbe limit“ using Stimulated Emission Depletion
- Experimental realization of STED microscopy based on objective lenses
- From STED microscopy to 1 nm optical resolution (MINFLUX)
- Towards Lens Free STED/MINFLUX Microscopy
28. May: Fluorescence labeling and photophysics of fluorescent labels (Sandra Ritz)
- review of fluorescence, photophysics, photobleaching
- fluorescence labeling
- Fluorescence recovery after photobleaching (FRAP) for the study of diffusion, transport and metabolic reactions
4. June: Förster Resonance Energy Transfer (FRET) Microscopy; Fluorescence Correlation Spectroscopy (FCS) (Márton Gelléri)
- FRET, Donor quenching, Sensitized Emission, Acceptor Photobleaching, Fluorescence Lifetime Imaging Microscopy (FLIM), Anisotropy Imaging
- Fluorescence Correlation Spectroscopy (FCS)
11. June: NO LECTURES (Homework, Portfolio)
18. June: Single Molecule Localization Microscopy (SMLM), Total Internal Reflection Microscopy (TIRF) (Márton Gelléri)
- Super-resolution imaging by SMLM, localization precision, optical resolution
- Single particle tracking
- total internal reflection, evanescent field, polarization, instrumental approach (prism-type vs objective type TIRF)
25. June: Electron Microscopy I (Ingo Lieberwirth)
- Interaction of electrons with matter, elastic and inelastic scattering
- crystal structure, the reciprocal space, diffraction
- principal components of an electron microscope
2. July: Electron Microscopy II (Ingo Lieberwirth)
- transmission electron microscope, imaging modi, specimen preparation, beam damage
- scanning electron microscope, contrast formation, secondary and backscatter electrons, sample preparation
9. July: Test
subject to modifications
Recommended reading list:
U. Birk, Super-Resolution Microscopy, Wiley
M. Born, E. Wolf: Principles of Optics, Cambridge University Press
R. Gonzales, R. Woods, S. Eddins: Digital Image Processing Using MATLAB, McGraw Hill
M. Gu: Advanced Optical Imaging Theory, Springer
E. Hecht: Optics, Addison-Wesley
E. Saleh, M. Teich: Fundamentals of Photonics, Wiley
F. Träger (ed.): Springer Handbook of Lasers and Optics, Springer
Additional information:
Course webpage: http://www.staff.uni-mainz.de/birku/teach/2019Q1_microscopy.html
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