Light microscopy for biological samples

The following topics are treated: perception, light, lenses, the microscope, brightfield microscopy, darkfield microscopy, phase contrast microscopy, (quantitative) polarised light microscopy, differential interference contrast microscopy, fluorescence microscopy, fluorescence recovery after photobleaching, Förster energy resonance transfer, total internal reflection fluorescence microscopy, laser scanning microscopy, confocal microscopy, spinning disk confocal microscopy, multi-photon excitation microcopy, second harmonic generation microscopy, (dual colour) fluorescence (cross) correlation spectroscopy, fluorescence lifetime imaging microscopy, single molecule localisation microscopy, photo-activated localization microscopy, (rapid) (direct) stochastic optical reconstruction microscopy, stimulated emission depletion microscopy, ground state depletion microscopy (with individual molecule return), live cell imaging, electronic imaging.

The course is meant for students who may have to do light microscopy on biological samples for their internship(s) or MSc-project, and for students who are interested in biomedical applications of optics in general. After completion of this course one will have obtained a thorough background in the possibilities and limitations of various optical-microscopic techniques, in particular the possibilities and limitations related to biological samples.

The student can:

• explain the physical and optical background of the microscopy techniques that are available in the MaTe Laboratory for Cell and Tissue Engineering
• explain the purposes and limitations of all the microscope parts in the microscopes that are available in the MaTe Laboratory for Cell and Tissue Engineering
• judge the microscopy techniques that are available in the MaTe Laboratory for Cell and Tissue Engineering for their benefits and pitfalls, and for fitness for (a given) purpose
• for a given purpose, judge the quality and fitness of microscopy images in literature
• given a specific problem, judge on the most suitable microscopy technique for that problem

You must meet the following requirements

• Completed Final examination Bsc program

• Fundamentals of light microscopy and electronic imaging (ISBN 978-0-471-69214-0)
• Handouts of lectures.
• Extra information handed out during the lectures. Students are expected to find their own way in literature.