Course Program

  1. Light-matter interactions and optical microscopy
    1. Light, color, waves
    2. Geometrical optics
    3. Diffraction and interference in image formation
    4. Microscope design
    5. Diffraction and spatial resolution
    6. Contrast in image formation: bright-field and dark-field microscopy
  1. Fluorescence microscopy
    1. Definitions, empirical approach, quantum-mechanical basis of the fluorescence process, Jablonski diagram
    2. Optical read-outs of fluorescence: intensity, wavelength, lifetime and anisotropy
    3. Excited state reactions and relaxation
    4. Fluorescence quenching
    5. Optical microscopy and fluorescence microscopy
    6. Confocal microscope
    7. Two-photon microscope
    8. Total Internal Reflection Microscope
    9. Binding by fluorescence microscopy: Forster Resonance Energy Transfer
    10. Dynamics by fluorescence microscopy: Fluorescence Recovery After Photobleaching and Fluorescence Correlation Spectroscopy
  1. Super-resolution microscopy
    1. Toraldo’s statement on the physical basis of resolution
    2. Bistability and information
    3. The stochastic approach: PALM, STORM, and SOFI techniques
    4. The deterministic approach: STED and RESOLFT techniques
    5. Structured illumination

Bibliographic references

P. P. Mondal and A. Diaspro "Fundamentals of Fluorescence Microscopy", Springer Science, 2014

M. Muller "Introduction to Confocal Fluroescence Microscopy" Spie Press, 2nd Ed. 2006

ExamOral exam