ADVANCED OPTICAL MICROSCOPY FOR LIFE AND MATERIAL SCIENCE
Obiettivi di apprendimento
Offer an overview of the principles of modern light miscroscopies in life and materials science.
Theory of light, light matter interaction, with an emphasis on fluorescence
The Light Microscope: history and evolution, components of a compound microscope
- The optical components of compound microscope and their functions/operations
- The physics of light and glass
- Forming and maintaining optical resolution with these lenses
- Proper(Kohler) bright field illumination
- Principles of sample preparation for light microscopy
- Theory of image formation, diffraction-limited resolution
- Optical aberrations and their correction
- Illumination and image forming techniques:
* Brightfield Optics
* Oblique Illumination Techniques
* Imaging of Phase Objects
* Polarization Optics
* Fluorescence Microscopy
* Probes for fluorescence microscopy
* Confocal Microscopy
* Single molecule microscopy and applications in biology
* FCS and applications in biology
- Labeling and labeling strategies
- DNA sequencing with emphasis on single molecule optical approaches
- Non-linear microscopy: Multi-photon excitation, Second Harmonic Imaging, different Raman microscopy schemes
- New microscopies to improve resolution
- STED, structured light microscopy, PALM/STORM
Case studies in biology, biotechnology, material science,.
• Structural and functional analysis of cellular and subcellular entities at various length scales and with sensitivities down to the single molecule level via optical microscopy
• Introduction to DNA sequencing, an overview of the different technologies. The role of microscopy in DNA sequencing will be explained. Optical mapping as a coarse (compared to sequencing) but useful DNA tool will be outlined.
• Technical principles of electron microscopy (EM) and atomic force microscopy (AFM) will be explained. Salient examples of the use of EM and AFM in a nano-biological context will be explained. The link with optical microscopy and the differences in resolution, dynamics, contrast, will be emphasized.
Original research papers and reviews published on scientific journals
The course consists in a series of lectures covering the fundamentals of advanced light microscopies.
Part of the course will be the reading of a microscopy related paper and presentation discussion on the paper in one of the classes.
A selection of papers will be presented from which students can choose for their presentation.
Modalità verifica apprendimento
Oral. 25 % of the points will be given for the presentation on a paper chosen from a selection.