Radiation phenomena allow us to experience amazing things in life. The blue sky, the sunset, or the rainbow during a soft rain are all result of interactions between light and matter.
In Life sciences, spectroscopy techniques allow us to quantify, visualize and monitor in vivo how our system behaves. Among the possible tools, Fluorescence spectroscopy is one of the most widely used. It can be applied for quantitative colorimetric analysis and/or to label and detect organelles in a virus, bacteria, eukaryotic cell, tissue or even a living organism, among other possibilities. To reach these possibilities, the discovery of fluorescence proteins, as well has the development of bright artificial probes and chemical reactions that would allow site-specific labelling of our desired target, have contributed extensively.
This course will address the fundamentals of Fluorescence spectroscopy, explore the various applications (quantitative and qualitative) that it may be used for and allow the students to get familiar with several techniques that use Fluorescence spectroscopy. It will allow them to pose their questions and address which technique and approach would be the best option to answer their biological question.
• Principles of Fluorescence spectroscopy (Steady state and life-time)
• Sources of Fluorescence in life sciences (natural fluorescence, fluorescence proteins, artificial probes)
• Fluorescence spectroscopy in life sciences (Toxicity, SNR, multicolor)
• Quantitative techniques using Fluorescence spectroscopy
– Partititon, Quenching, FRET
– Co-locallization, FLIM
– Live imaging and quantitative kinetics
– ICS, STICS
– Single molecule techniques (FCS, FCCS and RICS)
– Photoactivation techniques – Superresolution techniques (PALM/STORM)
– Flow Cytometry and kinetics