Course

Unit 1

Emission Spectra, Raman Peaks, Excitation Spectra, Delayed Fluorescence, Scanning Methods, Applications of Polarization/Anisotropy, Protein Oligomerization, Protein/Ligand Interactions, Proteolytic Processing, Fluorescence Polarization Immunoassay, Membrane Fluidity, Effect of Scattering, Rayleigh Scatter and Rayleigh Ghosts, Depolarization via FRET

Unit 2

Excited State Lifetimes, Time Domain, Frequency Domain, Magic Angles, Anisotropic Decay, Dynamic Polarization, Determination of Quantum Yields, Quenching, Collisional or Dynamic Quenching, Static Quenching, Popular Quenchers, Quenching and Membrane Systems

Unit 3

Fluorophores, Intrinsic Probes, Extrinsic Probes, Photostability, Labelling Proteins In Vitro, Non-covalent Probes, Covalent Probes, Photoaffinity Labelling, Membrane Probes, Nucleic Acid Probes, Biosensors, Ion Probes, pH Probes, Molecular Beacons, Voltage-Sensitive Dyes, Quantum Dots, Fluorescent Proteins

LEARNING OBJECTIVES

The syllabus outlines several key learning objectives such as understanding the fundamental principles of fluorescence spectroscopy such as emission and excitation spectra, polarization anisotropy, and quenching mechanisms. Students will learn to apply these concepts to investigate protein interactions, membrane properties, and develop fluorescence-based probes and biosensors. The course also aims to equip students with skills to analyse lifetime measurements, interpret scattering effects, and utilize various fluorescent probes, including quantum dots and fluorescent proteins, in chemical biology research

COURSE OUTCOMES

CO1: Gaining a comprehensive awareness of the principles and techniques of fluorescence spectroscopy, including emission/excitation spectra, polarization, and lifetime measurements.

CO2: Developing the ability to understand various fluorescent probes and sensors, such as protein and membrane probes, quantum dots, and fluorescent proteins, for biological applications.

CO3: Acquiring skills to analyse molecular interactions, protein oligomerization, and membrane dynamics through fluorescence polarization, quenching, and scattering methods.

REFERENCES

1. Principles of Fluorescence Spectroscopy; Joseph Lakowicz

          Springer, New York, 3^rd Edition, 2006.

2. Introduction to Fluorescence; David Jameson

Taylor & Francis, Florida, 1^st Edition, 2014.

3. Essentials of Chemical Biology; Andrew Miller

          John Wiley & Sons, Chichester, 1^st Edition, 2008.