SPECTROSCOPIC TECHNIQUES FOR MEDICAL DIAGNOSTIC COURSE March 12 - August 21, 2024

 

Syllabus

Course Schedule

 

Course introduction

• Project management
• Optical design
• Mechanical design
• Electronic design
• Software design
• Product development
• Medical regulations

Fluorescence Spectroscopy

Fluorescence and Phosphorescence Phenomena

• Introduction to luminescence phenomena: fluorescence and phosphorescence
• Distinction between spontaneous emission and non-radiative decay
• Quantum mechanical basis of electronic transitions leading to luminescence
• Fluorescent and phosphorescent molecules in various environments

Jablonski Diagram and Energy Transitions

• Overview of the Jablonski diagram and its role in fluorescence processes
• Singlet and triplet states in the context of electronic energy transitions
• Internal conversion, vibrational relaxation, and intersystem crossing
• Understanding the role of spin and molecular orbitals in energy transitions

Fluorescence Instrumentation and Detectors

• Components of a fluorescence spectrometer and their functions
• Light sources, excitation monochromators, and emission monochromators
• Fluorescence detectors and signal processing
• Considerations for instrument optimization and calibration

Quenching and Factors Affecting Fluorescence Intensity

• Mechanisms of fluorescence quenching
• Common quenching agents and their impact on fluorescence
• Factors influencing fluorescence intensity, such as temperature and solvent effects
• Strategies for mitigating quenching in experimental setups

Time-Resolved Fluorescence Spectroscopy

• Principles of time-resolved fluorescence spectroscopy
• Instrumentation for time-resolved measurements
• Applications of time-resolved fluorescence in probing dynamic processes
• Fluorescence lifetime analysis and its significance

Applications in Medicine

• Medical applications, including diagnostics and imaging
• Exploration of current research trends and advancements in fluorescence applications

Raman Spectroscopy

Scattering of Light and the Raman Effect

• Introduction to light scattering phenomena
• Overview of elastic (Rayleigh) and inelastic (Raman) scattering
• Raman effect: Concept and historical development

Rayleigh and Raman Scattering Processes

• Detailed exploration of Rayleigh scattering
• Energy changes in Raman scattering processes
• Differences between Stokes and anti-Stokes Raman scattering
• Selection rules governing Raman transitions

Raman Instrumentation and Laser Sources

• Components of a Raman spectrometer and their functions
• Types of lasers used in Raman spectroscopy (e.g., diode lasers, Nd:YAG lasers)
• Importance of laser wavelength selection
• Enhancements and advancements in Raman instrumentation

Stokes and Anti-Stokes Raman Scattering

• Understanding Stokes and anti-Stokes Raman scattering
• Factors influencing the intensity of Stokes and anti-Stokes lines
• Practical considerations in Raman spectroscopy for optimizing signal-to-noise ratio
• Applications and significance of Stokes and anti-Stokes shifts

 Interpretation of Raman Spectra

• Fundamentals of Raman spectral interpretation
• Vibrational modes and peak assignments
• Common spectral features and their significance

Raman spectroscopy techniques

• Raman microscopy and imaging
• SORS
• Enhance Raman spectroscopy
• SERS
• TERS
• WERS
• SESORS
• Coherent Raman Scattering
• SRS
• CARS

IR Spectroscopy

Introduction to Infrared Radiation

• Overview of the electromagnetic spectrum, emphasizing the infrared region
• Understanding the molecular basis of infrared absorption
• Energy levels associated with molecular vibrations
• Differentiating between near-infrared (NIR), mid-infrared (MIR), and far-infrared (FIR) regions

Vibrational Modes and Molecular Vibrations

• In-depth exploration of vibrational modes in diatomic and polyatomic molecules
• Relationship between molecular vibrations and infrared absorption
• Symmetric and asymmetric stretching, bending, and rocking vibrations
• Normal modes and selection rules in vibrational spectroscopy

Basics of Infrared Spectrometers

• Components of an infrared spectrometer and their functions
• Dispersive vs. Fourier-transform infrared (FT-IR) spectrometers
• Source, sample compartment, beam splitter, and detector in FT-IR instruments
• Practical considerations and optimization of instrument parameters

Functional Group Analysis and Identification

• Identification of characteristic infrared absorptions for common functional groups
• Correlation between molecular structure and IR spectra
• Spectral fingerprinting for qualitative analysis
• Utilizing IR databases for functional group identification

Interpretation of IR Spectra

• Strategies for spectral interpretation and peak assignment
• Factors influencing peak intensity and shape
• Isotopic effects on IR spectra
• Advanced techniques for spectral deconvolution and resolution enhancement

UV-Vis Spectroscopy

Basic Principles of UV-Vis Spectroscopy

• Introduction to electromagnetic radiation and the UV-Vis region
• Understanding the electronic transitions responsible for UV-Vis absorption
• Concepts of energy levels and transitions in atoms and molecules
• Absorption spectra and their correlation with molecular structure

Interaction of Light with Matter

• In-depth exploration of the interaction between light and matter
• Overview of electronic and vibrational transitions
• Absorption and transmission of light through different materials
• Factors influencing the absorption spectrum

Beer-Lambert Law and Its Applications

• Derivation and explanation of the Beer-Lambert Law
• Relationship between absorbance, concentration, and path length
• Limitations and considerations in applying the Beer-Lambert Law
• Practical applications in quantitative analysis

Instrumentation and Components of a UV-Vis Spectrophotometer

• Components of a UV-Vis spectrophotometer and their functions
• Single-beam vs. double-beam spectrophotometers
• Light sources, monochromators, and detectors
• Calibration and maintenance of UV-Vis instruments

Quantitative Analysis Using UV-Vis Spectroscopy

• Method development for quantitative analysis
• Calibration curves and standard addition methods
• Precision and accuracy in UV-Vis measurements
• Troubleshooting common issues in quantitative analysis

Course Materials: 📚 Required Texts:

1. Handbook of Fluorescence Spectroscopy and Imaging: M. Sauer, J. Hofkens, J. Enderlein, Wiley‐VCH Verlag GmbH & Co. KGaA, 2011.
2. Handbook of Raman Spectroscopy: I. R. Lewis and H. G. M. Edwards, Dekker, 2001.
3. Handbook of Spectroscopy: G. Gauglitz, T. Vo-Dinh, Wiley‐VCH Verlag GmbH & Co. KGaA, 2003.
4. Principles of Instrumental Analysis: D. A. Skoog, F. J. Holler & T. A. Nieman, Cengage Learning, (7th ed), 2018.
5. Project’s schedule

 

INAOE - AGC 2024