Course detail

Advanced Fluorescence Spectroscopy

FCH-MC_PFSAcad. year: 2024/2025

Learning outcomes of the course unit The aim of the course is to introduce students to various fluorescence techniques and their application with interpretation of obtained results. Students will be introduced to the phenomenon of fluorescence from the basis of light-matter interaction to super-differentiated microscopic techniques. The following techniques will be discussed: time-resolved emission spectroscopy, anisotropy, resonance energy transfer, fluorescence quenching, fluorescence correlation spectroscopy, two-color fluorescence cross-correlation spectroscopy. Practical examples, their advantages, limitations and methods of interpretation will be presented for each method.

Language of instruction

Czech

Number of ECTS credits

Mode of study

Not applicable.

Guarantor

doc. Ing. Filip Mravec, Ph.D.

Department

Institute of Physical and Applied Chemistry (ÚFSCH)

Entry knowledge

physics, physical chemistry

Rules for evaluation and completion of the course

The acquired knowledge will be verified by an exam, which will have a test and an oral part. The understanding of individual phenomena, knowledge of basic fluorescent techniques and methods of data evaluation will be verified.
Attendance at lectures is optional, recommended.

Aims

Learning outcomes of the course unit The aim of the course is to present basic and advanced information about techniques using the fluorescence phenomenon.
Graduates of this course will understand the basic concepts of fluorescence spectroscopy. They will orientate themselves in stationary, time resolved and microscopic techniques. They will be provided with practical information on how to measure and analyze relevant data.

Study aids

Not applicable.

Prerequisites and corequisites

Not applicable.

Basic literature

Lakowicz, J.R. Principles of Fluorescence 3rd edition. Baltimore, Springer, 2006 (CS)
Valeur B., Berberan-Santos M.N.: Molecular Fluorescence: Principles and Applications. Weinheim, Wiley‐VCH Verlag GmbH & Co. KGaA, 2012 (CS)

Type of course unit

Lecture

26 hod., optionally

Teacher / Lecturer

doc. Ing. Filip Mravec, Ph.D.

Syllabus

- What is light and matter? Historical overview. Description tools.
- Scattering and absorption of radiation - electron, vibrational and rotational transitions. The fate of the excited state. Kinetics. Nonlinear optical phenomena.
- Solvatochromism and other environmental effects.
- Absorption spectrophotometry - uv-vis-ir, spectrophotometers and advanced techniques.
- Fluorescence spectroscopy stationary - fluorimeters, types of spectra, quantum yield
- Time-resolved fluorescence spectroscopy - time vs. time frequency domain, instrumentation and principles
- Microscopy - visible, fluorescent, confocal - principles and instrumentation
- Fluorescence correlation spectroscopy - autocorrelation function, cross-correlation function, instrumentation, measurement principles, models
- "Supermicroscopy" - STED, dSTORM, PALM, PAINT etc. - principles and applications
- The most common mistakes and errors in measurement.

Guided consultation in combined form of studies

26 hod., optionally

Teacher / Lecturer

doc. Ing. Filip Mravec, Ph.D.