Fluorescence spectroscopy in biology and medicine

Lecturer:

Tutor:

Supervisor:

Department of Natural Sciences

Synopsis:

The course introduces students to fluorescence spectroscopy and microscopy from its basic physical principles, experimental techniques and working principles of common instrumentation to typical biomedical applications of fluorescence illustrated on selected studies published in literature. Apart from basic methods of fluorescence spectroscopy, new trends in fluorescence microscopy or photodynamic therapy (an example of use of fluorescence phenomena in clinical praxis) will be presented.

Requirements:

Multiple choice test. In case of an ambiguous result followed by a question to discuss.

Syllabus of lectures:

1.Basic principles of optical spectroscopy, absorption of light, electronic transitions

2.Basic principles of fluorescence, Jabloński diagram, fluorescence and phosphorescence

3.Fluorescence spectra and quantum yield

4.Fluorescence lifetime and methods of its measurement

5.Quenching of fluorescence, static and dynamic quenching

6.Polarisation of fluorescence, time-resolved and steady-state fluorescence anisotropy

7.The influence of environment on fluorescence spectra and its use in study of biomolecules

8.Foerster resonance enrgy transfer and excimer fluorescence

9.Protein fluorescence, green fluorescent protein and its mutants

10.Basic principles of fluorescence microscopy

11.Resolution power of fluorescence microscopy and methods of its enhancement

12.Fluorescence correlation spectroscopy and its applications in biological research

13.Photodynamic therapy

14.Recapitulation, test

Syllabus of tutorials:

Study Objective:

1) Understanding principles of the phenomenon of fluorescence and the relationship between its characteristics (spectra, lifetime, polarisation, ...) and the properties of fluorescent molecules and their environment.

2) To learn about experimental methods and instrumentation for characterisation of fluorescence including the state-of-the-art microscopy techniques.

3) Introduction to typical applications of fluorescence spectroscopy and microscopy in biological research and the use of fluorescence phenomena in clinical praxis (photodynamic therapy)

Study materials:

[1]Lakowicz J.R., Principles of Fluorescence Spectroscopy, 3rd Edition, Springer 2006

[2]Prosser V. et al. Experimentální metody biofyziky, Academia 1989

[3]Lecture notes available at course webpage containing links to other freely accessible web sources

Note:

Further information:

No time-table has been prepared for this course

The course is a part of the following study plans:

• Navazující magisterský studijní obor Přístroje a metody pro biomedicínu (compulsory elective course)