Advanced fluorescence techniques
FCH-DA_PFTAcad. year: 2020/2021
The course is designed as a practically focused seminar, which will enable students to acquaint themselves with advanced fluorescence techniques on a suitable model system. Theoretical and practical lessons will alternate within the seminar. The theoretical parts will focus on specific aspects of preparation, measurement and interpretation of the fluorescence techniques and will build on the self-study of the fundamental fundamentals of the given techniques. The practical part includes the preparation of the model system itself (associative colloid system), measurement and evaluation. Within the subject, 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. Together with the introductory lesson of these 6 themes with the theoretical and practical part will create the framework of the semester. At the end of the semester, the basics will be created to create a short monograph on the behavior of the selected system from the perspective of advanced fluorescence techniques.
Learning outcomes of the course unit
The output of the study is practical skills in advanced experimental methods of fluorescence spectroscopy. Students will learn the basics of techniques used in advanced research, will learn the basics of interpretation of such data and will verify this knowledge on a simple system.
Předpokládájí se znalosti z oboru optických spektroskopií a základní znalosti o jevu fluorescence.
Recommended optional programme components
Recommended or required reading
Lakowicz, J.R. Principles of Fluorescence 3rd ed., Springer, Baltimore, 2006 (EN)
Valeur, B.; Berberan-Santos, M.N. Molecular Fluorescence: Principles and Applications, Wiley‐VCH Verlag GmbH & Co. KGaA, 2012 (EN)
Planned learning activities and teaching methods
The course is taught as a set of theoretical and practical seminars.
Assesment methods and criteria linked to learning outcomes
Study results are verified pursuant to Article 34, Paragraph 7 of Study and Examination Rules of BUT. Students are given a credit for submitting protocols for each technique they studied. Protocols are processed in the following format: principles of the technique, the aim and exact methods used for measurement, collected data, their evaluation and interpretation, assessment of the impact of the technique on biophysical practice.
Language of instruction
1) Time resolved emission spectroscopy
2) Time resolved emission spectroscopy - practice
4) Anisotropy - Practicum
5) Resonant energy transfer
6) Resonant energy transfer - practice
7) Fluorescence quenching
8) Fluorescence quenching - practice
9) Fluorescence correlation spectroscopy
10 Fluorescence Correlation Spectroscopy - Practice
11) Two-color fluorescence cross-correlation spectroscopy
12) Two-color fluorescence cross-correlation spectroscopy - practice
13) Presentation of the results of individual techniques
The aim of this course is to introduce and use practical examples of fluorescence techniques that can be used in various disciplines ranging from material sciences to biochemical applications.
Specification of controlled education, way of implementation and compensation for absences
To attend the course, participation in theoretical and practical lessons is required.
eLearning: currently opened course