1. The fluorescence in analysis of a micelle formation Created by: Jiří Běťák

2. The aim of a lecture - can we describe a process of micellization in the solution of a biopolymer ? Explain: How … - can we use a method of Fluorescence sprectroscopy ? - can we define the Critical micelle concentration ?

3. Vocabulary lesson Probe - sonda highlight - zvýraznit visualise- zviditelnit increase - zvýšení attache - navázat radiation - záření associate – sdružovat se tend to – mít tendenci influence - ovlivňovat volume - objem surface - povrch decrease - snížení

4. The principle of fluorescence Jablonski diagram

5. The fluorescent probes Fluorescent probes are used to visualize investigated molecules that do not emit any visible radiation. The fluorescent probe is attached on a molecules and highlight them. Examples of probes: Pyrene Perylene Antraquinone

6. Characteristics of Perylene Absorption Emission Excitation wavelength: 440 nm Emission wavelength: 450 nm

7. Fluorescence intensity of perylen The more polar system, the lower intenzity. Increace of polarity = decrease of intenzity

8. Process of micellization In the solution particles of a biopolymer consisting of polar and non- polar sides tend to associate influenced by electrostatics interactions. At first they tend to create a monomolecular layer on a surface. When the surface is fully saturated the new particles start to associate into a micelles. The concentration of a biopolymer that fully saturates the surface is called Critical micelle concentration CMC

9. Micellization process of biopolymers Polar Non-polar Particles of a bipolymer – lipides, fatty acids, etc. Non-polarPolar CMC Probe

10. A micelle marking Fluorescent probes that are added into the solution of agregating molecules are freely dispersed in the volume. Their intensity of fluorescence is constant. When the concentration of a biopolymer reaches the CMC, it starts to create a micelles. The probe is electrostatically attracted to the non-polar center of a micelle. The fluorescent probe looses its emitting intensity.

11. The final determination of CMC CMC

12. Thank You kindly for your attention That´s the end…