Temperature affects all biochemical processes. Adaptation to ambient temperature is a key feature that has enabled the survival and development of living beings. Temperature in the cell and its surroundings is a fundamental and extremely important physical parameter for cellular systems and their functions such as cell division, diffusion, metabolism, differentiation, gene expression and enzyme activity. Observing the effects of drugs, disease, aging and other physiological changes in the cell through temperature changes could be monitored in real time using fluorescence.
The phenomenon of fluorescence is the emission of light from a substance, which occurs when a substance is irradiated with light of shorter wavelength, the substance absorbs light, which causes an electron to go into an excited state. Upon returning to the ground state, the compound then emits a photon of a higher wavelength than the light used for irradiation. Compounds that are capable of fluorescence are called fluorophores. The fluorescence intensity of some fluorophores is temperature-dependent and they are a good starting point for the development of probes for measuring temperature at the microscopic level (e. g. in organelles of cells).
In this master thesis, we synthesized various fluorophores which fluorescence depends on the ambient temperature. We designed fluorophores from the family of anthracenes, xanthenes and coumarins, which were also successfully synthesized. The aim was to make water-soluble probes based on different basic skeletons in order to obtain probes with different fluorescence response to temperature. We have introduced an alkine or azide group to these fluorophores, which allow for easy “click” chemistry. After successful synthesis, the excitation and emission spectra of the selected probes were measured. On their basis we were able to decide on the appropriate compounds for further investigation. We then measured the emission spectra of selected compounds as a function of temperature and combined the corresponding pairs with a click reaction. For further work, we propose the synthesis of the same basic skeletons with other substituents to reduce lipophilicity, improve stability and water solubility.
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