The thesis deals with the study of hydrophobic domains in phase separated hydrogels. Two types of polyelectrolytes (hyaluronan, dextran) and oppositely charged surfactants (carbethopendecinium bromide, sodium dodecylsulphate ) are used. Phase separated hydrogels are formed by electrostatic interactions. The target of the thesis is to study the polarity of environment in phase separated hydrogeles by fluorescence spectroscopy. Fluorescence measurements is performed by using pyrene, Nile red and laurdane as fluorescence probes. All probes are sensitive to the polarity of the environment. The results showed that probes can be expected in the polar environment of hydrogels.