The design and synthesis of new fluorescent dyes with desirable photophysical properties for labelling amino and hydroxyl groups are of considerable interest in bio-analytical research. This research is principally focused on the synthesis, characterization and investigation on the optical properties of dyes based on pyrazoline and furanone skeletons in different media. The feasibility of new dyes was checked by synthesizing its WAS corresponding derivatives. Theoretical calculations were also performed and the experimental values were correlated with the theoretically derived results. Chapter 1 of this thesis presents a concise introduction to the fundamental concepts of fluorescence and importance of fluorescent labelling techniques in present scenario. In this chapter we also highlight some similar studies which utilized the molecular moieties of our interest. Furthermore, the objectives and highlights of this project are also explained in this chapter. Chapter 2 describes the methodology that we adopted through the different phases of this project. Chapter 3 deals with the detailed experimental and theoretical results we achieved in this study. Moreover, we tested the potentiality of one of our new dyes using analytical techniques, HPLC and LC-MS and is reported in Chapter 4. The thesis concludes with a brief Chapter 5, whịch describes the applications of newly developed compounds in various dimensions in account of their excellent photophysical properties. Chapter 6 suggests some of our future directions for more extensive applications of novel dyes we developed in various fields of current interests.