In the first part of the thesis, we generated dye-antibody conjugates by three different strategies which are commonly employed in most research laboratories (amine, sulfhydryl and carbohydrate targeted approaches) so that it could be used for studying membrane protein dynamics. The goal was to obtain conjugates which retained the maximum antigen recognition features and that produced the highest fluorescence intensities. We found that only amine targeted approach could produce higher dye per antibody variants (>3) with a maximum yield of antibody conjugates. Sulfhydryl and carbohydrate targeted approaches are site-specific, however, they led to the generation of low amount of functional dye-antibody conjugates. Therefore, amine targeted approach is the best method of antibody conjugation in comparison with the other two approaches in totality. In the second part of the thesis, our goal was to unravel the topological features of a membrane protein termed cluster of differentiation 1d (CD1d). These proteins are involved in lipid-based antigen presentation. We wanted to demonstrate the relationship of CD1d with peptide antigen presenters i.e. major histocompatibility complex (MHC) proteins on the cell surface. We documented using fluorescent dye-antibody conjugates and biophysical tools that CD1d harbors a close relationship with MHC I, β2-microglobulin (β2m), MHC II and GM1 gangliosides on the plasma membrane of human B cells at resting state. Surprisingly, β2m dependent CD1d constituted only ~15% of the total membrane CD1d proteins. In addition, fluorescence resonance energy transfer (FRET) studies revealed only minimal effect of membrane cholesterol depletion on the association between CD1d and GM1 ganglioside on the cell surface. Instead, CD1d rich regions were highly sensitive to low concentration of Triton X-100. Therefore, CD1d is either located at the periphery of GM1 gangliosides or is enriched in low cholesterol containing detergent sensitive membrane regions of the plasma membrane which could be a distinct raft subtype. In summary, we generated dye-antibody conjugates which were suitable for studying the membrane distribution of CD1d proteins. Subsequently, we also demonstrated that an intricate relationship existed between CD1d, MHC, and lipid species on the surface of human B cells which might be immunologically relevant.