3 novel water-soluble indocyanine dyes (em. 690 nm) were designed, synthesized and characterized by nuclear magnetic resonance (NMR) for fluorescent labeling of biomolecules. Spectral properties of the prepared dyes in different solvents were tested. In order to investigate the application of fluorescent dyes in the field of biomedical analysis, the absorption and emission spectra of the dyes in the surfactants as CTAB, SDS were measured, the photostability of dyes were measured by cyclic voltammetry (CV). The application of the new dyes in cellular imaging has been demonstrated through direct conjugating of the dyes with living renal carcinoma cells, then imaging was visualized by near-infrared fluorescence microscopy. The molar extinction coefficient and the fluorescence quantum yield of the dyes were calculated.