This study outlines the synthesis, characterization, optical properties and DFT studies of new donor-π-acceptor fluorophores. The basic design of fluorophores included 1,3-indane dione as new acceptor unit, cyclohexyl fused π-linker and indole units as donor groups. Furthermore, selected fluorophores were modified at meso Cl position with O, S and N bearing moieties. The structure of fluorophores was characterized with ¹H and ¹³C NMR. The fluorophores exhibited strong absorbance and emission in NIR region between 750 nm and 830 nm. The fluorophores with N meso modification showed significant blue shift in absorbance spectra leading to large Stokes shift in optical profiles. Some of the fluorophores showed distinct J-aggregation and H-aggregation which was further studied in selected fluorophores. The DFT studies were used to understand charge transfer character of the fluorophores, and energy levels of frontier HOMO/LUMO orbitals. Finally, in vivo biodistribution studies were analyzed for potential bio targetability of the fluorophores.