The optical properties (visible-ultraviolet emission and fluorescence) of acridine orange (AO) were compared and investigated to try to discuss the AO adsorption mechanisms when AO interacted with four aqueous dispersions, for example, single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), hybrids of ssDNA and single-walled carbon nanotubes (SWNT-ssDNA), and hybrids of dsDNA and single-walled carbon nanotubes (SWNT-dsDNA). Molecule structure deformation and energy transfer maybe existed when AO attached to DNA chains or emerged into SWNT/DNA suspensions. DsDNA seemingly brought about an obvious structure deformation to AO; delocalized π-bond of AO/SWNT perhaps produce a little absorbance red-shift effect to AO, because of lower electronic transition energy for π-electron or n-electron; the energy transferred from DNA to AO, or from AO to SWNT. Most of AO molecules “prioritized” DNA chains, rather than SWNT surface; although AO molecules adsorbed to DNA chain and SWNT surface at the same time.