We have previously shown that the rhodacyanine dye, Rhodac, exhibits a potent photocytotoxic activity in HeLa cells. In this study several aspects of the photobiological activity of Rhodac were further examined. Rhodac displayed no selective cytotoxicity toward several malignant cell lines after photosensitization (3.6 J/cm²), although HeLa cells were found to be the most sensitive. Interestingly, MCF-7/Adr cells, a multidrug-resistant subline, were less sensitive to the antiproliferative effect of photoactivated Rhodac. The subcellular localization, as revealed by confocal laser microscopy, demonstrated that the dye was mainly concentrated in the cytosolic membranes of the perinuclear region. The Rhodac-induced inhibition of HeLa cell proliferation after light exposure was found to be strictly oxygen dependent. In addition, photoactivated Rhodac induced poly(adenosine 5′ diphosphate-ribose)polymerase cleavage, caspase-3 activation and apoptosis in HeLa cells. In the current work it was further demonstrated that Rhodac binds specifically to high-density lipoproteins and low-density lipoproteins, while no binding was observed to very low-density and heavy proteins. To sum up, our results show that Rhodac is an interesting and potent photosensitizer. Further in vivo experiments are required to elucidate whether the lipoprotein binding leads to a selective uptake of Rhodac in tumor cells and to address its efficacy in photodynamic therapy.