Four rhodamine-based fluorescence dyes bearing perfluoroalkyl chains were synthesized. The rhodamines provide a stable fluorescence with high quantum yields, and the attached perfluorinated moieties contribute to its fluorophilicity. The affinity of the F-rhodamines to fluorinated phases was investigated qualitatively in immiscible Langmuir monolayers of stearic and perfluorostearic acid. Afterwards, the ability of the four synthesized F-rhodamine fluorescence dyes to incorporate into lipid membranes was tested with phospholipid/fluorotelomer alcohol mixed GUVs. Finally, the partition coefficients of the F-rhodamines between fluorinated phases and other immiscible organic or aqueous phases, mainly the 1H,1H,2H,2H-perfluoro-1-octanol/water, were determined to quantify their fluorophilicity and distinguish it from lipophilicity for biologically relevant systems.