Phase separation and interactions in mixed monolayers of dipalmitoylphosphatidylglycerol (DPPG) with the rhodamine B end-labeled double-hydrophilic block copolymer (DHBC), poly(N,N-dimethylacrylamide)-Nock-poly(N,N-diethylacrylamide) (RhB-PDMA(207)-b-PDEAD(177)). was studied at the air water interface. Surface pressure versus area isotherms indicate that both components behave almost independently. Brewster angle microscopy (BA M) images show a random distribution of liquid condensed (LC) domains of UPPG in an apparent homogeneous matrix of DHBC. excluding the macroscopic phase separation. The laser scanning confocal fluorescence microscopy (LSCFM) of the rhodamine dye at the end of the PUMA chain showed how the DHBC is distributed in Langmuir-Blodgett (LB) mixed monolayers. The high spatial resolution of atomic force microscopy (A FM) combined with the LCSFM images indicates that DHBC incorporates in the expanded phase of DPPG to form mixed domains. being excluded from the condensed regions. Upon compression, nanosized LC domains of DPPG nucleate inside the mixed domains corralled in the nanopatterning, of pure DHBC. The negatively charged polar group of DPPG inhibits rhodamine aggregation, while the long polymer chains promote the formation of corralled nanodomains of DPPG in two dimensions.