Lipid reorganization induced morphology alteration in asymmetric bilayers prepared by Langmuir-Blodgett (LB)/ Langmuir-Schaeffer (LS) method was directly visualized by means of the atomic force microscopy (AFM) and fluorescence imaging. The LB monolayer in the bilayer system was prepared using binary mixture of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dipalmitoyl -sn-glycero-3-phosphocholine (DPPC) in 1:1 molar ratio, with 0.2 mol% Texas Red 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine triethylammonium salt (TR-DHPE dye). The shapes of the flower like DPPC-enriched domains were found to be dependent on the preparing temperatures. These domains were well maintained after the LS layer of DOPC was deposited. The contrast of these domains has changed from the original dark to bright, and turned dark again after the sample stayed in water for several hours. Along with the contrast reverse, a third phase was observed to separate out within the domains, exhibiting a much taller height than the outside-domain areas. Mechanical properties of these regions were detected by means of force spectroscopy. The increased heights within DPPC domains and the rapid contrast reverse suggest that not only DOPC but also DPPC lipids have reorganized in the LB/LS bilayer, stimulated the morphology changes.