We report a new method for label-free, sensitive, and facile detection of lead(II) ions (Pb2+) based on an aptamer-target binding event, which is recognized by orientations of liquid crystals (LCs) at aqueous interfaces. The LC film suspended in the aqueous phase demonstrated a homeotropic orientation in contact with a cationic surfactant cetyltrimethylammonium bromide (CTAB) due to self-assembly of CTAB molecules at the aqueous-LC interface. The ordering of LC subsequently changed to planar in the presence of the spinach RNA aptamer (SRNA) due to interactions between CTAB and SRNA. In the presence of the Pb2+ ion, the ordering of LC changed to homeotropic caused by reorganization of CTAB at the LC-aqueous interface. This is due to formation of more stable quadruplex structures of SRNA with Pb2+ ions in comparison to the CTAB-SRNA complex. The sensor exhibited a detection limit of 3 nM, which is well below the permissible limit of Pb2+ in drinking water. Our experiments establish that addition of Pb2+ leads to (i) the formation of Pb2+-SRNA complexes and (ii) a decrease in density of SRNA on the LC interface, but additional studies are required to determine which of these processes underlie the response of the LCs to the Pb2+. We have also demonstrated the potential application of the LC sensor for detection of Pb2+ in tap water. Unlike current laboratory-based heavy-metal-ion assays, this method is comparatively simple in terms of instrumentation, operation, and optical readout.