This paper reports the interaction of nano-clay platelets with a phospholipid in presence of a fluorescence probe. The surface pressure vs. time characteristic studies were done to monitor the progress of reaction. In-situ fluorescence imaging microscope (FIM) was employed to visualize the domain structure formed at the air-water interface with the progress of reaction. At the end of the reaction, clay induced hybrid monolayer at the air-water interface was transferred onto solid substrate to form Langmuir Blodgett (LB) film. Fluorescence spectroscopic studies revealed the close proximity of RhB molecules resulting in the formation of dimeric species.