Understanding the relationships between molecular organization and dynamics of a complex system is very important to understand the photophysical properties of such system. This paper focuses on a novel strategy based on single molecule spectroscopy and single molecule localization microscopy to elucidate the photostability and localization of a fluorophore molecule on a 2D biomembrane. Improvement of in-plane resolution of a signal in a nano-dimension within the diffraction limit has been discussed in a new way. And, how this better in-plane resolution information can be used for precise localization of a single molecule on a 2D system has also been discussed.