Fluorescein O-methacrylate was copolymerized with n-butyl acrylate by atom transfer radical polymerization in a "grafting from" reaction with a multifunctional linear macro-initiator to form pH responsive fluorescent bottlebrushes. The brush-like structure of the synthesized macromolecules was confirmed through molecular imaging by atomic force microscopy. NMR spectroscopy showed that the fluorescent units were successfully incorporated into the polymer. The synthesized bottlebrushes displayed highly fluorescent properties under basic conditions, yet showed no fluorescence under neutral or acidic conditions. The fluorescence could be turned on and off by changing the pH of the solution. These bottlebrush molecules could have potential applications in molecular imaging.