Abstract: A sensor based on a molecularly-imprinted sol-gel material was developed to detect fluorene in aqueous solutions and the gas phase. The design was based on a hybrid non-covalent and covalent molecular imprinting scheme that resulted in the ability to chemically attach an environmentally sensitive NBD fluorophore adjacent an imprinted binding pocket. Analyte binding was detected by a change in NBD fluorescence induced by a change in polarity. The imprinted sol-gels were tested using direct binding assays to evaluate the strength and specificity of the binding interactions. Molecularly imprinted sol-gel sensing materials were deposited as films on glass supports and showed a 37.1 ± 0.7% increase in NBD fluorescence when exposed to 100 ppb aqueous solutions of fluorene. Calibration curves showed a logarithmic response to fluorene with an average half-saturation point of 0.80 ppb. Other factors including the kinetics of the response, non-specific binding, and sensor reversibility are discussed.