A ratiometric fluorescence sensor for Be 2+ has been fabricated via alternate assembly of 2-(3,6-disulfo-8-hydroxynaphthylazo)-1,8-dihydroxynaphthalene-3,6-disulfonate (Beryllon II) and MgAl-LDH nanosheets on quartz substrates using the layer-by-layer (LBL) deposition technique. UV–vis absorption and the fluorescence emission spectroscopy indicate a stepwise and regular growth of the Beryllon II/LDH UTFs upon increasing deposition cycle. The film of Beryllon II/LDH possesses a periodic layered structure perpendicular to the substrate revealed by X-ray diffraction and scanning electron microscopy. Atomic force microscopy images show that the film surface is continuous and uniform. The Beryllon II/LDH UTFs display ratiometric fluorescence response for Be 2+ with a linear response range in 1.0×10 −7 –1.9×10 −6 molL −1 and a detection limit of 4.2×10 −9 molL −1 . Furthermore, the ratiometric sensor exhibits good repeatability, high stability (thermal, storage and mechanical) as well as excellent selectivity toward Be 2+ . XPS and Raman measurements demonstrate that the specific response of the sensor is attributed to the coordination between Be 2+ and Beryllon II in the UTF. The Beryllon II/LDH UTFs in this work can be potentially used as a chemosensor for the detection of Be 2+ in the environmental and biomedical field.