Abstract: The invention relates to a new method for generating a luminescent signal, which is based on an enzyme-independent photon emission mechanism (EiPE), whereby luminescent light is generated when a bioluminescent substrate interact with a physical surface in the absence of any catalytic enzyme. In particular embodiments, the method is used to stimulate light emission by fluorescent molecules. The invention relates also to kits for implementing a method according to the present invention and to the use of a bioluminescent substrate, in the presence of a physical surface, to produce a luminescent signal in the absence of enzymatic catalyst.