Calcium (Ca²⁺) ions act as intracellular second messengers in many different signalling processes in plant cells and thus contribute to the amplification step of the signalling pathway and the specificity of the adaptative response. Dynamics of calcium described as spatial and temporal changes of the Ca²⁺ concentrations either in the cytosol and/or in other compartments of the plant cell are now accepted to generate “calcium signatures”, which might be responsible for the initiation of specific downstream events leading to the mounting of an appropriate response. To identify and elucidate the properties of such signatures, highly sensitive and specific methods have been developed and are used to measure and monitor variations in intracellular Ca²⁺ concentrations. Two of these methods, namely bio-luminescence and fluorescence in combination with confocal laser scanning microscopy, are presented.