The invention discloses a wide-range electron beam excited fluorescence imaging and spectral measurement device and a method thereof. The wide-range electron beam excited fluorescence imaging and spectral measurement device comprises a electron microscope scanning system, a scanning signal generator, a wide-range fluorescence collecting, coupling and transmitting system, a fluorescence intensity detector, a fluorescence spectral detector, a synchronous scanning signal collector and a cooperative control and data processing output system. Since a wide-range fluorescence coupler is introduced, fluorescence signals excited within an imaging view field by the microscope scanning system can be converged and coupled to a fluorescence transmission light path with high collection efficiency, the problems that, to images acquired by wide-range fluorescence scanning imaging, it is difficult to calculate and compare fluorescence excitation intensity or fluorescence yield and fluorescence spectruminformation of different positions according to uniform standards are solved, and wide-range rapid detection and analysis on the basis of electron beam excited fluorescence signals can be completed.