Surface plasmon resonance (SPR) sensing technology is widely used in the field of biosensors due to its non-marking, high-sensitivity, and non-invasive characteristics. However, SPR technology is still limited to sensing analysis in twodimensional plane, axial detection, as the key of SPR application in three-dimensional medium spatial detection, has not been well studied and solved. In an angle-interrogation SPR sensing system, the spatial characteristics of evanescent wave-dielectric interaction at multiple wavelengths are studied, and the factors affecting the spatial distribution of surface plasmon resonance are also analyzed. An axial spatial resolution method based on the particle swarm optimization (PSO) algorithm with multi-wavelength angle-interrogation structure is proposed, the refractive index distribution in axial space is determined by analyzing the characteristic SPR signal. In addition, the calculation and analysis of the applicable range of wavelengths are carried out. In the reliable spectral range of the incident light wavelength of 600-900 nm, the average error of the axial refractive index spatial resolution increases from 10<sup>-5</sup> RIU to 10<sup>-4</sup> RIU as the number of axial layers increases. The proposed multi-wavelength angle modulation structure analysis method based on PSO algorithm extends the SPR detection range from two-dimensional plane to three-dimensional space, which provides a new and promising analysis model for molecular biology.