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Surface plasmon resonance microscopy (SPRM) has been widely employed in biological fields because of its high spatial resolution and label-free detection modality. In this study, SPRM based on total internal reflection (TIR) is studied via a home-built SPRM system, and the principle of imaging of a single nanoparticle is analyzed as well. By designing a ring filter and combining it with the deconvolution algorithm in Fourier space, the parabolic tail of the nanoparticle image is removed, in which a spatial resolution of 248 nm is obtained. In addition, we also measured the specific binding between the human IgG antigen and goat anti-human IgG antibody using the TIR-based SPRM. The experimental results have proved that the system can image sparse nanoparticles and monitor biomolecular interactions.

Surface plasmon resonance microscopy (SPRM) is a versatile technique for biosensing and imaging that facilitates high-sensitivity, label-free, real-time characterization. To date, SPR technology has been successfully commercialized and its performance has continued to improve. However, this method is inhibited by low spatial resolution and the inability to achieve single-molecule detection. In this report, we present an overview of SPRM research progress in the field of plasma imaging and sensing. A brief review of the technological advances in SPRM is outlined, as well as research progress in important applications. The combination of various new techniques with SPRM is emphasized. Finally, the current challenges and outlook of this technique are discussed.