ABSTRACT
Here, we apply rigorous coupled-wave theory to analyze the optical phase imaging performance of scanning confocal surface plasmon microscope. The scanning confocal surface plasmon resonance microscope is an embedded interferometric microscope interfering between two integrated optical beams. One beam is provided by the central part around the normal incident angle of the back focal plane, and the other beam is the incident angles beyond the critical angle, exciting the surface plasmon. Furthermore, the two beams can form an interference signal inside a confocal pinhole in the image plane, which provides a well-defined path for the surface plasmon propagation. The scanning confocal surface plasmon resonance microscope operates by scanning the sample along the optical axis z, so-called V(z). The study investigates two imaging modes: non-quantitative imaging and quantitative imaging modes. We also propose a theoretical framework to analyze the scanning confocal surface plasmon resonance microscope compared to non-interferometric surface plasmon microscopes and quantify quantitative performance parameters including spatial resolution and optical contrast for non-quantitative imaging; sensitivity and crosstalk for quantitative imaging. The scanning confocal SPR microscope can provide a higher spatial resolution, better sensitivity, and lower crosstalk measurement. The confocal SPR microscope configuration is a strong candidate for high throughput measurements since it requires a smaller sensing channel than the other SPR microscopes.
ABSTRACT
Surface plasmon microscopy has been of interest to the science and engineering community and has been utilized in broad aspects of applications and studies, including biochemical sensing and biomolecular binding kinetics. The benefits of surface plasmon microscopy include label-free detection, high sensitivity, and quantitative measurements. Here, a theoretical framework to analyze and compare several non-interferometric surface plasmon microscopes is proposed. The scope of the study is to (1) identify the strengths and weaknesses in each surface plasmon microscopes reported in the literature; (2) quantify their performance in terms of spatial imaging resolution, imaging contrast, sensitivity, and measurement accuracy for quantitative and non-quantitative imaging modes of the microscopes. Six types of non-interferometric microscopes were included in this study: annulus aperture scanning, half annulus aperture scanning, single-point scanning, double-point scanning, single-point scanning, at 45 degrees azimuthal angle, and double-point scanning at 45 degrees azimuthal angle. For non-quantitative imaging, there is a substantial tradeoff between the image contrast and the spatial resolution. For the quantitative imaging, the half annulus aperture provided the highest sensitivity of 127.058 rad/µm2 RIU-1, followed by the full annulus aperture of 126.318 rad/µm2 RIU-1. There is a clear tradeoff between spatial resolution and sensitivity. The annulus aperture and half annulus aperture had an optimal resolution, sensitivity, and crosstalk compared to the other non-interferometric surface plasmon resonance microscopes. The resolution depends strongly on the propagation length of the surface plasmons rather than the numerical aperture of the objective lens. For imaging and sensing purposes, the recommended microfluidic channel size and protein stamping size for surface plasmon resonance experiments is at least 25 µm for accurate plasmonic measurements.
