ABSTRACT
Phase interrogation surface plasmon resonance (P-SPR) biosensors have the highest sensitivity among different types of surface plasmon resonance (SPR) biosensors. However, P-SPR sensors have small dynamic detection range and complex device configuration. To solve these two problems, we designed a multi-channel P-SPR imaging (mcP-SPRi) sensing platform based on a common-path ellipsometry scheme. A wavelength sequential selection (WSS) technique for P-SPRi sensing is developed to select the optimal sensing wavelengths according to different refractive indexes (RIs) of the samples, so the inconsistency of SPR signal response for different biomolecule types caused by the small dynamic detection range is eliminated. And a dynamic detection range of 3.7×10-3 RIU is achieved, which is the largest among the current mcP-SPRi biosensors. Remarkably, the individual SPR phase image acquisition time has been greatly reduced to 1s by using WSS method instead of whole spectrum scanning, which enables the high-throughput mcP-SPRi sensing.
ABSTRACT
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.
Subject(s)
Microscopy , Nanoparticles , Surface Plasmon Resonance/methods , Immunoglobulin GABSTRACT
The widely used surface-based biomolecule sensing scheme has greatly facilitated the investigation of protein-protein interactions in lab-on-a-chip microfluidic systems. However, in most biosensing schemes, the interactions are driven in a passive way: The overall sensing time and sensitivity are totally dependent on the Brownian diffusion process, which has greatly hindered their efficiency, especially at low concentration levels or single-molecule analysis. To break this limitation, we developed an all-optical active method termed optothermophoretic flipping (OTF). It is the first temporal modulated method that biomolecules were enriched and pushed to their counterparts for effective contact via a flipped thermophoresis. As a proof-of-concept experiment, we tested its performance via antibody-antigen binding on a surface plasmon resonance imaging (SPRi) platform. Compared with the interaction solely based on Brownian diffusion, we achieved a 23.6-fold sensitivity increment in biomolecule interactions sensing. This method has opened new opportunities for various biosensing platforms that require high-sensitivity in colloidal sciences and biochemical studies.
