Analysis of trace low molecular weight (LMW) proteins in serum using the label-free imaging ellipsometry (IE) immunosensor is still a challenge due to the lack of an effective signal amplification strategy and the serious non-specific adsorption. Herein we have developed a sandwich strategy-mediated IE (SSIE) immunosensor to enable the immunodetection of LMW protein biomarkers in serum samples. We have firstly found that the weak binding affinity and the insufficient surface amount density of the ligand are two important factors which hinder the detection of LMW proteins in serum using the IE immunosensor. Then we have deduced that the sandwich strategy can amplify the detection signal of IE and avoid the non-specific adsorption in serum. As a validation of the serological detection of LMW proteins, the SSIE immunosensor has been used to accomplish the quantitative detection of procalcitonin (PCT) in serum. Compared with other PCT analysis methodologies, the SSIE immunosensor enjoys the advantages of simplicity, rapidity, and sufficient sensitivity. Furthermore, we have proposed the criteria to predict the ability of the SSIE immunosensor for the detection of LMW protein biomarkers in serum, which can make the detection of LMW proteins smart and efficient.

After years of development, biosensors based on imaging ellipsometry and biosensors based on total internal reflection imaging ellipsometry have been successfully implemented in various engineering systems. Their experimental setups, detection principles, and biological and clinical applications are briefly reviewed.

A biosensor based on high spatial resolution imaging ellipsometry has been studied to examine its role in the rapid detection and analysis of the tropomyosin allergen existing in crustaceans. This methodology has been established for detection of the tropomyosin allergen and includes ligand screening, the determination of sensitivity, and a comparison with traditional detection methods. Three kinds of monoclonal antibodies (2F9, 4C7, and 2H6) known to have a high bioactivity against the tropomyosin allergen were screened and separately immobilized as ligands on a silicon wafer surface, thus allowing them to capture the tropomyosin allergen. Resulting changes on the wafer surface were visualized in gray scale variation on an ellipsometry image. Images showed that these antibodies are able to recognize the presence of the tropomyosin allergen in shrimp and crab with sensitivity of 1 mg L^–1, and at a detecting time of approximately 30 min for an extracted sample. This preliminary study has shown that the detection of the tropomyosin allergen is rapid and specific when using this type of assay on products containing shrimp and crab. When compared with the enzyme-linked immunosorbent assay, the biosensor based on imaging ellipsometry is able to perform a fast label-free analysis.