A Localized surface plasmon resonance (LSPR) sensor integrated automated microfluidic system for multiplex inflammatory biomarker detection.

Inflammation is a complex biological response of the human body to external or internal stimuli, such as invading pathogens, defective cells, or irritating substances. One important indicator of inflammatory conditions or the progress of various diseases, such as cancer, cardiovascular diseases, neurological diseases, connective tissue diseases, sepsis, or Alzheimer's disease, is the concentration level of inflammatory biomarkers, including immunoglobulins, cytokines, and C-reactive protein (CRP). Since inflammatory biomarkers are highly correlated with each other, it is important to measure them simultaneously. To enable continuous and dynamic inflammatory biomarker detection, we utilized localized surface plasmon resonance (LSPR) to perform label-free molecule sensing. Since the LSPR sensing mechanism requires only a small sensing area with simplified optical setup, it can be easily integrated with a microfluidic device. To simplify reagent operation complexity, we developed an automated microfluidic control system to control reagent guiding and switching in the immunoassay with less manual processes and potential operation errors. Our results successfully demonstrated multiplex IgG, TNF-α, and CRP measurement with only 60 µL assay volume and 3.5 h assay time. In each test, 20 sensing spot measurements under four different reagent conditions can be performed. Overall, we envision that the LSPR sensor integrated automated microfluidic control system could perform rapid, multiplex, and multiparallel continuous inflammatory biomarker detection, which would be beneficial for various applications, such as immune status monitoring, diagnosis and prognosis of inflammatory diseases.

A Large-Area Nanoplasmonic Sensor Fabricated by Rapid Thermal Annealing Treatment for Label-Free and Multi-Point Immunoglobulin Sensing.

Immunoglobulins are important biomarkers to evaluate the immune status or development of infectious diseases. To provide timely clinical treatments, it is important to continuously monitor the level of multiple immunoglobulins. Localized surface plasmon resonance (LSPR)-based nanoplasmonic sensors have been demonstrated for multiplex immunoglobulins detection. However, the sensor fabrication process is usually slow and complicated, so it is not accessible for large-area and batch fabrication. Herein, we report a large-area (2 cm × 2 cm) nanofabrication method using physical vapor deposition followed by a rapid thermal annealing treatment. To optimize the sensor performance, we systematically characterized three fabrication conditions, including (1) the deposition thickness; (2) the maximum annealing temperature, and (3) the annealing time. The corresponding absorbance spectrum profile and surface morphology of the nanostructures were observed by a UV-VIS spectrometer and atomic force microscopy. We then tested the sensitivity of the sensor using a glucose solution at different concentrations. The results showed that the sensor with 10 nm gold deposition thickness under 5-min 900 °C rapid thermal annealing can achieve the highest sensitivity (189 nm RIU-1). Finally, we integrated this nanoplasmonic sensor with a microchannel and a motorized stage to perform a 10-spot immunoglobulin detection in 50 min. Based on its real-time, dynamic and multi-point analyte detection capability, the nanoplasmonic sensor has the potential to be applied in high-throughput or multiplex immunoassay analysis, which would be beneficial for disease diagnosis or biomedical research in a simple and cost-effective platform.