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1.
Anal Bioanal Chem ; 416(16): 3775-3783, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38702449

ABSTRACT

Oblique incidence reflectance difference (OIRD) is an emerging technique enabling real-time and label-free detection of bio-affinity binding events on microarrays. The interfacial architecture of the microarray chip is critical to the performance of OIRD detection. In this work, a sensitive label-free OIRD microarray chip was developed by using gold nanoparticle-decorated fluorine-doped tin oxide (AuNPs-FTO) slides as a chip substrate. This AuNPs-FTO chip demonstrates a higher signal-to-noise ratio and improved sensitivity compared to that built on FTO glass, showing a detection limit of as low as 10 ng mL-1 for the model target, HRP-conjugated streptavidin. On-chip ELISA experiments and optical calculations suggest that the enhanced performance is not only due to the higher probe density enabling a high capture efficiency toward the target, but most importantly, the AuNP layer arouses optical interference to improve the intrinsic sensitivity of OIRD. This work provides an effective strategy for constructing OIRD-based microarray chips with enhanced sensitivity, and may help extend their practical applications in various fields.


Subject(s)
Fluorine , Gold , Limit of Detection , Metal Nanoparticles , Tin Compounds , Tin Compounds/chemistry , Gold/chemistry , Metal Nanoparticles/chemistry , Fluorine/chemistry , Microarray Analysis/methods , Enzyme-Linked Immunosorbent Assay/methods
2.
Talanta ; 276: 126201, 2024 Aug 15.
Article in English | MEDLINE | ID: mdl-38718653

ABSTRACT

Oblique-incidence reflectivity difference (OIRD) is a dielectric constant-sensitive technique and exhibits intriguing applications in label-free and high-throughput detection of protein microarrays. With the outstanding advantage of being compatible with arbitrary substrates, however, the effect of the substrate, particularly its dielectric constant on the OIRD sensitivity has not been fully disclosed. In this paper, for the first time we investigated the dependence of OIRD sensitivity on the dielectric constant of the substrate under top-incident OIRD configuration by combining theoretical modeling and experimental evaluation. Optical modeling suggested that the higher dielectric constant substrate exhibits a higher intrinsic sensitivity. Experimentally, three substrates including glass, fluorine-doped tin oxide (FTO) and silicon (Si) with different dielectric constants were selected as microarray substrates and their detection performances were evaluated. In good agreement with the modeling, high dielectric constant Si-based microarray exhibited the highest sensitivity among three chips, reaching a detection limit of as low as 5 ng mL-1 with streptavidin as the model target. Quantification of captured targets on three chips with on-chip enzyme-linked immunosorbent assay (ELISA) further confirmed that the enhanced performance originates from the high dielectric constant enhanced intrinsic OIRD sensitivity. This work thus provides a new way to OIRD-based label-free microarrays with improved sensitivity.


Subject(s)
Protein Array Analysis , Silicon , Tin Compounds , Protein Array Analysis/methods , Silicon/chemistry , Tin Compounds/chemistry , Glass/chemistry , Limit of Detection , Enzyme-Linked Immunosorbent Assay/methods , Fluorine/chemistry , Streptavidin/chemistry
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