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Noble metal nanoparticle-based colorimetric sensors have become powerful tools for the detection of different targets with convenient readout. Among the many types of nanomaterials, noble metal nanoparticles exhibit extraordinary optical responses mainly due to their excellent localized surface plasmon resonance (LSPR) properties. The absorption spectrum of the noble metal nanoparticles was mostly in the visible range. This property enables the visual detection of various analytes with the naked eye. Among numerous color change modes, the way that different concentrations of targets represent vivid color changes has been brought to the forefront because the color distinction capability of normal human eyes is usually better than the intensity change capability. We review the state of the art in noble metal nanoparticle-based multicolor colorimetric strategies adopted for visual quantification by the naked eye. These multicolor strategies based on different means of morphology transformation are classified into two categories, namely, the etching of nanoparticles and the growth of nanoparticles. We highlight recent progress on the different means by which biocatalytic reactions mediated LSPR modulation signal generation and their applications in the construction of multicolor immunoassays. We also discuss the current challenges associated with multicolor colorimetric sensors during actual sample detection and propose the future development of next-generation multicolor qualification strategies.

Herein, we report for the first time a colorful chromogenic substrate, which displays vivid color responses in the presence of different concentration of analytes. Our investigation reveals that the selective shortening of gold nanorods (AuNRs) could generate a series of distinct colors that covers nearly the whole visible range from 400 to 760 nm. These vivid colors can be easily distinguished by the naked eye; as a result, the accuracy of visual inspection could be greatly improved. Next, we demonstrate the utility of AuNRs as multicolor chromogenic substrate to develop a number of colorimetric immunoassay methods, e.g., multicolor enzyme-linked immunosorbent assay (ELISA), multicolor competitive ELISA, and multicolor magnetic immunoassay (MIA). These methods allow us to visually quantify the concentration of a broad range of target molecules with the naked eye, and the obtained results are highly consistent with those state-of-the-art techniques that are tested by the sophisticated apparatus. These multicolor portable and cost-effective immunoassay approaches could be potentially useful for a number of applications, for example, in-home personal healthcare, on-site environmental monitoring, and food inspection in the field.

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The electronic balance, one of the most common pieces of equipment in the laboratory, is normally used to directly measure the weight of a target with high accuracy. However, little attention has been paid to the extension of its applications. In this study, an electronic balance was used as a readout to develop a novel aptasensor for protein quantification for the first time. Thrombin was selected as a model target, and its two aptamers recognizing different sites of the protein were used (one aptamer was immobilized on the surface of magnetic microparticles and the other aptamer was functionalized with platinum nanoparticles). The two aptamers were specifically bound with the thrombin to form a sandwich structure; thus, the platinum nanoparticles were linked to the magnetic microparticles, and they were separated by a magnet easily. The captured platinum nanoparticles effectively catalyzed the decomposition of HO, generating a large volume of O to discharge a certain amount of water in a drainage device, because the pressure in the vial is higher than that outside of the vial. The weight of water was accurately measured by an electronic balance. The weight of water increased with the increasing of the thrombin concentration in the range of 0 to 100 nM with a detection limit of 2.8 nM. This is the first time the use of an electronic balance as a signal readout for biomolecule quantitation in bioassay has been reported.

In this work, 3,3′,5,5′-tetramethylbenzidine(II) (TMB2+), derived from H2O2-horseradish peroxidase (HRP)-3,3′,5,5′-tetramethylbenzidine (H2O2-HRP-TMB) reaction system, was used to etch AuNRs to generate different colors of solution. Many enzyme reactions are involved in the production of H2O2 (e.g., glucose can react with the dissolved oxygen in the presence of glucose oxidase (GOx) to produce H2O2). Given this information, a simple visual biosensor was developed in this study, with glucose as the example target. The detection range of the proposed system varied with the experimental conditions, such as the concentration of GOx and HRP, and enzymatic reaction time. Under the optimized conditions, the longitudinal shift of localized surface plasmon resonances (LSPR) had a linear correlation with the glucose concentration in the range of 0.1~1.0 mM. Meanwhile, the solution displayed a specific color in response to the glucose concentration, thus enabling the visual quantitative detection of glucose at a glance. Compared with the traditional monochromic colorimetry, this multicolor glucose sensor generates various vivid colors, which can be easily distinguished by naked eyes without any sophisticated instrument. Notably, the proposed method has been successfully applied to detect glucose in serum samples with satisfied results.

On the basis of the different diffusivity and existence of electrostatic repulsion between long and short DNA on the negatively changed indium tin oxide (ITO) microelectrode, a simple but sensitive immobilization free solution-phase electrochemical method for DNA methylation detection and inhibitor screening has been developed. Electroactive substance (methylene blue) tagged at the penultimate base T close to the 3'-terminal first, in the absence of DNA methylation, methylene blue-labeled electroactive fragments cannot be generated by Dpn I and results in a weak electrochemical response being detected on the ITO electrode. On the contrary, a remarkable electrochemical response can be achieved by the cleavage in the presence of DNA methylation since methylene blue-labeled electroactive fragments can be generated and aggregate on the ITO electrode. The proposed system does not need complex operation procedures such as bisulfite treatment, PCR amplification, and electrode immobilization. Six ITO microelectrodes had been assembled on the same microchip, which can achieve the parallel detection of the same sample and improve the experimental efficiency of drug screening. The system was used to conveniently and specifically monitor the change of the DNA methylation level with high sensitivity and selectivity. The proposed system has the potential application to screen the drugs as inhibitors on the activity of methyltransferase in the clinic.

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