Precisely Encoded Barcodes through the Structure-Fluorescence Combinational Strategy: A Flexible, Robust, and Versatile Multiplexed Biodetection Platform with Ultrahigh Encoding Capacities.

With the rapid development of suspension array technology, microbeads-based barcodes as the core element with sufficient encoding capacity are urgently required for high-throughput multiplexed detection. Here, a novel structure-fluorescence combinational encoding strategy is proposed for the first time to establish a barcode library with ultrahigh encoding capacities. Based on the never revealed transformability of the structural parameters (e.g., porosity and matrix component) of mesoporous microbeads into scattering signals in flow cytometry, the enlargement of codes number has been successfully realized in combination with two other fluorescent elements of fluorescein isothiocyanate isomer I (FITC) and quantum dots (QDs). The barcodes are constructed with precise architectures including FITC encapsulated within mesopores and magnetic nanoparticles as well as QDs immobilized on the outer surface to achieve the ultrahigh encoding level of 300 accompanied with superparamagnetism. To the best of knowledge, it is the highest record of single excitation laser-based encoding capacity up to now. Moreover, a ten-plexed tumor markers bioassay based on the tailored-designed barcodes has been evaluated to confirm their feasibility and effectiveness, and the results indicate that the barcodes platform is a promising and robust tool for practical multiplexed biodetection.

Evolution of "On-Barcode" Luminescence Oxygen Channeling Immunoassay by Exploring the Barcode Structure and the Assay System.

The multiplexed luminescence oxygen channeling immunoassay (multi-LOCI) platform we developed recently that combines conventional LOCI and suspension array technology is capable of realizing facile "mix-and-measure" multiplexed assays without tedious washing steps. However, previous work lacks comprehensive studies of the structure-performance relationship of the host-guest-structured barcode, which may obstruct the evolution and further translation of this exciting new technology to practical applications. Accordingly, this work revealed that polyelectrolyte interlayers played a crucial role in tuning the packing density of guest acceptor beads (ABs). More interestingly, we noticed that "sparse" barcodes (barcodes with low ABs packing density) exhibited comparable assay performance with "compact" ones (barcodes with high ABs packing density). The high robustness of barcodes allows for multi-LOCI to be a more universal and flexible assay platform. Furthermore, through optimization of the assay system including the laser power, as well as the concentrations of donor beads and biotinylated detection antibodies, the multi-LOCI platform showed a significant improvement in sensitivity compared with our previous work, with the limit of detection decreasing to as low as ca. 1 pg/mL. Impressively, multi-LOCI that enabled simultaneous detection of multiple analytes exhibited comparable sensitivity with the classical single-plexed LOCI, due to the ingenious structural design of the multi-LOCI barcode and the unique "on-barcode" assay format.

From Low to High Saturation Magnetization in Magnetite Nanoparticles: The Crucial Role of the Molar Ratios Between the Chemicals.

In this study, a comprehensive characterization of iron oxide nanoparticles synthesized by using a simple one-pot thermal decomposition route is presented. In order to obtain monodisperse magnetite nanoparticles with high saturation magnetization, close to the bulk material, the molar ratios between the starting materials (solvents, reducing agents, and surfactants) were varied. Two out of nine conditions investigated in this study resulted in monodisperse iron oxide nanoparticles with high saturation magnetization (90 and 93% of bulk magnetite). The X-ray diffraction analyses along with the inspection of the lattice structure through transmission electron micrographs revealed that the main cause of the reduced magnetization in the other seven samples is likely due to the presence of distortion and microstrain in the particles. Although the thermogravimetric analysis, Raman and Fourier transform infrared spectroscopies confirmed the presence of covalently bonded oleic acid on the surface of all the samples, the particles with higher polydispersity and the lowest surface coating molecules showed the lowest saturation magnetization. Based on the observed results, it could be speculated that the changes in the kinetics of the reactions, induced by varying the molar ratio of the starting chemicals, can lead to the production of the particles with higher polydispersity and/or lattice deformation in their crystal structures. Finally, it was concluded that the experimental conditions for obtaining high-quality iron oxide nanoparticles, particularly the molar ratios and the heating profile, should not be chosen independently; for any specific molar ratio, there may exist a specific heating profile or vice versa. Because this synthetic consideration has rarely been reported in the literature, our results can give insights into the design of iron oxide nanoparticles with high saturation magnetization for different applications.

A facile polymer mediated dye incorporation method for fluorescence encoded microbeads with large encoding capacities.

Here we report a facile dye incorporation method for fluorescence encoded microbeads, which is achieved by tuning the mixed polymer type (blank and dye-labeled polymers) and their doping ratio through electrostatic loading into mesoporous beads. This method is universal to various carriers and could render large encoding capacities.

Ultrabright dye-loaded spherical polyelectrolyte brushes and their fundamental structure-fluorescence tuning principles.

Ultrabright fluorescent particles (UFPs) have attracted increasing attention because of their outstanding signal amplification functions. However, there is still an urgent demand for designing novel UFPs with new components or structures as the existing ones can not satisfy the practical requirements due to their inherent disadvantages. Here we propose a novel ultrabright fluorescent particle platform by doping dyes of 5-aminofluorescein (5-AF) into silica core-based spherical poly (acrylic acid) brushes (SiO2@PAA@5-AF) and discuss their fundamental structure-fluorescence tuning principles. A series of brushes with different polymer chain lengths are successfully synthesized and then loaded with 5-AF through chemical binding. The high loading amount, suitable density or distribution, and enhanced quantum yield (QY) of 5-AF due to the amide bond formation with PAA chains on brushes are concluded as the three major reasons for the ultrabrightness of SiO2@PAA@5-AF. Therefore, a 2350 ± 445 times brighter brush particle in comparison to a single quantum dot (QD) is realized, and a 2.1 ± 0.4 times fluorescence improvement of a brush vs. a QD normalized by volume is also achieved when taking the hydrodynamic diameter into consideration (∼300 nm vs. ∼30 nm). Moreover, the excellent tolerance stabilities in normally applied environments and outstanding label effects to form 4-plexed encoded beads are demonstrated as well. The results in this work strongly indicate a promising potential of SiO2@PAA@5-AF as an ultrabright and stable signal amplification tool for biomedical related sensing, labeling, and biodetection.