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Sensors 2020, 20, 6890 15 of 23 Sensors 2020, 20, x FOR PEER REVIEW 15 of 23 Figure 6. Schematic illustrations of underling luminescence mechanisms: (a) down-conversion Figure 6. Schematic illustrations of underling luminescence mechanisms: (a) down-conversion nanoparticles are excited in the region where various biomolecules also could be excited and the nanoparticles are excited in the region where various biomolecules also could be excited and the emission of nanoparticles could overlap with autofluorescence; (b) up-conversion nanoparticles are emission of nanoparticles could overlap with autofluorescence; (b) up-conversion nanoparticles are excited in NIR region where biomolecules are not excited, thus there is no autofluorescence of the sample. excited in NIR region where biomolecules are not excited, thus there is no autofluorescence of the sample. Gorris et al. described UCNPs as background-free codes for multiplexed analyses [86]. In the system proposed by the authors, the surface of the UCNPs (NaYF4:Yb,Er and NaYF4:Yb,Tm) is modified Zhang et al. designed upconverting nanocrystals encoded magnetic microspheres (UCNMMs), with a screen layer which consists of different amounts of an organic dye. The absorption spectrum which have the potential application for fast separation and multiplexed immunoassays [90]. Six of this dye overlaps with one of the UCNP bands, thus the emission from this band is re-absorbed unique upconverting nanocrystals with different upconversion emission spectra were synthesised by to different levels by the organic dye. Another emission band is used as a reference. These UCNPs a solvent–thermal process. Both upconverting nanocrystals and magnetic nanoparticles were and organic dye complexes could be used as coding elements which provide ratiometric codes in encapsulated into porous pоly(styrene-co-EGDMA-co-MAA) beads to obtain UCNMMs. Later multiplexed system [88]. UCNMMs were conjugated with goat anti-mouse IgG. For the purpose of detection, FITC-labelled Zhang et al. created upconversion nanobarcodes (UPNBs)—uniform rabbit anti-mouse and IgG PE-labelled goat anti-rabbit IgG antibodies were used. The immunoassay Y O :Yb,Ho,Tm@Y O @SiO (COO-) core/shell nanoparticles, which could be used for optical 23232 results demonstrated no interrelation between upconversion emission, which was used for encoding, encoding in multiplexed immunoassays [89]. The authors demonstrated that by changing the and fluorescence of dyes (FITC, PE), which was used as a reporter. Thus, the authors demonstrated composition of nanoparticles, several different UPNBs can be created because each nanobarcode has an the advantage of upconverting nanomaterials’ utilisation as barcodes in multiplexed immunoassays individual spectrum. Additionally, in order to test synthesised nanobarcodes, UPNBs were conjugated compared with traditional down-conversion materials (fluorescent dyes or QDs). The detection limit with mouse and rabbit antibodies for two-plexed immunoassays. Additionally, microspheres modified of mouse IgG in this model immunoassay was 0.01 ng/mL. Thus, their proposed detection system with goat anti-mouse and goat anti-rabbit antibodies (both cascade blue-labelled) were used for the has huge potential applications in multiplexed immunoassays [90]. immunodetection of potential receptors. In this way, the recognition process was simulated in the In order to improve the diagnostic detection of several viruses such as influenza (A and B), model system. Specific binding between primary antibodies conjugated to UPNBs and secondary respiratory syncytial virus, and adenovirus, Kazakova et al. developed a multiplex serоlogical antibodies conjugated to microspheres were observed, thus providing proof-of-principle of this model microarray immunoassay for the simultaneous detection of serum IgG antibodies against these immunoassay [89]. viruses [91]. Microarray plates were coated with streptavidin and the serum antigens as well as Zhang et al. designed upconverting nanocrystals encoded magnetic microspheres (UCNMMs), negative and positive controls for the samples. Different locations of each of the spotted antigens which have the potential application for fast separation and multiplexed immunoassays [90]. allowed them to implement specificity of this immunoassay. For detection purposes, anti-human Six unique upconverting nanocrystals with different upconversion emission spectra were synthesised IgG-coated NaYF4:Yb,Er upconverting nanoparticles were used. UCNPs were chosen to avoid by a solvent–thermal process. Both upconverting nanocrystals and magnetic nanoparticles autofluоrescence signals from the sample and to increase detection sensitivity. The nanobiosensor were encapsulated into porous poly(styrene-co-EGDMA-co-MAA) beads to obtain UCNMMs. was effectively used for the simultaneous detection of antibodies against seven different viruses. The Later UCNMMs were conjugated with goat anti-mouse IgG. For the purpose of detection, FITC-labelled authors suggest that their multiplexed immunoassay is a promising tool for the diagnostic detection rabbit anti-mouse and IgG PE-labelled goat anti-rabbit IgG antibodies were used. The immunoassay of viral infections in serum samples, and can be utilised in epidemiological and serоprevalence results demonstrated no interrelation between upconversion emission, which was used for encoding, studies [91]. Li et al. demonstrated multiplexed upconversion imaging in vivo by registering distinct lifetimes of UCNPs after excitation at 808 nm [92]. The authors used an NaYF4@NaYbF4@NaYF4:Yb/Tm@NaYF4PDF Image | Multiplexed Nanobiosensors: Current Trends in Early Diagnostics
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