A lateral flow immunoassay for straightforward determination of fumonisin mycotoxins based on the quenching of the fluorescence of CdSe/ZnS quantum dots by gold and silver nanoparticles.

A lateral flow immunoassay (LFIA) was developed for the determination of fumonisin mycotoxins. The fluorescence of CdSe/ZnS quantum dots (QDs), observed at excitation/emission wavelengths of 365/525 nm, is suppressed by the addition of silver nanoparticles (SNPs) or gold nanoparticles (GNPs) because SNPs overlap the excitation bands of the QDs, and GNPs overlap the emission bands. The fluorescence of the QDs is recovered upon addition of fumonisins, allowing for the sensitive detection in "positive mode" of the target mycotoxin by monitoring the changes of the QDs fluorescence intensity. The SNPs are found to be the most effective quenchers, while the use of GNPs allows for an efficient recovery of fluorescence and can be employed in the LFIA. The method was successfully applied to the fluorometric determination of fumonisins in spiked maize flour samples. The visual detection limit is at the ng·mL-1 level. This is four times lower compared to the colorimetric LFIA based on the use of the conventional gold NPs. Graphical abstract Schematic of the fluorescence quenching lateral flow immunoassay that uses fluorescent quantum dots (QD) and metal nanoparticles (NP) as the quencher: the binding of NP-labelled antibody to the antigen (purple triangle) modulates QD luminescence at the Test line, allowing for 'positive mode' detection of fumonisins. The NP accumulation at Control line assures validity of the test.

Delivery and reveal of localization of upconversion luminescent microparticles and quantum dots in the skin in vivo by fractional laser microablation, multimodal imaging, and optical clearing.

Delivery and spatial localization of upconversion luminescent microparticles [Y2O3:Yb, Er] (mean size ∼1.6 µm) and quantum dots (QDs) (CuInS2/ZnS nanoparticles coated with polyethylene glycol-based amphiphilic polymer, mean size ∼20 nm) inside rat skin was studied in vivo using a multimodal optical imaging approach. The particles were embedded into the skin dermis to the depth from 300 to 500 µm through microchannels performed by fractional laser microablation. Low-frequency ultrasound was applied to enhance penetration of the particles into the skin. Visualization of the particles was revealed using a combination of luminescent spectroscopy, optical coherence tomography, confocal microscopy, and histochemical analysis. Optical clearing was used to enhance the image contrast of the luminescent signal from the particles. It was demonstrated that the penetration depth of particles depends on their size, resulting in a different detection time interval (days) of the luminescent signal from microparticles and QDs inside the rat skin in vivo. We show that luminescent signal from the upconversion microparticles and QDs was detected after the particle delivery into the rat skin in vivo during eighth and fourth days, respectively. We hypothesize that the upconversion microparticles have created a long-time depot localized in the laser-created channels, as the QDs spread over the surrounding tissues.

Synthesis of Hydrophilic CuInS2/ZnS Quantum Dots with Different Polymeric Shells and Study of Their Cytotoxicity and Hemocompatibility.

In this work, there is a detailed description of the whole process of biocompatible CIS/ZnS QDs production. Special attention was paid to the stability of QDs against photooxidation. It was shown that Cu/In ratio greatly affected not only nanocrystals PLQYs but photostability as well. CIS/ZnS QDs with Cu/In = 1:4 ratio showed high photostability under UV illumination both in toluene and aqueous solutions. Meanwhile, photoluminescence of CIS/ZnS QDs with Cu/In = 1:1 ratio was completely quenched after several hours under UV illumination, though their initial QY was as high as 40% with peak maximum at 740 nm. QDs were transferred to water by polymer encapsulation and were subsequently modified with polyethers Jeffamines, cheap analogues of PEG-derivatives. Three types of hydrophilic QDs differing in size, PEG content, and surface charge were obtained for further investigation and comparison of their cytotoxicity and hemocompatibility. It was shown that both leucocytes size distribution and coagulation activation change after introduction of polyethers into QDs polymeric shell, while red blood cells and platelets size distribution as well as hemolysis rate did not show any different results among different QDs and the polymer itself. All three types of QDs showed only slight cytotoxicity. Confocal microscopy proves penetration of hydrophilic CIS/ZnS QDs inside cells, so the low QDs cytotoxocity cannot be explained by low cellular uptake of the QDs and indicated low QDs toxicity in general.

Hydrophilic, bright CuInS2 quantum dots as Cd-free fluorescent labels in quantitative immunoassay.

We report on the synthesis of core-shell CuInS2/ZnS quantum dots (QDs) in organic solution, their encapsulation with a PEG-containing amphiphilic polymer, and the application of the resulting water-soluble QDs as fluorescent label in quantitative immunoassay. By optimizing the methods for core synthesis and shell growth, CuInS2/ZnS QDs were obtained with a quantum yield of 50% on average after hydrophilization. After conjugation with an aflatoxin B1-protein derivative, the obtained QDs were used as fluorescent labels in microplate immunoassay for the quantitative determination of the mycotoxin aflatoxin B1. QDs-based immunoassay showed higher sensitivity compared to enzyme-based immunoassay.

Polymer-coated fluorescent CdSe-based quantum dots for application in immunoassay.

The paper describes all stages of synthesis and characterization of biocompatible CdSe-based core/shell quantum dots (QDs) and their application as fluorescent label for immunoassay. Special attention was focused on development of maleic anhydride-based amphiphilic polymers for QDs solubilization in aqueous media. In this work two PEG-amines were tried for polymer modification: monoamine Jeffamine M 1000 used previously in some researches and diamine Jeffamine ED-2003 applied for the first time for QDs solubilization. The use of different Jeffamines allows us to obtain QDs with carboxyl or amine functional groups available for conjugation. The influence of polymer composition on optical properties of the nanocrystals and their stability in aqueous solutions as well as on their conjugation with biomolecules was studied. QDs with different coatings were used as biolabels in quantitative fluorescence microtiter plate immunoassay and qualitative on-site column test. It was found that quantum dots covered with amphiphilic polymer prepared from poly(maleic anhydride-alt-1-octadecene) and Jeffamine ED-2003 retained up to 90% of their initial brightness, easily conjugated with protein and showed low non-specific adsorption. In optimized conditions the obtained QDs were successfully used for determination of mycotoxin deoxynivalenol in wheat and maize samples by fluorescence microtiter plate immunoassay with an IC50 of 220 µg kg(-1) and by on-site column test with cut-off of 500 µg kg(-1).