A new kind of magnetic/luminescent multifunctional nanoparticles was synthesized by covalently linking multiple carboxyl-functionalized superparamagnetic Fe3O4 nanoparticles and individual amino-functionalized silica-coated fluorescent NaYF4 : Yb,Er up-conversion nanoparticles (UCNPs). The resultant nanocomposites bear active carboxylic and amino groups on the surface that were proved to be chemically active and useful for further facile bioconjugation with biomolecules. The UCNPs in the nanocomposite particles can emit visible light in response to the irradiation by near infrared (NIR) light, enabling the application of the nanocomposites in bioimaging. X-Ray diffraction, infrared spectroscopy, transmission electron microscopy, luminescence spectroscopy, and magnetometry were applied to characterize the multifunctional nanocomposites. The nanocomposites exhibited good superparamagnetic and excellent green up-conversion photoluminescent properties that can be exploited in magnetic separation and guiding as well as bioimaging. Due to the presence of active functional groups on the nanocomposite surface, the Fe3O4/NaYF4 : Yb,Er magnetic/luminescent nanocomposites were successfully conjugated with a protein called transferrin, which specifically recognizes the transferrin receptors overexpressed on HeLa cells, and can be employed for biolabeling and fluorescent imaging of HeLa cells. Because NIR light can penetrate biological samples with good depth without damaging them and can avoid autofluorescence from them, the presence of both NIR-responsive UCNPs and superparamagnetic nanoparticles in the nanocomposite particles will enable the practical application of the nanocomposites in bioimaging and separation.

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Exosomes as nanosized vesicles have been recognized as potential noninvasive biomarkers for early cancer diagnosis. Herein, we presented a sensitive multicolor visual method for exosome detection based on enzyme-induced silver deposition on gold nanorods (Au NRs). To achieve highly sensitive determination of exosomes, hybridization chain reaction (HCR) was employed to introduce more alkaline phosphatase (ALP) for signal amplification. First, exosomes were captured by magnetic bead-labeled CD63 aptamer, and, then, cholesterol-modified DNA probes were spontaneously inserted into the exosomal lipid membrane. The ends of the DNA probes act as the initiator to trigger the HCR for signal amplification. Finally, with the help of HCR, increased sites led to enhanced ALP loading and thus boosted the ascorbic acid generation. Silver ions were reduced by ascorbic acid, and silver shells were formed on Au NRs, giving rise to the blue shift of the longitudinal localized surface plasmon resonance peak. Correspondingly, the concentration of exosomes can be obviously distinguished with naked eyes via the vivid color variation. Due to the dual signal amplification of HCR and metallization of Au NRs, highly sensitive detection for exosomes were realized with detection limits as low as 1.6 × 102 particles/μL by UV–vis spectroscopy and 9 × 103 particles/μL by naked eyes. Compared to the reported colorimetric methods for exosome quantification, visualization based on plentiful color tonalities is the most captivating merit of our approach, and HCR-induced signal amplification highlights the virtue of the strategy. The applicability of the method was validated by the analysis of clinical samples.

An integrated microfluidic concentration gradient chip was developed for generating stepwise concentrations in high-density channels and applied to high-throughput apoptosis analysis of human uterine cervix cancer (HeLa) cells. The concentration gradient was generated by repeated splitting-and-mixing of the source solutions in a radial channel network which consists of multiple concentric circular channels and an increasing number of branch channels. The gradients were formed over hundreds of branches with predictable concentrations in each branch channel. This configuration brings about some distinctive advantages, e.g., more compact and versatile design, high-density of channels and wide concentration ranges. This concentration gradient generator was used in perfusion culture of HeLa cells and a drug-induced apoptosis assay, demonstrated by investigating the single and combined effects of two model anticancer drugs, 5-fluorouracil and Cyclophosphamide, which were divided into 65 concentrations of the two drugs respectively and 65 of their combinatorial concentrations. The gradient generation, the cell culture/stimulation and staining were performed in a single chip. The present device offers a unique platform to characterize various cellular responses in a high-throughput fashion.

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