In-Channel Printing-Device Opening Assay for Micropatterning Multiple Cells and Gene Analysis

Herein we report an easy but versatile method for patterning different cells on a single substrate by using a microfluidic approach that allows not only spatial and temporal control of multiple microenvironments but also retrieval of specific treated cells to profile their expressed genetic information at around 10-cell resolution. By taking advantages of increased surface area of gold nanoparticles on a poly­(dimethylsiloxane) (PDMS) coated substrate, cell adhesive-promotive protein, human fibronectin (hFN) can be significantly accumulated on designed regions where cells can recognize the protein and spread out. Moreover, the whole device can be easily opened by hand without any loss of patterned cells which could be retrieved by mouth-pipet. Consequently, we demonstrate the possibility of analyzing the difference of gene expression patterns between wild type MCF-7 cell and MCF/Adr (drug-resistant cell line) from less than 400 cells in total for a single comprehensive assay, including parallel experiments, controls, and multiple dose treatments. Certain genes, especially the P-glycoprotein coding gene (ABCB1), show high expression level in resistant cells compared with the wild type, suggesting a possible pathway that may contribute to the antidrug mechanism

Chemical Etching of Bovine Serum Albumin-Protected Au25 Nanoclusters for Label-Free and Separation-Free Ratiometric Fluorescent Detection of Tris(2-carboxyethyl)phosphine

This study describes a novel ratiometric fluorescent sensor based on chemical etching of gold nanocluster (GNCs) for label-free, separation-free determination of tris­(2-carboxyethyl)­phosphine (TCEP). TCEP was discovered to exhibit unusual chemical behavior toward fluorescent gold nanoclusters: it quenched the red fluorescent emission of the bovine serum album (BSA)-protected GNCs (GNCs@BSA) and simultaneously restored the blue fluorescent emission of the dityrosine (diTyr) residues of the BSA ligand. The TCEP-induced quenching of the fluorescent GNCs@BSA was investigated with the UV–vis adsorption spectrum, the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), inductively coupled plasma-mass spectrometry (ICP-MS) and X-ray photoelectron spectroscopy (XPS), revealing the chemical etching of the gold(0) core of the GNCs@BSA by TCEP. Furthermore, the ratio of the blue fluorescence intensity of the diTyr to the red fluorescence intensity of the GNCs@BSA was found to be dependent on TCEP concentration and showed a linear relationship in the TCEP concentration range of 500 nM to 50, 000 nM (<i>R</i>² = 0.9943) with a limit of detection (LOD) of 130 nM, achieving the higher sensitivity over previous reports. This ratiometric sensor also showed superior selectivity for TCEP over certain common interferences including glutathione, 20 kinds of natural amino acids, and the oxidized form of TCEP. With the developed ratiometric method, the deproteinized human serum samples spiked with TCEP were analyzed with satisfactory results. In addition, it is worth noting that compared with conventional ratiometric fluorescent sensors, the ratiometric sensor developed in this study does not require external fluorophores, avoiding the additional derivation procedures

Intracellular MicroRNA Imaging and Specific Discrimination of Prostate Cancer Circulating Tumor Cells Using Multifunctional Gold Nanoprobe-Based Thermophoretic Assay

Circulating tumor cells (CTCs) have emerged as powerful biomarkers for diagnosis of prostate cancer. However, the effective identification and concurrently accurate imaging of CTCs for early screening of prostate cancer have been rarely explored. Herein, we reported a multifunctional gold nanoprobe-based thermophoretic assay for simultaneous specific distinguishing of prostate cancer CTCs and sensitive imaging of intracellular microRNA (miR-21), achieving the rapid and precise detection of prostate cancer. The multifunctional gold nanoprobe (GNP-DNA/Ab) was modified by two types of prostate-specific antibodies, anti-PSMA and anti-EpCAM, which could effectively recognize the targeting CTCs, and meanwhile linked double-stranded DNA for further visually imaging intracellular miR-21. Upon the specific internalization of GNP-DNA/Ab by PC-3 cells, target aberrant miR-21 could displace the signal strand to recover the fluorescence signal for sensitive detection at the single-cell level, achieving single PC-3 cell imaging benefiting from the thermophoresis-mediated signal amplification procedure. Taking advantage of the sensitive miR-21 imaging performance, GNP-DNA/Ab could be employed to discriminate the PC-3 and Jurkat cells because of the different expression levels of miR-21. Notably, PC-3 cells were efficiently recognized from white blood cells, exhibiting promising potential for the early diagnosis of prostate cancer. Furthermore, GNP-DNA/Ab possessed good biocompatibility and stability. Therefore, this work provides a great tool for aberrant miRNA-related detection and specific discrimination of CTCs, achieving the early and accurate diagnosis of prostate cancer

A cloud-based X73 ubiquitous mobile healthcare system: design and implementation

Based on the user-centric paradigm for next generation networks, this paper describes a ubiquitous mobile healthcare (uHealth) system based on the ISO/IEEE 11073 personal health data (PHD) standards (X73) and cloud computing techniques. A number of design issues associated with the system implementation are outlined. The system includes a middleware on the user side, providing a plug-and-play environment for heterogeneous wireless sensors and mobile terminals utilizing different communication protocols and a distributed "big data" processing subsystem in the cloud. The design and implementation of this system are envisaged as an efficient solution for the next generation of uHealth systems

Hidden Dityrosine Residues in Protein-Protected Gold Nanoclusters

The protein ligand shells of fluorescent protein-protected gold nanoclusters play an important role in the physiochemical properties and sensing applications of the nanoclusters. Recently, more and more attention has been paid to the investigation of the changes in the protein structure elements induced by the introduction of the nanoclusters in the proteins. In this work, the strategy of removal of the encapsulated gold nanoclusters from the protein ligand cages has been proposed, producing the “hollow” (or possibly “imprinted”) proteins for investigations for the first time. Nontoxic cysteamine was used as the etchant of the gold nanoclusters. With bovine serum albumin, lysozyme, and ovalbumin as model proteins, it was found that the luminescent dityrosine cross-links exist in the protein-protected gold nanoclusters, however, inner filter effect caused by the gold nanoclusters hide them

Electrochemically Mediated Surface-Initiated de Novo Growth of Polymers for Amplified Electrochemical Detection of DNA

The development of convenient and efficient strategies without involving any complex nanomaterials or enzymes for signal amplification is of great importance in bioanalytical applications. In this work, we report the use of electrochemically mediated surface-initiated atom transfer radical polymerization (SI-eATRP) as a novel amplification strategy based on the de novo growth of polymers (dnGOPs) for the electrochemical detection of DNA. Specifically, the capture of target DNA (tDNA) by the immobilized peptide nucleic acid (PNA) probes provides a high density of phosphate groups for the subsequent attachment of ATRP initiators onto the electrode surface by means of the phosphate-Zr⁴⁺-carboxylate chemistry, followed by the de novo growth of electroactive polymer via the SI-eATRP. De novo growth of long polymeric chains enables the labeling of numerous electroactive probes, which in turn greatly improves the electrochemical response. Moreover, it circumvents the slow kinetics and poor coupling efficiency encountered when nanomaterials or preformed polymers are used and features sufficient flexibility and simplicity in controlling the degree of signal amplification. Under optimal conditions, it allows a highly sensitive and selective detection of tDNA within a broad linear range from 0.1 fM to 0.1 nM (<i>R</i>² = 0.996), with the detection limit down to 0.072 fM. Compared with the unamplified method, more than 1.2 × 10⁶-fold sensitivity improvement in DNA detection can be achieved. By virtue of its simplicity, high efficiency, and cost-effectiveness, the proposed dnGOPs-based signal amplification strategy holds great potential in bioanalytical applications for the sensitive detection of biological molecules

Ion Permeability of Polydopamine Films Revealed Using a Prussian Blue-Based Electrochemical Method

Polydopamine (PDA) is fast becoming a popular surface modification technique. Detailed understanding of the ion permeability properties of PDA films will improve their applications. Herein, we report for the first time the thickness-independent ion permeability of PDA films using a Prussian blue (PB)-based electrochemical method. In this method, PDA films are deposited via ammonium persulfate-induced dopamine polymerization onto a PB electrode. The ion permeability of the PDA films can thus be detected by observing the changes in electrochemical behaviors of the PB coated by PDA films. On the basis of this method, it was unexpectedly found that the PDA films with thickness greater than 45 nm (e.g., ∼60 and ∼113 nm) can exhibit pH-switchable but thickness-insensitive permeability to monovalent cations such as potassium and sodium ions. These observations clearly indicate the presence of a continuous network of interconnected intermolecular voids within PDA films, regardless of film thickness

Fully Integrated Ratiometric Fluorescence Enrichment Platform for High-Sensitivity POC Testing of Salivary Cancer Biomarkers

The point-of-care (POC) testing of cancer biomarkers in saliva with both high sensitivity and accuracy remains a serious challenge in modern clinical medicine. Herein, we develop a new fully integrated ratiometric fluorescence enrichment platform that utilizes acoustic radiation forces to enrich dual-emission sandwich immune complexes for a POC visual assay. As a result, the color signals from red and green fluorescence (capture probe and report probe, respectively) are enhanced by nearly 10 times, and colorimetric sensitivity is effectively improved. When illuminated using a portable UV lamp, the fluorescence color changing from red to green can be clearly seen with the naked eye, which allows a semiqualitative assessment of the carcinoembryonic antigen (CEA) level. In combination with a homemade smartphone-based portable device, cancer biomarkers like CEA are quantified, achieving a limit of detection as low as 0.012 ng/mL. We also directly quantify CEA in human saliva samples to investigate the reliability of this fully integrated platform, thus validating the usefulness of the proposed strategy for clinical diagnosis and home monitoring of physical conditions

Synergistic Inhibitory Effect of GQDs–Tramiprosate Covalent Binding on Amyloid Aggregation

Inhibiting the amyloid aggregation is considered to be an effective strategy to explore possible treatment of amyloid-related diseases including Alzheimer’s disease, Parkinson’s disease, and type II diabetes. Herein, a new high-efficiency and low-cytotoxicity Aβ aggregation inhibitors, GQD-T, was designed through the combination of two Aβ aggregation inhibitors, graphene quantum dots (GQDs) and tramiprosate. GQD-T showed the capability of efficiently inhibiting the aggregation of Aβ peptides and rescuing Aβ-induced cytotoxicity due to the synergistic effect of the GQDs and tramiprosate. In addition, the GQD-T has the characteristics of low toxicity and great biocompatibility. It is believed that GQD-T may be a potential candidate for an Alzheimer’s drug and this work provides a new strategy for exploring Aβ peptide aggregation inhibitors

Fully Integrated Ratiometric Fluorescence Enrichment Platform for High-Sensitivity POC Testing of Salivary Cancer Biomarkers

The point-of-care (POC) testing of cancer biomarkers in saliva with both high sensitivity and accuracy remains a serious challenge in modern clinical medicine. Herein, we develop a new fully integrated ratiometric fluorescence enrichment platform that utilizes acoustic radiation forces to enrich dual-emission sandwich immune complexes for a POC visual assay. As a result, the color signals from red and green fluorescence (capture probe and report probe, respectively) are enhanced by nearly 10 times, and colorimetric sensitivity is effectively improved. When illuminated using a portable UV lamp, the fluorescence color changing from red to green can be clearly seen with the naked eye, which allows a semiqualitative assessment of the carcinoembryonic antigen (CEA) level. In combination with a homemade smartphone-based portable device, cancer biomarkers like CEA are quantified, achieving a limit of detection as low as 0.012 ng/mL. We also directly quantify CEA in human saliva samples to investigate the reliability of this fully integrated platform, thus validating the usefulness of the proposed strategy for clinical diagnosis and home monitoring of physical conditions