Highly Durable, Stretchable Multielectrode Array for Electro-mechanical Co-stimulation of Cells.

Electro-mechanical co-stimulation of cells can be a useful cue for tissue engineering. However, reliable co-stimulation platforms still have limitations due to low durability of the components and difficulty in optimizing the stimulation parameters. Although various electro-mechanical co-simulation systems have been explored, integrating materials and components with high durability is still limited. To tackle this problem, we designed an electro-mechanical co-stimulation system that facilitates uniaxial cyclic stretching, electrical stimulation, and optical monitoring. This system utilizes a robust and autoclavable stretchable multielectrode array housed within a compact mini-incubator. To illustrate its effectiveness, we conducted experiments that highlighted how electro-mechanical co-stimulation using this system can enhance the maturation of cardiomyocytes derived from human induced pluripotent stem cells. The results showed great potential of our co-stimulation platform as an effective tool for tissue engineering.

Fluorescence-Enhanced Microfluidic Biosensor Platform Based on Magnetic Beads with Highly Stable ZnO Nanorods for Biomarker Detection.

Existing affinity-based fluorescence biosensing systems for monitoring of biomarkers often utilize a fixed solid substrate immobilized with capture probes limiting their use in continuous or intermittent biomarker detection. Furthermore, there have been challenges of integrating fluorescence biosensors with a microfluidic chip and low-cost fluorescence detector. Herein, we demonstrated a highly efficient and movable fluorescence-enhanced affinity-based fluorescence biosensing platform that can overcome the current limitations by combining fluorescence enhancement and digital imaging. Fluorescence-enhanced movable magnetic beads (MBs) decorated with zinc oxide nanorods (MB-ZnO NRs) were used for digital fluorescence-imaging-based aptasensing of biomolecules with improved signal-to-noise ratio. High stability and homogeneous dispersion of photostable MB-ZnO NRs were obtained by grafting bilayered silanes onto the ZnO NRs. The ZnO NRs formed on MB significantly improved the fluorescence signal up to 2.35 times compared to the MB without ZnO NRs. Moreover, the integration of a microfluidic device for flow-based biosensing enabled continuous measurements of biomarkers in an electrolytic environment. The results showed that highly stable fluorescence-enhanced MB-ZnO NRs integrated with a microfluidic platform have significant potential for diagnostics, biological assays, and continuous or intermittent biomonitoring.

Culture-Free Quantification of Bacteria Using Digital Fluorescence Imaging in a Tunable Magnetic Capturing Cartridge for Onsite Food Testing.

Accurate, onsite detection of pathogenic bacteria from food matrices is required to rapidly respond to pathogen outbreaks. However, accurately detecting whole-cell bacteria in large sample volumes without an enrichment step remains a challenge. Therefore, bacterial samples must be concentrated, identified, and quantified. We developed a tunable magnetic capturing cartridge (TMCC) and combined it with a portable digital fluorescence reader for quick, onsite, quantitative detection of Staphylococcus aureus. The TMCC platform integrates an absorption pad impregnated with water-soluble polyvinyl alcohol (PVA) with an injection-molded polycarbonate (PC) plate that has a hard magnet on its back and an acrylonitrile-butadiene-styrene case. An S. aureus-specific antibody conjugated with magnetic nanoparticles was used to concentrate bacteria from a large-volume sample and capture bacteria within the TMCC. The retention time for capturing bacteria on the TMCC was adjusted by controlling the concentration and volume of the PVA solution. Concentrated bacterial samples bound to target-specific aptamer probes conjugated with quantum dots were loaded into the TMCC for a controlled time, followed by attachment of the bacteria to the PC plate and removal of unbound aptamer probes with wash buffer. The captured bacteria were quantified using a digital fluorescence reader equipped with an embedded program that automatically counts fluorescently tagged bacteria. The bacterial count made using the TMCC was comparable to a standard plate count (R2 = 0.9898), with assay sensitivity and specificity of 94.3 and 100%, respectively.

A quorum-based fluorescent probe for imaging pathogenic bacteria.

Imaging of bacterial infections can be used for a wide range of investigations, including diagnosis and pathogenesis of infections, and molecular probes targeting biological processes during infection have been used extensively. However, in vivo visualization of bacteria using molecular probes is still challenging due to the difficulty of directly targeting specific bacteria. Here, we propose a new fluorescent nano-bio probe based on a quorum-sensing (QS) antagonist for imaging drug-resistant bacteria. Because most bacteria use QS-based communication to synchronize the regulation of virulence gene expression during infection, QS-based probes can be used to assess the in vivo localization and distribution of pathogenic bacteria. Our developed fluorescent, quorum-based nano-bio probes (QNBPs) target agr-activation in multiple-drug resistant Staphylococcus aureus (MRSA) and were successfully used for in vivo localization of MRSA in sepsis and dermonecrotic animal infection models. Interestingly, we found that QNBPs can diffuse through biofilms and thus provide a new strategy for detecting MRSA embedded within a biofilm. Our findings suggest that our QNBPs have great potential for directly imaging pathogenic bacteria.

Recent progress, challenges, and prospects of fully integrated mobile and wearable point-of-care testing systems for self-testing.

The rapid growth of research in the areas of chemical and biochemical sensors, lab-on-a-chip, mobile technology, and wearable electronics offers an unprecedented opportunity in the development of mobile and wearable point-of-care testing (POCT) systems for self-testing. Successful implementation of such POCT technologies leads to minimal user intervention during operation to reduce user errors; user-friendly, easy-to-use and simple detection platforms; high diagnostic sensitivity and specificity; immediate clinical assessment; and low manufacturing and consumables costs. In this review, we discuss recent developments in the field of highly integrated mobile and wearable POCT systems. In particular, aspects of sample handling platforms, recognition elements and sensing methods, and new materials for signal transducers and powering devices for integration into mobile or wearable POCT systems will be highlighted. We also summarize current challenges and future prospects for providing personal healthcare with sample-in result-out mobile and wearable POCT.

A fully integrated bacterial pathogen detection system based on count-on-a-cartridge platform for rapid, ultrasensitive, highly accurate and culture-free assay.

Rapid, sensitive and accurate point-of-care-testing (POCT) of bacterial load from a variety of samples can help prevent human infections caused by pathogenic bacteria and mitigate their spreading. However, there is an unmet demand for a POCT device that can detect extremely low concentrations of bacteria in raw samples. Herein, we introduce the 'count-on-a-cartridge' (COC) platform for quantitation of the food-borne pathogenic bacteria Staphylococcus aureus. The system comprised of magnetic concentrator, sensing cartridge and fluorescent image reader with a built-in counting algorithm facilitated fluorescent microscopic bacterial enumeration in user-convenient manner with high sensitivity and accuracy within a couple of hours. The analytical performance of this assay is comparable to that of a standard plate count. The COC assay shows a sensitivity of 92.9% and specificity of 100% performed according to global microbiological criteria for S. aureus which is acceptable below 100 CFU/g in the food matrix. This culture-independent, rapid, ultrasensitive and highly accurate COC assay has great potential for places where prompt bacteria surveillance is in high demand.

A smartphone fluorescence imaging-based mobile biosensing system integrated with a passive fluidic control cartridge for minimal user intervention and high accuracy.

A key challenge for realizing mobile device-based on-the-spot environmental biodetection is that a biosensor integrated with a fluid handling sensor cartridge must have acceptable accuracy comparable to that of conventional standard analytical methods. Furthermore, the user interface must be easy to operate, technologically plausible, and concise. Herein, we introduced an advanced smartphone imaging-based fluorescence microscope designed for Hg2+ monitoring by utilizing a biosensor cartridge that reduced user intervention via time-sequenced passive fluid handling. The cartridge also employed a metal-nanostructured plastic substrate for complementing the fluorescence signal output; this helped the realization of high-accuracy detection, in which a ratiometric dual-wavelength detection method was applied. Using 30 samples of Hg2+-spiked wastewater, we showed that our device, which has a detection limit of ∼1 pM, can perform analytical assays accurately. The detection results from our method were in good linearity and agreement with those of conventional standard methods. We conclude that the integration of a simple-to-use biosensor cartridge, fluorescence signal-enhancing substrate, dual-wavelength detection, and quantitative image data processing on a smartphone has great potential to make any population accessible to small-molecule detection, which has been performed in centralized laboratories for environmental monitoring.

Culture-free, highly sensitive, quantitative detection of bacteria from minimally processed samples using fluorescence imaging by smartphone.

A critical unmet need in the diagnosis of bacterial infections, which remain a major cause of human morbidity and mortality, is the detection of scarce bacterial pathogens in a variety of samples in a rapid and quantitative manner. Herein, we demonstrate smartphone-based detection of Staphylococcus aureus in a culture-free, rapid, quantitative manner from minimally processed liquid samples using aptamer-functionalized fluorescent magnetic nanoparticles. The tagged S. aureus cells were magnetically captured in a detection cassette, and then fluorescence was imaged using a smartphone camera with a light-emitting diode as the excitation source. Our results showed quantitative detection capability with a minimum detectable concentration as low as 10 cfu/ml by counting individual bacteria cells, efficiently capturing S. aureus cells directly from a peanut milk sample within 10 min. When the selectivity of detection was investigated using samples spiked with other pathogenic bacteria, no significant non-specific detection occurred. Furthermore, strains of S. aureus from various origins showed comparable results, ensuring that the approach can be widely adopted. Therefore, the quantitative fluorescence imaging platform on a smartphone could allow on-site detection of bacteria, providing great potential assistance during major infectious disease outbreaks in remote and resource-limited settings.

The Pine-Needle-Inspired Structure of Zinc Oxide Nanorods Grown on Electrospun Nanofibers for High-Performance Flexible Supercapacitors.

Flexible supercapacitors with high electrochemical performance and stability along with mechanical robustness have gained immense attraction due to the substantial advancements and rampant requirements of storage devices. To meet the exponentially growing demand of microsized energy storage device, a cost-effective and durable supercapacitor is mandatory to realize their practical applications. Here, in this work, the fabrication route of novel electrode materials with high flexibility and charge-storage capability is reported using the hybrid structure of 1D zinc oxide (ZnO) nanorods and conductive polyvinylidene fluoride-tetrafluoroethylene (P(VDF-TrFE)) electrospun nanofibers. The ZnO nanorods are conformably grown on conductive P(VDF-TrFE) nanofibers to fabricate the light-weighted porous electrodes for supercapacitors. The conductive nanofibers acts as a high surface area scaffold with significant electrochemical performance, while the addition of ZnO nanorods further enhances the specific capacitance by 59%. The symmetric cell with the fabricated electrodes presents high areal capacitance of 1.22 mF cm-2 at a current density of 0.1 mA cm-2 with a power density of more than 1600 W kg-1 . Furthermore, these electrodes show outstanding flexibility and high stability with 96% and 78% retention in specific capacitance after 1000 and 5000 cycles, respectively. The notable mechanical durability and robustness of the cell acquire both good flexibility and high performance.

A smartphone imaging-based label-free and dual-wavelength fluorescent biosensor with high sensitivity and accuracy.

The accuracy of a bioassay based on smartphone-integrated fluorescent biosensors has been limited due to the occurrence of false signals from non-specific reactions as well as a high background and low signal-to-noise ratios for complementary metal oxide semiconductor image sensors. To overcome this problem, we demonstrate dual-wavelength fluorescent detection of biomolecules with high accuracy. Fluorescent intensity can be quantified using dual wavelengths simultaneously, where one decreases and the other increases, as the target analytes bind to the split capture and detection aptamer probes. To do this, we performed smartphone imaging-based fluorescence microscopy using a microarray platform on a substrate with metal-enhanced fluorescence (MEF) using Ag film and Al2O3 nano-spacer. The results showed that the sensitivity and specificity of the dual-wavelength fluorescent quantitative assay for the target biomolecule 17-ß-estradiol in water were significantly increased through the elimination of false signals. The detection limit was 1pg/mL and the area under the receiver operating characteristic curve of the proposed assay (0.922) was comparable to that of an enzyme-linked immunosorbent assay (0.956) from statistical accuracy tests using spiked wastewater samples. This novel method has great potential as an accurate point-of-care testing technology based on mobile platforms for clinical diagnostics and environmental monitoring.

Seesawed fluorescence nano-aptasensor based on highly vertical ZnO nanorods and three-dimensional quantitative fluorescence imaging for enhanced detection accuracy of ATP.

Probe-mediated fluorescence biosensing methods based on spectrophotometry still have limitations such as detection inaccuracy caused by the occurrence of false signals and lack of simultaneous qualitative and quantitative read-outs with an ultra-low detection limit. Herein, we describe a novel seesawed fluorescence detection strategy based on dual-colour imaging-based quantitation in which the green fluorescence of the capture aptamer decreases and the red fluorescence of the detection aptamer increases simultaneously upon their respective interactions with the target biomolecule. This approach enhances detection accuracy through facilitating identification of probable false-positives in biological samples. Furthermore, combining the seesawed detection scheme with three-dimensional imaging of fluorescence signal enhanced by highly vertical ZnO nanorods increases signal-to-noise ratio, which addresses the limited performance of digital cameras and, in turn, enhances sensitivity and dynamic range. This simple, robust, scalable, imaging-based and label-free fluorescence method allows highly specific and sensitive quantification of biomolecules with excellent reliability.

Real-time label-free quantitative fluorescence microscopy-based detection of ATP using a tunable fluorescent nano-aptasensor platform.

Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (rGO) surfaces, thereby providing cytotoxicity assessment. Compared with conventional fluorescence spectrometry methods, our highly efficient, universally applicable, and rational approach will facilitate broader implementation of imaging-based biosensing platforms for the quantitative evaluation of a range of target molecules.

Pharmacophore based approach to design inhibitors in crustaceans: an insight into the molt inhibition response to the receptor guanylyl cyclase.

The first set of competitive inhibitors of molt inhibiting hormone (MIH) has been developed using the effective approaches such as Hip-Hop, virtual screening and manual alterations. Moreover, the conserved residues at 71 and 72 positions in the molt inhibiting hormone is known to be significant for selective inhibition of ecdysteroidogenesis; thus, the information from mutation and solution structure were used to generate common pharmacophore features. The geometry of the final six-feature pharmacophore was also found to be consistent with the homology-modeled MIH structures from various other decapod crustaceans. The Hypo-1, comprising six features hypothesis was carefully selected as a best pharmacophore model for virtual screening created on the basis of rank score and cluster processes. The hypothesis was validated and the database was virtually screened using this 3D query and the compounds were then manually altered to enhance the fit value. The hits obtained were further filtered for drug-likeness, which is expressed as physicochemical properties that contribute to favorable ADME/Tox profiles to eliminate the molecules exhibit toxicity and poor pharmacokinetics. In conclusion, the higher fit values of CI-1 (4.6), CI-4 (4.9) and CI-7 (4.2) in conjunction with better pharmacokinetic profile made these molecules practically helpful tool to increase production by accelerating molt in crustaceans. The use of feeding sub-therapeutic dosages of these growth enhancers can be very effectively implemented and certainly turn out to be a vital part of emerging nutritional strategies for economically important crustacean livestock.