Novel Digital SERS-Microfluidic Chip for Rapid and Accurate Quantification of Microorganisms.

In this work, we present a simple and novel digital surface-enhanced Raman spectroscopy (SERS)-microfluidic chip designed for the rapid and accurate quantitative detection of microorganisms. The chip employs a high-density inverted pyramid microcavity (IPM) array to separate and isolate microbial samples. The presence or absence of target microorganisms is determined by scanning the IPM array using SERS and identifying the characteristic Raman bands. This approach allows for the "digitization" of the SERS response of each IPM, enabling quantification through the application of mathematical statistical techniques. Significantly, precise quantitative detection of yeast was achieved within a concentration range of 106-109 cells/mL, with the maximum relative standard deviation from the concentration calibrated by the cultivation method being 5.6%. This innovative approach efficiently addresses the issue of irregularities in SERS quantitative detection, which arises due to fluctuations in SERS intensity and poor reproducibility. We strongly believe that this digital SERS-microfluidic chip holds immense potential for diverse applications in the rapid detection of various microorganisms, including pathogenic bacteria and viruses.

Clinical characteristics and therapeutic strategy of granulomatous mastitis accompanied by Corynebacterium kroppenstedtii: a retrospective cohort study.

BACKGROUND: Increasing evidence has suggested that Corynebacterium kroppenstedtii is associated with some cases of granulomatous mastitis, mostly based on pathology or microbiology. We aimed to identify the clinical characteristics and treatment regimens for granulomatous mastitis with Corynebacterium kroppenstedtii infection. Understanding these clinical features is essential for patient care. METHODS: We retrospectively collected data on 201 patients who were pathologically diagnosed with granulomatous mastitis and had microbiological results of either Corynebacterium kroppenstedtii or no bacterial growth and recorded and analysed their demographics, clinical characteristics, and clinical outcomes. RESULTS: There were 107 patients in the CK group and 94 patients in the negative group. Sinus formation (x2 = 13.028, p = 0.000), time to complete remission at the first treatment period (Z = -3.027, p = 0.002), diameter of breast mass at first-time medical consultancy (Z = -2.539, p = 0.011) and recurrence (x2 = 4.953, p = 0.026) were statistically significant. Age (Z = -1.046, p = 0.295), laterality (x2 = 4.217, p = 0.121), time to presentation since the last delivery (x2 = 0.028, p = 0.868), BMI (Z = -0.947, p = 0.344), lactation time (Z = -1.378, p = 0.168), parity (x2 = 1.799, p = 0.180), gravida (Z = -0.144, p = 0.885), history of lactational mastitis or abscess (x2 = 0.115, p = 0.734), local trauma (x2 = 0.982, p = 0.322), hyperprolactinemia (x2 = 0.706, p = 0.401), erythema nodosum (x2 = 0.292, p = 0.589), and nipple discharge (x2 = 0.281, p = 0.596) did not demonstrate statistical significance. Regarding recurrence related to therapeutic strategy, except for surgery combined with immunosuppressants (x2 = 9.110, p = 0.003), which was statistically significant, none of the other treatment regimens reached statistical significance. The recurrence rate of patients in the CK group using rifampicin in their treatment course was 22.0% (x2 = 4.892, p = 0.027). CONCLUSIONS: Granulomatous mastitis accompanied by Corynebacterium kroppenstedtii more easily forms sinuses and has a higher recurrence rate. Both of the clinical characteristics may indicate that Corynebacterium kroppenstedtii plays an important role in the development and progression of granulomatous mastitis. Lipophilic antibiotics may be essential for granulomatous mastitis with Corynebacterium kroppenstedtii infection.

Direct separation and enumeration of CTCs in viscous blood based on co-flow microchannel with tunable shear rate: a proof-of-principle study.

Circulating tumor cells (CTCs), which have extremely low density in whole blood, are an important indicator of primary tumor metastasis. Isolation and enumeration of these cells are critical for clinical applications. Separation of CTCs from massive blood cells without labeling and addition of synthetic polymers is challenging. Herein, a novel well-defined co-flow microfluidic device is presented and used to separate CTCs in viscous blood by applying both inertial and viscoelastic forces. Diluted blood without any synthetic polymer and buffer solution were used as viscoelastic fluid and Newtonian fluid, respectively, and they were co-flowed in the designed chip to form a sheath flow. The co-flow system provides the function of particle pre-focusing and creates a tunable shear rate region at the interface to adjust the migration of particles or cells from the sample solution to the buffer solution. Successful separation of CTCs from viscous blood was demonstrated and enumeration was also conducted by image recognition after separation. The statistical results indicated that a recovery rate of cancer cells greater than 87% was obtained using the developed method, which proved that the direct separation of CTCs from diluted blood can be achieved without the addition of any synthetic polymer to prepare viscoelastic fluid. This method holds great promise for the separation of cells in viscous biological fluid without either complicated channel structures or the addition of synthetic polymers.

Gradient Architecture-Enabled Capacitive Tactile Sensor with High Sensitivity and Ultrabroad Linearity Range.

The sensitivity and linearity are critical parameters that can preserve the high pressure-resolution across a wide range and simplify the signal processing process of flexible tactile sensors. Although extensive micro-structured dielectrics have been explored to improve the sensitivity of capacitive sensors, the attenuation of sensitivity with increasing pressure is yet to be fully resolved. Herein, a novel dielectric layer based on the gradient micro-dome architecture (GDA) is presented to simultaneously realize the high sensitivity and ultrabroad linearity range of capacitive sensors. The gradient micro-dome pixels with rationally collocated amount and height can effectively regulate the contact area and hence enable the linear variation in effective dielectric constant of the GDA dielectric layer under varying pressures. With systematical optimization, the sensor exhibits the high sensitivity of 0.065 kPa-1 in an ultrabroad linearity range up to 1700 kPa, which is first reported. Based on the excellent sensitivity and linearity, the high pressure-resolution can be preserved across the full scale of pressure spectrum. Therefore, potential applications such as all-round physiological signal detection in diverse scenarios, control instruction transmission with combinatorial force inputs, and convenient Morse code communication with non-overlapping capacitance signals are successfully demonstrated through a single sensor device.

Self-assembled nano-Ag/Au@Au film composite SERS substrates show high uniformity and high enhancement factor for creatinine detection.

Serum creatinine is a key biomarker for the diagnosis and monitoring of kidney disease. Rapid and sensitive creatinine detection is thus important. Here, we propose a high-performance nano-Ag/Au@Au film composite SERS substrate for the rapid detection of creatinine in human serum. Au nanoparticles (AuNPs) and Ag nanoparticles (AgNPs) with uniform particle size were synthesized by a chemical reduction method, and the nano-Ag/Au@Au film composite SERS substrate was successfully prepared via a consecutive layer-on-layer deposition using an optimized liquid-liquid interface self-assembly method. The finite element simulation analysis showed that due to the multi-dimensional plasmonic coupling effect formed between the AuNPs, AgNPs, and the Au film, the intensity of the local electromagnetic field was greatly improved, and a very high enhancement factor (EF) was obtained. Experimental results showed that the limit of detection (LOD) of this composite SERS substrate for rhodamine 6G (R6G) molecules was as low as 1 × 10-13M, and the Raman EF was 15.7 and 2.9 times that of the AuNP and AgNP monolayer substrates respectively. The results of different batch tests and SERS mapping showed that the relative standard deviations of the Raman intensity of R6G at 612 cm-1were 12.5% and 11.7%, respectively. Finally, we used the SERS substrate for the label-free detection of human serum creatinine. The results showed that the LOD of this SERS substrate for serum creatinine was 5 × 10-6M with a linear correlation coefficient of 0.96. In conclusion, the SERS substrate has high sensitivity, good uniformity, simple preparation, and has important developmental potential for the rapid detection and application of disease biomarkers.

A highly efficient preconcentration route for rapid and sensitive detection of endotoxin based on an electrochemical biosensor.

An impedimetric aptasensor for the detection of endotoxin in a microfluidic chip was proposed, in which the Apt/AuNPs/SPCE sensing surface was fabricated in a screen-printed electrode with good biological activity and stability. The quantitative detection of endotoxin was accomplished by electrochemical impedance spectroscopy (EIS) measurement before and after exposing to samples. The impedance biosensor offers an ultrasensitive and selective detection of endotoxin down to 500 pg mL-1 with a wide linear range from 500 pg mL-1 to 200 ng mL-1. According to the Langmuir isotherm model, the interactions between the target molecules and the sensing surface had been analyzed and strong binding was concluded. Compared to the traditional static incubation methods, the microfluidic biosensor realizes the enrichment of endotoxin owing to the confined space and continuous flow nature, so that the lowest detection concentration is reduced from 5 ng mL-1 to 500 pg mL-1, which is much lower than the existing technology, and the total assay time is shortened from 1.0 h to 0.5 h. The proposed microfluidic impedance biosensor provides a new strategy for the design of an aptasensor to realize the rapid detection of target biomolecules with high sensitivity and it can be integrated into wearable medical devices due to its flexible properties.

Subwavelength topological edge states based on localized spoof surface plasmonic metaparticle arrays.

Plasmonic cluster arrays have demonstrated rich physics in topological photonics, but they are seriously affected by the material loss and limited by the requirement of high-precision machining. Here, we propose a kind of ultra-thin metaparticle arrays which can mimic the coupled localized plasmonic resonances at lower frequency ranges and so that can overcome the loss and fabrication problems in real metal plasmonic systems. The metaparticle is a metallic disk with circuitous grooves that can support both spoof electric and magnetic localized resonances, and these resonances can be pushed to a subwavelength region through tuning the geometric parameters. In virtue of the highly field confinement of these localized resonances, it is thought to be an ideal experimental platform to be an analogy with various near-field interactions in topological materials. As a first proof-of-concept study to show this feasibility, the subwavelength topological edge states at the zigzag metaparticle chain boundaries are numerically and experimentally demonstrated at microwave ranges. Moreover, the subwavelength topological edge states in this zigzag chain can be excited simply by the plane wave incidence, and the edge modes at two ends can be selectively excited by controlling the polarization direction. Therefore, this kind of metaparticle array not only provides an ideal platform to experimentally study various near-filed interaction dominated topological systems but may also find massive potential applications.

Nanowire assisted repeatable DEP-SERS detection in microfluidics.

Surface enhanced Raman spectroscopy (SERS) detection in microfluidics is an interesting topic for its high sensitivity, miniaturization and online detection. In this work, a SERS detection in microfluidics with the help of the Ag nanowire aggregating based on dielectrophoresis (DEP) is reported. The Raman intensities of molecule in microfluidics is greatly enhanced in the naturally generated nanogaps of Ag nanowire aggregating modulated by DEP. Firstly, the influence of DEP voltage and time on Ag nanowire aggregating is investigated to figure out the optimal condition for SERS. And then, the SERS intensities of methylene blue and rhodamine6G at various concentration with high reproducibility and uniformity are studied. Furthermore, the experiment data demonstrate this DEP-SERS system could be repeated used for different molecule detections. At last, the SERS of melamine is measured to explore its application on food safety. Our work anticipates this nanowire assisted repeatable DEP-SERS detection in microfluidics with high sensitivity could meet the emerging needs in environmental pollution monitoring, food safety evaluation, and so on.

A fluorescent probe composed of quantum dot labeled aptamer and graphene oxide for the determination of the lipopolysaccharide endotoxin.

Endotoxins are complex lipopolysaccharides (LPS) and key components of the outer cell membrane of Gram-negative bacteria. The authors report on a fluorescent aptamer-based probe for the determination of LPS of Gram-negative bacteria. An aptamer against LPS was fluorescently labeled with CdSe/ZnS quantum dots. Its emission is quenched on addition of graphene oxide (GO). On addition of LPS, the aptamer binds LPS and GO is released. This results in the recovery of fluorescence, typically measured at excitation/emission wavelengths of 495/543 nm. The probe responds to LPS in the 10-500 ng·mL-1 concentration range, and the detection limit is 8.7 ng·mL-1. It can be used for selective detection of LPS from different Gram-negative bacteria, in the presence of biological interferents. Graphical abstract Schematic presentation of a green fluorescent probe comprised of an aptamer labelled with CdSe/ZnS quantum dots and of graphene oxide. Lipopolysaccharides bind to the aptamer and release graphene oxide to result in fluorescence recovery, which is measured at an emission wavelength 543 nm.

Dynamic Monitoring of the Oxidation Process of Phosphatidylcholine Using SERS Analysis.

Phosphatidylcholine oxidation is closely related to many neurodegenerative diseases. In this paper, Raman spectroscopy was proposed to continuously monitor the oxidation of phosphatidylcholine and provide deep understanding of this biochemical process. To increase the detection sensitivity, surface-enhanced Raman spectroscopy (SERS) with a micro-nanosilver-complex substrate was prepared by electrodeposition. The prepared SERS substrate had an enhancement factor as high as 7.8 × 107, ensuring detection sensitivity in the phosphatidylcholine-oxidation process. It was illustrated that the oxidation of phosphatidylcholine in an ethanol-water solution under the experimental conditions could be monitored and well described by second-order kinetics by continuously measuring and analyzing the SERS spectra of phosphatidylcholine-oxidation intermediates in 20 days. Meanwhile, the oxidation products were confirmed by mass spectrometry, and the oxidation process was in good concordance with mass-spectrometry detection. The use of SERS in following a biochemical process has advantages, including simple instrumentation, a low cost, a short detection time, and no sample pretreatment. Therefore, as a kind of vibration spectrum, SERS is preferable to traditional detection approaches such as MS, HPLC, and MRI for the dynamic monitoring and analysis of complex biochemical processes.

Passive Picoinjection Enables Controlled Crystallization in a Droplet Microfluidic Device.

Segmented flow microfluidic devices offer an attractive means of studying crystallization processes. However, while they are widely employed for protein crystallization, there are few examples of their use for sparingly soluble compounds due to problems with rapid device fouling and irreproducibility over longer run-times. This article presents a microfluidic device which overcomes these issues, as this is constructed around a novel design of "picoinjector" that facilitates direct injection into flowing droplets. Exploiting a Venturi junction to reduce the pressure within the droplet, it is shown that passive injection of solution from a side-capillary can be achieved in the absence of an applied electric field. The operation of this device is demonstrated for calcium carbonate, where highly reproducible results are obtained over long run-times at high supersaturations. This compares with conventional devices that use a Y-junction to achieve solution loading, where in-channel precipitation of calcium carbonate occurs even at low supersaturations. This work not only opens the door to the use of microfluidics to study the crystallization of low solubility compounds, but the simple design of a passive picoinjector will find wide utility in areas including multistep reactions and investigation of reaction dynamics.

The Effect of Additives on the Early Stages of Growth of Calcite Single Crystals.

As crystallization processes are often rapid, it can be difficult to monitor their growth mechanisms. In this study, we made use of the fact that crystallization proceeds more slowly in small volumes than in bulk solution to investigate the effects of the soluble additives Mg2+ and poly(styrene sulfonate) (PSS) on the early stages of growth of calcite crystals. Using a "Crystal Hotel" microfluidic device to provide well-defined, nanoliter volumes, we observed that calcite crystals form via an amorphous precursor phase. Surprisingly, the first calcite crystals formed are perfect rhombohedra, and the soluble additives have no influence on the morphology until the crystals reach sizes of 0.1-0.5 µm for Mg2+ and 1-2 µm for PSS. The crystals then continue to grow to develop morphologies characteristic of these additives. These results can be rationalized by considering additive binding to kink sites, which is consistent with crystal growth by a classical mechanism.

Real time hybridization studies by resonant waveguide gratings using nanopattern imaging for Single Nucleotide Polymorphism detection.

2D imaging of biochips is particularly interesting for multiplex biosensing. Resonant properties allow label-free detection using the change of refractive index at the chip surface. We demonstrate a new principle of Scanning Of Resonance on Chip by Imaging (SORCI) based on spatial profiles of nanopatterns of resonant waveguide gratings (RWGs) and its embodiment in a fluidic chip for real-time biological studies. This scheme allows multiplexing of the resonance itself by providing nanopattern sensing areas in a bioarray format. Through several chip designs we discuss resonance spatial profiles, dispersion and electric field distribution for optimal light-matter interaction with biological species of different sizes. Fluidic integration is carried out with a black anodized aluminum chamber, advantageous in term of mechanical stability, multiple uses of the chip, temperature control and low optical background. Real-time hybridization experiments are illustrated by SNP (Single Nucleotide Polymorphism) detection in gyrase A of E. coli K12, observed in evolution studies of resistance to the antibiotic ciprofloxacin. We choose a 100 base pairs (bp) DNA target (~30 kDa) including the codon of interest and demonstrate the high specificity of our technique for probes and targets with close affinity constants. This work validates the safe applicability of our unique combination of RWGs and simple instrumentation for real-time biosensing with sensitivity in buffer solution of ~10 pg/mm². Paralleling the success of RWGs sensing for cells sensing, our work opens new avenues for a large number of biological studies.

Structural dependence of silver nanowires on polyvinyl pyrrolidone (PVP) chain length.

The effect of the chain length of polyvinyl pyrrolidone (PVP) on the structures of silver nanowires (AgNWs) is explored in this study. It was found in the experiments that PVP, when serving as a capping agent, has a great impact on the morphology and structure of AgNWs. By means of a series of experiments and the inquiry of the growth mechanism, the critical minimum PVP chain length for the successful formation of uniform nanowires was discovered, below which only nanoparticles or short nanorods can be obtained. Surprisingly, a core-shell structure of a nanowire with a polycrystal was observed when PVP with a very long chain length was employed in the processing.

Fano resonance properties of gold nanocrescent arrays.

The Fano resonance induced by symmetry breaking could improve the sensitivity of localized surface plasmon resonance sensors. In this work, the spectra of gold nanocrescent arrays are measured and confirmed by simulation results through the finite element method (FEM). The Fano resonance presented in the spectra could be modulated by the symmetry breaking with different waist widths, which are understood through plasmonic hybridization theory with the help of surface charge distribution. Our results indicate the Fano lineshape is generated by the coherent coupling of the quadrupole plasmon mode QH of nanohole and the antibonding plasmon mode D(AB) of nanocrescent. Finally, the high figure of merit (FoM=1.6-3.5) of the Q mode in the visible region illustrates this nanocrescent Fano sensor is of great value in the biological and chemical scientific fields.

Gold crescent nanodisk array for nanoantenna-enhanced sensing in subwavelength areas.

Benefitting from the antenna effect and localized surface plasmon resonance (LSPR), a metal nanoparticle with a designed morphology has the amazing ability to confine light energy into the required extremely small volume, whose refractive index largely affects the optical properties of the whole metal nanoparticle. In this work, the optical spectra and near-field distribution of a gold nanocrescent array were investigated both experimentally and theoretically. To find out the LSPR wavelength and the enhancement using different morphologies of sharp tips, the spectra of gold nanocrescent arrays with different waist widths (d) were first measured, which were then confirmed and analyzed using the finite difference time-domain method and the hybridization theory. At last, the LSPR of this array with 100 nm diameter dielectric nanodisks was studied for sensing in subwavelength areas. Our results showed that because of its giant nanoantenna-enhanced electromagnetic field at the two tips, this gold nanocrescent array could be a suitable local senor to sense the variation of a local medium in a subwavelength area.

Applications of micro/nanoparticles in microfluidic sensors: a review.

This paper reviews the applications of micro/nanoparticles in microfluidics device fabrication and analytical processing. In general, researchers have focused on two properties of particles--electric behavior and magnetic behavior. The applications of micro/nanoparticles could be summarized on the chip fabrication level and on the processing level. In the fabrication of microfluidic chips (chip fabrication level), particles are good additives in polydimethylsiloxane (PDMS) to prepare conductive or magnetic composites which have wide applications in sensors, valves and actuators. On the other hand, particles could be manipulated according to their electric and magnetic properties under external electric and magnetic fields when they are travelling in microchannels (processing level). Researchers have made a great progress in preparing modified PDMS and investigating the behaviors of particles in microchannels. This article attempts to present a discussion on the basis of particles applications in microfluidics.

A pump-free paper/PDMS hybrid microfluidic chip for bacteria enrichment and fast detection.

Developing portable and sensitive biosensors for bacteria detection is highly demanded due to their association with environmental and food safety. Paper-based microfluidic chip is the suitable candidate for constructing pump-free biosensor since paper is hydrophilic, low-cost and easy to use. However, the contradiction between sensitivity and small sample volume seriously affects the application of paper-based chip for bacteria detection. Here, a new microfluidic biosensor, combining large PDMS reservoir for sample storage, hydrophilic paper substrate for pump-free water transport, coated microspheres for bacteria capture and super absorbent resin for water absorption, is designed for the detection of bacteria in aqueous samples. Once the sample solution is introduced in the reservoir, water will automatically flow through the gaps between microspheres and the target bacteria will be captured by the aptamer coated on the surface. To facilitate PDMS reservoir bonding and ensure water transport, the upper side of paper substrate is coated with Polyethylenimine modified PDMS and the bottom side is kept unchanged. After all the solution is filtrated, fluorescent dye strained bacteria are enriched on the microspheres. The fluorescent intensity representing the number of bacteria captured is then measured using a portable instrument. Through the designed microfluidic biosensor, the bacteria detection can be achieved with 2 mL sample solution in less than 15 min for water or 20 min for diluted milk. A linear range from 10 CFU/mL to 1000 CFU/mL is obtained. The paper-based 3D biosensor has the merits of low-cost, simple operation, pump-free and high sensitivity and it can be applied to the simultaneous detection of multiple bacteria via integrating different aptamers.

A bioinspired heterostructured nanochannel based on dendrimer-gold network and aluminum oxide arrays for sensitive detection of miRNA.

BACKGROUND: MicroRNAs, as oncogenes or tumor suppressors, enable to up or down-regulate gene expression during tumorigenesis. The detection of miRNAs with high sensitivity is crucial for the early diagnosis of cancer. Inspired by biological ion channels, artificial nanochannels are considered as an excellent biosensing platform with relatively high sensitivity and stability. The current nanochannel biosensors are mainly based on homogeneous membranes, and their monotonous structure and functionality limit its further development. Therefore, it is necessary to develop a heterostructured nanochannel with high ionic current rectification to achieve highly sensitive miRNA detection. RESULTS: In this work, an asymmetric heterostructured nanochannel constructed from dendrimer-gold nanoparticles network and anodic aluminum oxide are designed through an interfacial super-assembly method, which can regulate ion transport and achieve sensitive detection of target miRNA. The symmetry breaking is demonstrated to endow the heterostructured nanochannels with an outstanding ionic current rectification performance. Arising from the change of surface charges in the nanochannels triggered by DNA cascade signal amplification in solution, the proposed heterogeneous nanochannels exhibits excellent DNA-regulated ionic current response. Relying on the nucleic acid's hybridization and configuration transformation, the target miRNA-122 associated with liver cancer can be indirectly quantified with a detection limit of 1 fM and a wide dynamic range from 1 fM to 10 pM. The correlation fitting coefficient R2 of the calibration curve can reach to 0.996. The experimental results show that the method has a good recovery rate (98%-105 %) in synthetic samples. SIGNIFICANCE: This study reveals how the surface charge density of nanochannels regulate the ionic current response in the heterostructured nanochannels. The designed heterogeneous nanochannels not only possess high ionic current rectification property, but also enable to induce superior transport performance by the variation of surface chemistry. The proposed biosensor is promising for applications in early diagnosis of cancers, life science research, and single-entity electrochemical detection.

Continuous manipulation and separation of particles using combined obstacle- and curvature-induced direct current dielectrophoresis.

This paper presents a novel dielectrophoresis-based microfluidic device incorporating round hurdles within an S-shaped microchannel for continuous manipulation and separation of microparticles. Local nonuniform electric fields are generated due to the combined effects of obstacle and curvature, which in turn induce negative dielectrophoresis forces exerting on the particle that transport throughout the microchannel electrokinetically. Experiments were conducted to demonstrate the controlled trajectories of fix-sized (i.e. 10 or 15 µm) polystyrene particles, and size-dependent separation of 10 and 15 µm particles by adjusting the applied voltages at the inlet and outlets. Numerical simulations were also performed to predict the particle trajectories, which showed reasonable agreement with experimentally observed results. Compared to other microchannel designs that make use of either obstacle or curvature individually for inhomogeneous electric fields, the developed microchannel offers advantages such as improved controllability of particle motion, lower requirement of applied voltage, reduced fouling, and particle adhesion, etc.