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SARS-CoV-2, especially the variant strains, is rapidly spreading around the world. Rapid detection methods for the virus are crucial for controlling the COVID-19 epidemic. Herein, a localized surface plasmonic resonance (LSPR) biosensor based on Ω-shaped fiber optic (Ω-FO) was developed for dual assays of SARS-CoV-2 monitoring. Due to its strong ability to control the orientation and density, a new T-shaped aptamer exhibits enhanced binding affinity toward N proteins. After being combined on the fiber optic surface, the T-shaped aptamer sensitively captured N proteins of SARS-CoV-2 for a direct assay. Further, core-shell structured gold/silver nanoparticles functionalized with a T-shaped aptamer (apt-Ag@AuNPs) can amplify the signal of N protein detection for a sandwich assay. The real-time analytical feature of the dual assays endows time-dependent sensitivity enhancement behavior, which provides a guideline to save analytical time. With those characteristics, the LSPR biosensor has been successfully used to rapidly identify 39 healthy volunteers and 39 COVID-19 patients infected with the ancestral or variant SARS-CoV-2. With the help of simple pretreatment, we obtain a true negative rate of 100% and a true positive rate of 92.3% with a short analysis time of 45 min using the direct assay. Further, the LSPR biosensor could also broaden the detection application range to the surface of cold-chain foods using a sandwich assay. Thus, the LSPR biosensor based on Ω-FO was demonstrated to have broad application potential to detect SARS-CoV-2 rapidly.

Current rapid bacterial detection methods are dedicated to the classification and identification of bacteria. However, there is still a lack of a method for specific quantitative analysis of certain bacteria. In this work, a method based on elemental-tags laser-induced breakdown spectroscopy (ETLIBS) was developed for the rapid and specific quantitative analysis of Salmonella typhimurium (S. ty). Elemental tags were first synthesized by assembling copper nanoparticles (CuNPs) with poly(thymine) (poly-T) template that linked with the aptamer sequence. Under the specific recognition of the aptamer, S. ty can be fully combined with the elemental tags within 30 min to achieve labeling. Afterward, the silicon nanowires (SiNWs) array modified with Au@Ag nanoparticles (SiNWs-Au@Ag) was employed to capture S. ty in 30 min. Attributed to the rapid analysis superiority of ETLIBS mapping, 100 spectra of SiNWs-Au@Ag/S. ty/CuNPs can be obtained in 5 min. It was found that the peak area of the Cu(I) atomic emission line at 324.75 nm fitted by the Voigt profile was linearly related to the bacterial concentration in the range of 102-106 CFU/mL(R2 = 0.978). Furthermore, ETLIBS mapping achieved a low limit of detection (LOD) of 61 CFU/mL and showed good selectivity to S. ty compared with other bacteria. Besides, the method exhibited preeminent detection performance in spiked samples with the recoveries of 87-113%. With the advantages of rapidity, high efficiency, and specificity, the proposed method is expected to be a powerful tool for bacterial detection.

With cancer seriously hampering the increasing life expectancy of people, developing an instant diagnostic method has become an urgent objective. In this work, we developed a label-free laser-induced breakdown spectroscopy (LIBS) method for high-throughput recognition of tumor cells. LIBS spectra were straightly collected from cells dropped on a silicon substrate and built into a deep learning model for simultaneous classification of various cancers. To interpret the result of the deep learning algorithm, gradient-weighted class activation mapping was utilized to a one-dimensional convolution neural network (1D-CNN), and the saliency maps thus obtained amplified the differences between the spectra of cell lines. Overall results showed that the 1D-CNN algorithms achieved a mean sensitivity of 94.00%, a mean specificity of 98.47%, and a mean accuracy of 97.56%. Thus, the proposed method performed satisfactorily and is seen as an interpretable classification process for cancer cell lines.

Signal amplification is critical to achieving sensitive biosensing, but complex strategies often bring problems like system instability, false positive, or narrow target spectrum. Here, a self-extending DNA-mediated isothermal amplification (SEIA) system with simple reaction components is introduced to achieve rapid, robust, and significant signal amplification. In SEIA, based on spontaneous refolding of specific DNA domains and using the previous generation product as a template, a DNA strand can extend continuously in an approximate exponential growth pattern, which was accurately predicted by our formula and well supported by AFM results. Based on a set of proof-of-concept experiments, it was proved that the SEIA system can output different signals and flexibly integrate various functional nucleic acids, which makes it suitable for different scenarios and realizes broad-spectrum target detection. Taking into account the advantages of simplicity, flexibility, and efficiency, the SEIA system as an independent signal amplification module will enrich the toolbox of biosensing design.

A label-free and enzyme-free colorimetric sensor for rapid detection of Pb2+ is reported, which is based on the strategy of DNAzyme-mediated RNA cleavage combined with an annealing-accelerated DNA hybridization chain reaction (HCR). As a trigger DNA, the substrate strand (STM) of DNAzyme can initiate HCR effectively. However, when it is cleaved by DNAzyme in the presence of Pb2+, the separation of DNA functional domains leads to a serious decrease in HCR efficiency. As a result, the difference in Pb2+ concentration converts into the difference of DNA assembly, which eventually leads to the color change of colloidal gold nanoparticles (AuNPs). In this work, a DNA strand (cGR5) completely complementary to the catalytic strand (GR5) of DNAzyme is used to improve the dissociation of STM to enhance the HCR efficiency. In addition, the simple operation of DNA annealing is first used to accelerate the HCR process, enabling the Pb2+ detection to be completed in about 30 min. As advantages of high sensitivity, goo...

Ultrasensitive detection of metallic elements in liquids has attracted considerable attention in fields such as environmental pollution monitoring and drinking water quality control. Hence, it is of great significance to develop a sensitive and simultaneous detection strategy for multiple metal elements in liquid. Laser-induced breakdown spectroscopy (LIBS) technology shows unique advantages because of its simple, rapid, and real-time in situ detection, but the laser energy will be greatly attenuated in the liquids; thus, the sensitivity of LIBS for direct detection of metal elements in liquid samples will decrease sharply. In this study, inspired by the structure of Stenocara beetle's back, a superhydrophobic biomimetic interface with hydrophilic array was prepared for enriching low-concentration targets into detection regions, and the biomimetic array LIBS (BA-LIBS) was successfully established. The ultrasensitive and simultaneous detection of nine metal elements in drinking water was realized based on the effective enrichment method. The limits of detection of the nine metal elements in mixed solution ranged from 8.3 ppt to 13.49 ppb. With these excellent properties, this facile and ultrasensitive BA-LIBS strategy might provide a new idea for the prevention and control of metal hazards in the liquid environment.

Current rapid bacterial detection methods are dedicated to the classification and identification of bacteria. However, there is still a lack of a method for specific quantitative analysis of certain bacteria. In this work, a method based on elemental-tags laser-induced breakdown spectroscopy (ETLIBS) was developed for the rapid and specific quantitative analysis of

A novel, Ω-shaped fiber-optic localized surface plasmon resonance (FOLSPR) biosensor was designed for sensitive real-time and label-free bacterial detection. The designed Ω-shaped fiber-optic probe exhibits an outstanding sensitivity, due to the effect of unique geometry on performance. The results show that refractive index (RI) sensitivity of the Ω-shaped fiber-optic probe is 14 times and 2.5 times higher than those of the straight-shaped and the U-shaped FOLSPR, respectively. In addition, the reason for the geometry and the bending radius effects on RI sensitivity was discussed by investigating the relationship between RI sensitivity and the bending area. The results show that RI sensitivity was enhanced with the increase of bending area, and the best RI sensitivity obtained by Ω-shaped FOLSPR was 64.582 (a.u.)/RIU. Combined with this newly designed Ω-shaped FOLSPR biosensor, a real-time, label-free, sensitive, and highly selective bacterial detection method was established. In this work, the aptamers immobilized on the surface of FOLSPR could specifically capture Salmonella Typhimurium, resulting in an intense change of the absorption peak. In line with this principle, the FOLSPR biosensor achieved high detection sensitivity for Salmonella Typhimurium down to 128 CFU/mL within a linear range from 5 × 10

The threat of food safety and the limited analytical methods with high performance promote the growing interest in the development of pathogenic bacteria biosensors. This study presents a pathogenic bacteria biosensing system, where a novel three-dimensional (3D) chip acts as an analytical carrier and DNA-programmed hybridization chain reaction (HCR) causes signal amplification. The 3D chip is designed featuring a compact multichannel structure. It has a large surface area for sensitive sensing and exhibits multiple functions of target capture, separation, rinsing, and signal detection to simplify the analysis processes. HCR, which enables the fluorophore's polymerization, is designed as two signal amplification modes, each with unique advantages. Mode I achieves highly sensitive detection in a "sandwich" assay format, in which a long HCR-amplified probe is used to boost the fluorescence signal. In mode II, the assembly of HCR is performed on the inner surface of the 3D chip. Especially, a group of rapid-assembly HCR sequences is proposed, of which the assembly time as short as 15 min stands out among the related works previously reported. Under the optimal conditions, the proposed biosensing system has the limits of detection (LOD) of 4 and 8 cfu/mL in mode I for Staphylococcus aureus detection and in mode II for Salmonella enterica Typhimurium detection, respectively. The specificity and the real sample applications are evaluated. This multichannel-structured 3D chip based on HCR signal amplification has potential applications in food safety monitoring and biosensor development.

A label-free and enzyme-free colorimetric sensor for rapid detection of Pb

To expand the applications of glow discharge microplasma into biological analysis, an innovative ambient ion source for mass spectrometry, microplasma probe desorption/ionization mass spectrometry (MPPDI-MS), has been developed and demonstrated. Electrodes and a sampling tube were creatively combined using a stainless steel syringe needle, and efficient methods of introduction for biological samples in solid, liquid, and gaseous phases like phospholipid and amino acids were specially designed. Based on the active species generated by glow discharge plasma, simplified protonated spectra were obtained without extra solvent spray assistance. The method is easy to operate and versatile and especially has the ability to distinguish the isomeric compounds of ketone and aldehyde. Quantitative results of this method for different biological samples in different phases were also performed well. It was proved that with further improvement, this sensitive and selective analysis using MPPDI-MS with minimal invasiveness will be an ingenious tool in disease diagnosis and single-cell detections in the future.

In this study, nitrogen was successfully used to maintain the microplasma discharge to excite and detect organic compounds for the first time. A new nitrogen glow discharge microplasma-generated in-chip-based ingroove device was developed and applied as the excitation source for optical emission spectrometry. The unique ingroove design of the discharge chamber can provide good stability and sensitivity for nitrogen microplasma to detect trace organic samples. Unlike argon/helium microplasmas, the nitrogen microplasma has a strict demand on the material of electrodes, especially cathodes. We tested the effects of four common electrode materials from various aspects and obtained the most appropriate material for nitrogen glow discharge, namely, platinum. We also studied the excitation and detection mechanism of organic compounds in nitrogen microplasma and confirmed that the main means of excitation in nitrogen plasma was through energy transfer rather than penning ionization. Several organic compounds were directly injected and detected in the optimized working conditions with the limits of detection at the hundreds of picograms level. Because of the portable nitrogen generators handily and commercially available, this detection system with nitrogen as the discharge gas can overcome the limitation of noble gas supply. In addition, the advantages of small size, low energy consumption, good stability, and reproducibility demonstrate that the nitrogen ingroove microplasma source can be applied in a portable detector for on-site and real-time spectrometry detection.

A novel ambient ionization technique for mass spectrometry, microfabricated glow discharge plasma (MFGDP), is reported. This device is made of a millimeter-sized ceramic cavity with two platinum electrodes positioned face-to-face. He or Ar plasma can be generated by a direct current voltage of several hundreds of volts requiring a total power below 4 W. The thermal plume temperature of the He plasma was measured and found to be between 25 and 80 °C at a normal discharge current. Gaseous, liquid, creamy, and solid samples with molecular weights up to 1.5 kDa could be examined in both positive and negative mode, giving limits of detection (LOD) at or below the fg/mm(2) level. The relative standard deviation (RSD) of manual sampling ranged from 10% to ~20%, while correlation coefficients of the working curve (R(2)) are all above 0.98 with the addition of internal standards. The ionization mechanisms are examed via both optical and mass spectrometry. Due to the low temperature characteristics of the microplasma, nonthermal momentum desorption is considered to dominate the desorption process.

A fast, low cost, and sensitive sample pretreatment method was specially developed for laser-induced breakdown spectroscopy (LIBS) based on metal precipitation and membrane separation for simultaneous elemental analysis in liquid samples. The metal elements were reacted with the chelating reagent 2,4,6-trimercapto-1,3,5-triazine (TMT) in the first step and separated by mixed cellulose ester microfiltration membrane subsequently. A specific membrane supporter with smaller aperture than the commercially available needle filter was designed and assembled in the presented research to increase the sensitivity. As a result, the detection limits of Cu, Ag, Mn, and Cr obtained in this research were 2.59 ng·mL(-1), 0.957 ng·mL(-1), 0.958 ng·mL(-1), and 1.29 ng·mL(-1) respectively, which were greatly improved from direct liquid analysis by LIBS. Satisfactory linearity, reproducibility, and accuracy were also obtained in the concentration range of 10-120 ng·mL(-1) for all four elements tested at the optimized experimental conditions. The analytical figures of merits of the proposed method are at an advanced level and are comparable to those reported among other non-LIBS research. In addition, the separation mechanism in this research was initially explored and further varied to be based on metal precipitates adsorption by membrane fibers, rather than a regular size-dependent obstruction.

An innovative signal-enhancing immunoassay for ultrasensitive biomarker detection based on laser-induced fluorescence (LIF) has been developed. A novel LIF optical system with high collection efficiency was constructed by using a parabolic mirror. Carboxyl-functionalized magnetic beads were used to immobilize antibody for achieving a conventional sandwich assay. Fluorescence from Rhodamine 6G (R6G)-labeled antibody was collected by the newly designed optical system. To reduce photobleaching of R6G under laser irradiation, ethanol instead of commonly used aqueous solution was used as assay buffer in the last stage. The newly developed LIF immunoassay displayed excellent analytical performance for α-fetoprotein (AFP) detection in the concentration range from 0.005 to 1.0 ng/mL with a detection limit of 0.0016 ng/mL. The detection limit obtained in this work is about 3 orders of magnitude better than that of conventional enzyme-linked immunosorbent assay (ELISA). In addition, the proposed method exhibited excellent precision, acceptable stability, and good reproducibility. Furthermore, the proposed immunoassay was successfully applied to AFP determination in real serum specimens. Therefore, the present immunoassay was demonstrated to be a powerful tool for ultrasensitive biomarker detection.

We have been exploring innovative technologies for elemental and hyperfine structure measurements using cavity ring-down spectroscopy (CRDS) combined with various plasma sources. A laboratory CRDS system utilizing a tunable dye laser is employed in this work to demonstrate the feasibility of the technology. An in-house fabricated sampling system is used to generate aerosols from solution samples and introduce the aerosols into the plasma source. The ring-down signals are monitored using a photomultiplier tube and recorded using a digital oscilloscope interfaced to a computer. Several microwave plasma discharge devices are tested for mercury CRDS measurement. Various discharge tubes have been designed and tested to reduce background interference and increase the sample path length while still controlling turbulence generated from the plasma gas flow. Significant background reduction has been achieved with the implementation of the newly designed tube-shaped plasma devices, which has resulted in a detection limit of 0.4 ng/mL for mercury with the plasma source CRDS. The calibration curves obtained in this work readily show that linearity over 2 orders of magnitude can be obtained with plasma-CRDS for mercury detection. In this work, the hyperfine structure of mercury at the experimental plasma temperatures is clearly identified. We expect that plasma source cavity ring-down spectroscopy will provide enhanced capabilities for elemental and isotopic measurements.

We are exploring sensitive techniques for elemental measurements using cavity ring-down spectroscopy (CRDS) combined with a compact microwave plasma source as an atomic absorption cell. The research work marries the high sensitivity of CRDS with a low-power microwave plasma source to develop a new instrument that yields high sensitivity and capability for elemental measurements. CRDS can provide orders of magnitude improvement in sensitivity over conventional absorption techniques. Additional benefit is gained from a compact microwave plasma source that possesses the advantages of low power and low-plasma gas flow rate, which are of benefit for atomic absorption measurements. A laboratory CRDS system consisting of a tunable dye laser is used in this work for developing a scientific base and demonstrating the feasibility of the technique. A laboratory-designed and -built sampling system for solution sample introduction is used for testing. The ring-down signals are monitored using a photomultiplier tube and recorded using a digital oscilloscope interfaced to a computer. Lead is chosen as a typical element for the system optimization and characterization. The effects of baseline noise from the plasma source are reported. A detection limit of 0.8 ppb (10(-)(10)) is obtained with such a device.

A low-power, plasma source-based, portable molecular emission detector is described in this paper. The detector employs a pulsed-plasma source operated at atmospheric pressure for molecular fragmentation and excitation. The plasma was generated with a home-built high-voltage pulsed power supply. The average operational power of the detector was less than 0.2 W. The effects of operational parameters such as plasma gas, voltage, and plasma gas flow rate were investigated. Molecular emission spectra of a variety of organic compounds were studied. The features of the emission spectra obtained with the pulsed plasma source were significantly different from those obtained with direct current (dc) discharge at a power higher than 10 W. The spectra obtained in this work showed strong CH emission at 431.2 nm; however, the typical CN emission observed with a conventional dc plasma source at 383-388 nm was very weak in most cases. The strong CN emission was only obtained for compounds containing nitrogen, such as aniline. Dimethyl sulfoxide can be detected at a limit of 200 ppb using helium plasma by observing the emission band of the CH radical. The detector was very stable and did not experience electrode fouling even with the introduction of organic vapors. Such a detector is very promising for organic vapor detection.

A novel capillary-based three-dimensional (3D) fluoroimmunosensor for carcinoembryonic antigen detection was explored for the first time. The immunosensor was designed in symmetrical cylinder structure and fabricated with capillary tubes encapsulated in a quartz tube. The 3D design of the sensor increased the area of sensing surface, flexibility in light path design and efficiency of fluorescence collection by aluminum foil, resulting in analytical performance improvement. The CEA immunosensor was constructed in double antibody sandwich format. Fluorescence signals from DyLight 550-labeled antibody were measured using a laser-induced fluorescence spectrometry. There is an obvious improvement in the linear detection range of 0.7-80 ng/mL. This novel 3D immunosensor dramatically improved the detection limit (1.1 pmol/L CEA) and sensitivity. Assay validation studies indicated that the correlation coefficient reached 0.9935, with recoveries of 92.82-118.81%. Furthermore, the immunosensor was successfully applied to CEA determination in actual saliva specimens with high sensitivity and acceptable precision. Regarding accuracy, the results obtained by 3D immunosensor were not significantly different (t test) from those obtained by validated electrochemiluminescence immunoassay. This new 3D CEA immunosensor was demonstrated to be a high-performance tool for CEA diagnostics.

In this study, antibody immobilization using a microwave-induced H2O/Ar plasma pretreatment was achieved for the first time. Plasma was used to activate the surface of a capillary-based immunosensor by increasing the density of silicon hydroxyls and dangling bonds to ensure better silanization. The capture antibodies were covalently immobilized after the silanized surface reacted with glutaraldehyde and antibodies. A Cy3-labeled detection antibody was used in combination with the antigen captured by the immunosensor to complete the sandwich-type immunoassay, and the signals were measured using a laser-induced fluorescence system. Microwave-induced H2O/Ar plasma pretreatment of the carcinoembryonic antigen (CEA) immunosensor improved the antibody immobilization, and there was an obvious improvement in the linear detection range, i.e., 1 order of magnitude compared with a commercial enzyme-linked immunosorbent assay (ELISA). This novel immobilization method dramatically improved the detection limit (0.5 pmol/L CEA) and sensitivity. Assay validation studies indicated that the correlation coefficient reached 0.9978, and the relative standard deviations were7% for all samples, with recoveries of 99.7-107.1%. Furthermore, the immunosensor was applied successfully to CEA determination in actual saliva specimens with high sensitivity, acceptable precision, and reasonable accuracy. This enhanced CEA immunosensor based on microwave-induced H2O/Ar plasma was demonstrated to be a sensitive tool for CEA diagnostics.

A new ionization source based on microwave induced plasma was developed for ambient desorption/ionization. The microwave-induced plasma desorption/ionization source (MIPDI) was composed of a copper Surfatron microwave cavity where a fused-silica tube was centered axially. Stable nonlocal thermodynamic equilibrium plasma was generated in the quartz discharge tube when a microwave at a frequency of 2450 MHz was coupled to the microwave cavity. Analytes deposited on the surface of poly(tetrafluoroethylene) (PTFE) or quartz slide after hydrofluoric acid (HF) etching were desorbed and ionized by the plasma. The performance of the MIPDI technique was validated by the analysis of a variety of chemical substances, polymer compounds, and pharmaceutical drugs using argon or helium as the discharge gas. Protonated [M + H](+) or deprotonated [M - H](-) ions were observed in the positive or negative mode. MIPDI was also used for the analysis of compounds in a complex matrix without any sample preparation. MIPDI was also capable of analyzing liquid samples. The signal-to-noise ratio was 463 in the analysis of 9.2 ng of phenylalanine, and the limit of detection was 60 pg for phenylalanine. MIPDI could desorb and ionize analytes with a molecular weight of up to 1200, which was demonstrated by the analysis of polyethylene glycol 800 (PEG800). MIPDI has advantages of simple instrumentation, relatively high temperature, stability, and reproducibility.

In this paper, an atmospheric pressure, helium microwave-induced plasma (MIP) ion source coupled with an orthogonal acceleration time-of-flight mass spectrometer (TOFMS) is explored for elemental analysis. Studies of the relationship between ion signals and sampling distance of the MS reveal that background signals can be suppressed dramatically without sacrificing the signal intensities of analytes when the microwave plasma plume is off the tip of the sampler orifice. This "off-cone" ion sampling mode provides a technique to obtain nearly "clean" background spectra and, thus, eliminates the spectral interference from entrainment air and the working-gas species, making it possible to sensitively determine isotopes that suffered from spectral interference in ICPMS and MIPMS (such as 40Ca, 52Cr, 55Mn, and 56Fe). On the other hand, since the high-temperature plasma is kept away from the sampler aperture, off-cone sampling places little demand on the cooling device and the lifetime of the sampler plate can be extended. The instrumental system can provide a fairly good mass resolution of 1100 (fwhm). The detection limits (3sigma) in the tens of picograms per milliliter level for the elements studied can be achieved with a digital oscilloscope. These detection limits can be easily improved with an advanced detection system, which is currently available in commercial markets.

In this research, a novel portable instrument for on-site, real-time air particulate monitoring was developed. It is highly desirable to use microwave plasmas for on-site, real-time environmental and occupational hazard monitoring because they can be sustained with various gases at relatively low power and possess excellent detection capabilities for both metal and nonmetal air pollutants. In the new instrument design, a microwave plasma was selected as an excitation source and was used in conjunction with atomic emission spectrometry. A small, integrated spectrometer with a charge-coupled detector (CCD) was used for optical signal detection. An efficient, in situ air-sampling system was developed for direct sampling of air particles into the plasma. Characterization and calibration of the new instrument were achieved with an in-house-fabricated high-efficiency nebulization-desolvation system. Tolerance of the microwave argon plasma source to air introduction was tested, and the operational parameters were optimized. Analytical performance and the feasibility of the newly developed portable instrument for aerosol particle analysis were evaluated. Some advantages and possible applications of the new instrument are discussed. The instrument provides an innovative tool for rapid environmental and occupational hazard monitoring.

In this work, a new glow discharge, microwave-induced-plasma (GDMIP), tandem ion source was developed, characterized, and used in conjunction with time-of-flight mass spectrometry. This tandem source was designed to be simple and compact The GD and the MIP unit are independent and demountable, providing flexibility in tuning and operating. The microwave plasma can be formed with very low power (a few watts) and overlapped with the glow discharge. The discharge current increased with the addition of microwave discharge when the GD was operated in constant-voltage mode, and the amplitude of the current increase was found to be related to microwave power, discharge pressure, and sampling distance. With the addition of microwave discharge, significant signal enhancement was achieved under a discharge pressure of 2.0 Torr and a discharge voltage of 500 V with 2.0-mm sampling distance. Enhanced factors of analyte signals ranging from approximately 1 to nearly 10 could be obtained with MIP boosting, while no significant change in noise level was observed. This new tandem source provides improved performance in direct solid sample analysis.