The growing use of aptamers as target recognition elements in label-free biosensing necessitates corresponding transducers that can be used in relevant environments. While popular in many fields, capacitive sensors have seen relatively little, but growing use in conjunction with aptamers for sensing diverse targets. Few reports have shown physiologically relevant sensitivity in laboratory conditions and a cohesive picture on how target capture modifies the measured capacitance has been lacking. In this review, we assess the current state of the field in three areas: small molecule, protein, and cell sensing. We critically analyze the proposed hypotheses on how aptamer-target capture modifies the capacitance, as many mechanistic postulations appear to conflict between published works. As the field matures, we encourage future works to investigate individual aptamer-target interactions and to interrogate the physical mechanisms leading to measured changes in capacitance. To this point, we provide recommendations on best practices for developing aptasensors with a particular focus on considerations for biosensing in clinical settings.

DNAzyme-based electrochemical biosensors provide exceptional analytical sensitivity and high target recognition specificity for disease diagnosis. This review provides a critical perspective on the fundamental and applied impact of incorporating DNAzymes in the field of electrochemical biosensing. Specifically, we highlight recent advances in creating DNAzyme-based electrochemical biosensors for diagnosing infectious diseases, cancer and regulatory diseases. We also develop an understanding of challenges around translating the research in the field of DNAzyme-based electrochemical biosensors from labs to clinics, followed by a discussion on different strategies that can be applied to enhance the performance of the currently existing technologies to create truly point-of-care electrochemical DNAzyme biosensors.

Advanced biosensors, considered as emerging technologies, are capable of accurate, quantitative and real-time analysis for point-of-care testing (POCT) applications. Moreover, the integrating of miniaturized meters into these advanced biosensors makes them ideally appropriate for portable, sensitive and selective detection of biomolecules. Miniaturized meters including PGMs (personal glucose meters), thermometer, pressuremeter, pH meter, etc. are the most accurate devices and wide availability in the market, exhibiting a promising potential towards detection of small molecule mycotoxins. In this article, we introduce and analyze the recent advancements for sensing of mycotoxins measured by handheld meters since the first report in 2012. Furthermore, limitations and challenges for versatile meters application against mycotoxins in food matrix are highlighted. By overcoming the bottleneck problems, we believe the miniaturized meters-based biosensor platform will provide great possibilities for mycotoxins analysis and launch them to the market.

MiRNAs hold great potential as biomarkers for the early detection and monitoring of diseases based on their differential expression profiles. Therefore, the sensitive, specific and accurate detection of miRNAs represents an emerging new tool to improve diagnosis and treatment of several diseases, cancer in particular. DNA origami-based miRNA detection is particularly advantageous as it allows to incorporate multiple attachment sites to capture different target miRNAs at the nanoscale. In this work, we present a DNA origami nanoarray system providing distance-dependent recognition of miRNAs by applying super-resolution microscopy technique; DNA-PAINT (point accumulation for imaging in nanoscale topography). The sensor can detect up to 4 miRNAs either separately or in combination based on the relative distance to the boundary markers on the structure using a single imager strand. The detection is highly sensitive, with a limit of detection down to the low femtomolar range (11 fM - 388 fM) and has a large dynamic range up to 10 nM without need for amplification. Moreover, our detection system can discriminate single base mismatches with low false positive rates. Using our strategy, we demonstrate the detection of endogenous miRNAs from cell extracts of cancer cell lines and plasma from breast cancer patients. Overall, we developed an ultrasensitive and amplification-free, DNA-PAINT imaging-based miRNA detection method using DNA origami nanoarray system for the detection of breast-cancer associated miRNAs which potentially provides a sensitive and accurate alternative to the current multiplexed diagnostic technologies.

The human body detects tactile stimuli through a combination of pressure force and temperature signals via various cutaneous receptors. The development of a multifunctional artificial tactile perception system has potential benefits for future robotic technologies, human-machine interfaces, artificial intelligence, and health monitoring devices. However, constructing systems beyond simple pressure sensing capabilities remains challenging. Here, we propose an artificial flexible and ultra-thin (50 μ m) skin system to simultaneously capture 3D tactile and thermal signals, which mimics the human tactile recognition process using customized sensor pairs and compact peripheral signal-converting circuits. The 3D tactile sensors have a flower-like asymmetric structure with 5-ports and 4 capacitive elements in pairs. Differential and average signals would reveal the curl and amplitude values of the fore field with a resolution of 0.18/mm. The resistive thermal sensors are fabricated with serpentine lines and possess stable heat-sensing performance (165 mV/°C) under shape deformation conditions. Real-time monitoring of the skin stimuli is displayed on the user interface and stored on mobile clients. This work offers broad capabilities relevant to practical applications ranging from assistant prosthetics to artificial electronic skins.

Fluorescent lateral flow immunoassay (LFA) is one of the most common analytical platforms for point-of-care testing (POCT), which is capable of facile and early screening of biomarkers. Notably, fluorescent probes play a decisive role in analytical performances of LFA. Herein, we report a novel LFA based on the rare earth doped nanoparticles (RENPs) emitting in the second near-infrared (NIR-II) window for the detection of biomarkers, such as carcinoembryonic antigen (CEA). Benefiting from the dual fluorescent emission at NIR-II window, strong fluorescent penetration, low autofluorescence and excellent photostability of RENPs, this proposed NIR-II LFA displays a good linear relationship ranging from 1 to 320 ng mL-1. The detection limit is as low as 0.37 ng mL-1, which is of 13.5 times lower than the clinical cutoff value. Overall, NIR-II LFA biosensing platform based RENPs not only exhibits high sensitivity, accuracy and specificity, but also have characteristics of rapidity, simplicity and low cost. It holds high potential for early diagnosis of tumor biomarkers in POCT.

Gastric acid is an important functional substance secreted by the stomach of the living organisms, reflecting the gastric physiological condition. The sensing of gastric acid in vivo is of great significance for evaluation of gastric function, diagnosis and treatment of gastric diseases and maintenance of organism health but remains challenging due to the harsh acid and digestive environment of stomach. This study developed an activatable nanoprobe based on Au nanoclusters (Au NCs) for sensitive and real-time noninvasive near-infrared II (NIR-II) fluorescence imaging detection of gastric acid in vivo for the first time. The Au NCs were encapsulated by polydopamine to have enhanced NIR-II luminescence and high stability and combined with methylene blue to possess the pH responsiveness for gastric acid imaging. The developed nanoprobe could not only monitor gastric acid secretion in vivo but also imaging the changes of gastric acid caused by feeding, acid-inhibition drugs and gastric ulcer disease. This study provides a promising avenue for the improvement of the application performance of Au NCs and imaging analysis of gastric acid and related gastric diseases.

Amide hydrolysis using enzyme labels, such as proteases, occurs at a slower rate than phosphoester and carboxyl ester hydrolysis. Thus, it is not very useful for obtaining high signal amplification in biosensors. However, amide hydrolysis is less sensitive to nonenzymatic spontaneous hydrolysis, allowing for lower background levels. Herein, we report that amide hydrolysis by DT-diaphorase (DT-D) occurs rapidly and that its combination with five redox-cycling reactions allows for the development of a highly sensitive electrochemical immunosensor. DT-D rapidly generates ortho-aminohydroxy-naphthalene (oAN) from its amide substrate via amide hydrolysis, which not even trypsin, a highly catalytic protease, can achieve. NADH, which is required for amide hydrolysis, advantageously acts as a reducing agent for rapid electrooxidation-based redox-cycling reactions. In the presence of oAN, DT-D, and NADH, two redox-cycling reactions rapidly occur. In the additional presence of an electron mediator, Ru(NH3)63+ [Ru(III)], three more redox-cycling reactions occur because Ru(III) reacts rapidly with oAN and DT-D. Although the O2-related redox-cycling reactions and redox reaction decrease electrochemical signals, this signal-decreasing effect is not significant in air-saturated solutions. The slow electrooxidation of NADH at an indium tin oxide electrode and sluggish reaction between NADH and Ru(III) allow for low electrochemical backgrounds. When the developed signal amplification scheme is tested for the sandwich-type electrochemical detection of parathyroid hormone (PTH), a detection limit of ∼1 pg/mL is obtained. The detection method is highly sensitive and can accurately measure PTH in serum samples.

In this study, based on the oxidase activity and photothermal effect of manganese dioxide nanosheets (MnO2 NSs), with thiamine (TH) as the fluorescence response signal and tris (2,2'-bipyridyl) ruthenium (II) hexahydrate as the reference signal, an enzyme-regulated ratiometric fluorescence and photothermal dual-mode probe was constructed for the quantitative detection of organophosphorus pesticide (OPs) residues. OPs reduced the production of the reductive product thiocholine by inhibiting the activity of acetylcholinesterase, thereby regulating the residual amount of MnO2 NSs. With the increase of OPs concentration, the color of the probe solution gradually transitioned from red to blue, and the temperature gradually increased. Using dichlorvos and chlorpyrifos as pesticide models, the developed probes exhibited sensitive responses to OPs in a wide linear range of 0.1-8000 ng/mL. The detection limits of dichlorvos and chlorpyrifos in fluorescence mode were 1.13 × 10-3 ng/mL and 0.86 ng/mL, respectively. The corresponding detection limits in photothermal mode were 1.01 ng/mL and 1.02 ng/mL, respectively. The proposed probe displayed excellent anti-interference and reliability in the analysis of OPs residues in real samples. The dual-mode probe with self-verification function is expected to provide more accurate and robust detection results than the single-mode probe, and has a wider application prospect.

Accurate quantitative, in situ and temporal tracking imaging of tumor-associated miRNAs in living cells could provide a basis for cancer diagnosis and prognosis. In this strategy, a surface-enhanced Raman scattering (SERS)-fluorescence (FL) dual-spectral sensor (DSS) was constructed based on the nanoscale photophysical properties of AuNPs, mediated by functionalized DNA, to achieve rapid imaging of FL and accurate SERS quantification of intracellular miRNAs. The dual-spectrum sensor in the strategy is highly sensitive, specific and reproducibly stable. The LOD values of the dual spectra were 3.58 pM (SERS) as well as 11.8 pM (FL) with RSD values less than 2.69%. The bispectral sensor self-assembled into a trimer by the lapidation of Y-type DNA under the excitation of the target, generating a stable enhanced electric field coupling; and selected adenine located in the enhanced electric field as the reporter molecule, simplifying the labeling process and variables of the Raman reporter molecule, distinguishing it from other traditional methods. This strategy successfully achieved accurate tracking and quantification of miR-21 in cancer cells and showed good stability in the cells. The reported probes are potential tools for reliable monitoring of biomolecular dynamics in living cells.

The rapid, low-cost and user-friendly methods for nucleic acid detection is urgently needed. Here we developed a miniaturized and convenient photoelectrochemical biosensor array (PEBA) platform for multiplexing and simultaneous detection of nucleic acid. The array system containing nine sensing units was integrated on one piece of conductive glass by laser etching and screen printing. Moreover, human papillomavirus (HPV), the main cause of cervical cancer, was selected as the model marker to evaluate the applicability of the fabricated PEBA. The proposed PEBA for HPV genotyping involved the TiO2@Au nanoparticles (NPs) as the photoelectrochemical (PEC) material and CdS quantum dots (CdS QDs)-labeled DNA hairpin probe anchored on the TiO2@Au NPs as the recognition element. With the addition of HPV target, the probe undergoes a significant conformational change and forces CdS QDs away from TiO2@Au, resulting in decreased PEC signals. The established array platform coupled with nucleic acid amplification exhibited high sensitivity as low as 0.1 copies/μL and a linear range of 0.6-600 copies/μL for nine HPV genotyping. Method evaluation with 40 clinical samples including 20 HPV-positive and 20 negative samples, gave a 97.5% concordance result in comparison with commercial kits. The genotyping results obtained by the PEBA reveal that HPV52, HPV18, and HPV11 are the most prevalent genotypes in positive samples, which is in good concordance with the official report concerning the trend of HPV prevalence among women with cervical lesions in Shenzhen. The designed PEBA platform has potential applications in extensive fields like biomedicine analysis and clinical healthcare diagnosis.

Toxicity screening and risk assessment of an overwhelmingly large and ever-increasing number of chemicals are vitally essential for ecological safety and human health. Genotoxicity is particularly important because of its association with mutagenicity, carcinogenicity and cancer. Phosphorylated histone H2AX (γH2AX) is an early sensitive genotoxic biomarker. It is therefore highly desirable to develop analytical methods for the detection of trace γH2AX to enable screening and assessment of genotoxicity. Here, we developed a novel cathodic photoelectrochemical (PEC) immunoassay with dual signal amplification for the rapid and ultrasensitive detection of γH2AX in cell lysates. A sandwich immuno-reaction targeting γH2AX was first carried out on a 96-well plate, using a secondary antibody/gold nanoparticle/glucose oxidase conjugate as the labeled detection antibody. The conjugate increased the production of H2O2 and thus provided the first mechanism of signal amplification. The immuno-reaction product containing H2O2 was then detected on a photocathode prepared from Bi2+xWO6 rich in oxygen vacancies, with H2O2 acting as electron acceptor. The oxygen vacancies acted as both adsorption and activation sites of H2O2 and thus enhanced the photocurrent, which provided another mechanism of signal amplification. As a result, an ultrasensitive immunoassay for γH2AX determination was established with a limit of detection of 6.87 pg/mL (S/N = 3) and a wide linear range from 0.01 to 500 ng/mL. The practicability of this assay was verified by detecting γH2AX in cell lysates exposed to known genotoxic chemicals. Our work offers a promising tool for the screening of genotoxic chemicals and opening a new avenue toward environmental risk assessment.

Current single signal electrochemiluminescence (ECL) sensors are susceptible to false positive or false negative phenomena due to experimental conditions. Therefore, sensors with "self-checking" function are attracting democratic attention. In quick succession, a highly sensitive single-cathode dual ECL signal aptasensor with self-checking function to improve the shortcomings mentioned above was designed. This aptasensor used In-based metal-organic framework (MIL-68) as load and stabilizer to effectively attenuate the aggregation-induced quenching (ACQ) effect of porphyrin derivatives (Sn-TCPP) while improve ECL stability. The introduction of cooperative-binding split-aptamers" (CBSAs) aptamers increased the specificity of the aptasensor and its unique double-binding domains detection accelerated the detection efficiency. When analyzing 3,4-methylenedioxypyrovalerone (MDPV), we could calculate two concentrations based on the strength of ECL 1 and ECL 2. If the concentrations are the same, the result would be obtained; if not, it should be retested. Depending on the above operation, the results achieve self-check. It was found that the designed aptasensor could quantify the concentration of MDPV between 1.0 × 10-12 g/L and 1.0 × 10-6 g/L with the limit of detection (LOD) of 1.4 × 10-13 g/L and 2.0 × 10-13 g/L, respectively (3 σ/slope). This study not only improves the detection technology of MDPV, but also explores the dual-signal detection of porphyrin for the first time and enriches the definition of self-checking sensor.

Electrochemical sensors are widely used for nucleic acid detection. However, they exhibit low sensitivity and specificity. To overcome these limitations, DNA amplification method is necessary. In this study, we introduced CRISPR (Clustered regularly interspaced short palindromic repeats) Cas12a-dependent hyperbranched rolling circle amplification (HRCA) into an electrochemical sensor platform. By resolving the existing false-positive issue of HRCA, CRISPR Cas12a determines the real positive amplification that able to enhance its sensitivity for extremely low concentrations of nucleic acids and specificity for single-point mutations. In detail, CRISPR Cas12a, which activates the nucleic acid amplification reaction, was used for both trans and cis cleavage for the first time. Finally, selectively amplified DNA was detected using a screen-printed electrode. Using the change in surface coverage by DNA, the electrochemical sensor detected a decrease in the redox signal. In summary, combining a novel DNA amplification method and electrochemical sensor platform, our proposed method compensates for the shortcomings of existing RCA and hyperbranched RCA, secures a high sensitivity of 10 aM, and overcomes false-positivity problems. Moreover, such creative applications of CRISPR Cas12a may lead to the expansion of its applications to other nucleic acid amplification methods.

The development of superior photoelectrochemical (PEC) sensors for biosensing has become a major objective of PEC research. However, conventional PEC-active materials are typically constrained by a weak photocurrent response owing to their limited surface-active sites and high electron-hole recombination rate. Here, a boron and graphene quantum dots codoped g-C3N4 (named GBCN) as PEC sensor for highly sensitive dopamine (DA) detection was fabricated. GBCN exhibited the greatest photocurrent response and PEC activity compared to free g-C3N4 and g-C3N4 doped with boron. The proposed PEC sensor for DA determination exhibited a broad linear range (0.001-800 μM) and a low detection limit (0.96 nM). In particular, a sensitivity up to 10.3771 μA/μM/cm2 was seen in the case of GBCN. The high PEC activity can be attributed to the following factors: (1) the boron and graphene quantum dots co-doping significantly increased the specific surface area of g-C3N4, providing more adsorption sites for DA; (2) the dopants extended the absorption intensity of g-C3N4, red-shifting the absorption from 470 to 540 nm; and (3) the synergism of boron and graphene quantum dots efficiently boosted the photogenerated electrons migration from the conduction band of g-C3N4 to graphene quantum dots, facilitating charge separation. In addition, GBCN also exhibited good anti-interference ability and stability. This research may shed light on the creation of a highly sensitive and selective PEC platform for detecting biomolecules.

The intrinsic biophysical properties of cells, such as mechanical, acoustic, and electrical properties, are valuable indicators of a cell's function and state. However, traditional single-cell biophysical characterization methods are hindered by limited measurable properties, time-consuming procedures, and complex system setups. This study presents acousto-dielectric tweezers that leverage the balance between controllable acoustophoretic and dielectrophoretic forces applied on cells through surface acoustic waves and alternating current electric fields, respectively. Particularly, the balanced acoustophoretic and dielectrophoretic forces can trap cells at equilibrium positions independent of the cell size to differentiate between various cell-intrinsic mechanical, acoustic, and electrical properties. Experimental results show our mechanism has the potential for applications in single-cell analysis, size-insensitive cell separation, and cell phenotyping, which are all primarily based on cells' intrinsic biophysical properties. Our results also show the measured equilibrium position of a cell can inversely determine multiple biophysical properties, including membrane capacitance, cytoplasm conductivity, and acoustic contrast factor. With these features, our acousto-dielectric tweezing mechanism is a valuable addition to the resources available for biophysical property-based biological and medical research.

Imaging a large number of bio-specimens at high speed is essential for many biomedical applications. The common strategy is to place specimens at different lateral positions and image them sequentially. Here we report a new on-chip imaging strategy, termed depth-multiplexed ptychographic microscopy (DPM), for parallel imaging and sensing at high speed. Different from the common strategy, DPM stacks multiple specimens in the axial direction and images the entire z-stack all at once. In our prototype platform, we modify a low-cost car mirror for programmable steering of the incident laser beam. A blood-coated image sensor is then placed underneath the stacked sample for acquiring the resulting diffraction patterns. With the captured images, we perform blind recovery of the incident beam angle and model different layers of the stacked sample as different coded surfaces for object reconstruction. For in vitro experiment, we demonstrate time-lapse cell culture monitoring by imaging 3 stacked microfluidic channels on the coded sensor. For high-throughput cytometric analysis, we image 5 stacked brain sections with a 205-mm2 field of view in ∼50 s. Cytometric analysis is also performed to quantify the cellular proliferation biomarkers on the slides. The DPM approach adds a new degree of freedom for data multiplexing in microscopy, enabling parallel imaging of multiple specimens using a single detector. The demonstrated 6-mm depth of field is among the longest ones in microscopy imaging. The novel depth-multiplexed configuration also complements the miniaturization provided by microfluidics devices, offering a solution for on-chip sensing and imaging with efficient sample handling.

Organ-on-a-chip platforms have potential to offer more cost-effective, ethical, and human-resembling models than animal models for disease study and drug discovery. Particularly, the Blood-Brain-Barrier-on-a-chip (BBB-oC) has emerged as a promising tool to investigate several neurological disorders since it promises to provide a model of the multifunctional tissue working as an important node to control pathogen entry, drug delivery and neuroinflammation. A comprehensive understanding of the multiple physiological functions of the tissue model requires biosensors detecting several tissue-secreted substances in a BBB-oC system. However, current sensor-integrated BBB-oC platforms are only available for tissue membrane integrity characterization based on permeability measurement. Protein secretory pathways are closely associated with the tissue's various diseased conditions. At present, no biosensor-integrated BBB-oC platform exists that permits in situ tissue protein secretion analysis over time, which prohibits researchers from fully understanding the time-evolving pathology of a tissue barrier. Herein, the authors present a platform named "Digital Tissue-BArrier-CytoKine-counting-on-a-chip (DigiTACK)," which integrates digital immunosensors into a tissue chip system and demonstrates on-chip multiplexed, ultrasensitive, longitudinal cytokine secretion profiling of cultured brain endothelial barrier tissues. The integrated digital sensors utilize a novel beadless microwell format to perform an ultrafast "digital fingerprinting" of the analytes while achieving a low limit of detection (LoD) around 100-500 fg/mL for mouse MCP1 (CCL2), IL-6 and KC (CXCL1). The DigiTACK platform is extensively applicable to profile temporal cytokine secretion of other barrier-related organ-on-a-chip systems and can provide new insight into the secretory dynamics of the BBB by sequentially controlled experiments.

Early and accurate diagnosis of acute myocardial infarction (AMI) can significantly reduce patient mortality. A variety of miRNAs are found to dysregulate in AMI patients, but the up- or down-regulation of a specific miRNA may not be evident in the early stage, making it difficult to achieve accurate diagnosis. Here, proposing the design that DNA photonic wire (PW) with no spectral crosstalk would make an excellent template for miRNA conjoint analysis, we report the construction of a miRNA addition probe for the additive analysis of two up-regulated miRNAs (miR-133a and miR-208a) for early diagnosis of AMI in clinical serum samples. A three-dye non-crosstalk DNA PW is built to form the two-step fluorescence resonance energy transfer (FRET) cascade system, in which three paths can blocking the FRET cascade for separate or additive analysis of the two miRNAs. K-Means clustering algorithm is further utilized to classify the output signals of the miRNA addition probe, achieving a 100% accurate diagnosis of early AMI in both the training (n = 40) and validation (n = 19) cohorts of clinical serum samples.

To explore the superiority of multifunctional nanocomposites and realize the joint-detection of foodborne pathogens, an immersible amplification dip-stick immunoassay (DSIA) was exploited for the sensitive detection of Salmonella typhimurium (S. typhi) and Escherichia coli O157:H7 (E. coli O157:H7). Saving for the basic colorimetric performance, the reporter molecule of CoFe2O4 (CFO) possesses multivalent elements (Co2+/3+, Fe2+/3+) as well as multifunction of superior catalase-like activity and magnetic properties. By dint of the catalytic activity of CFO, a directly immersible amplification can be simply achieved to endure the DSIA with an intensive signal and a dual-visible mode for the determination of S. typhi and E. coli O157:H7. In virtue of the magnetic separation and enrichment capability of the CFO, the DSIA can perform a matrix-interference-free detection and obtain a dynamic detection range of 102-108 CFU/mL and a low assay limit of 102 CFU/mL. Moreover, the DSIA has reasonable recovery rates for contamination monitoring of two target bacteria in milk and beef samples. Our research provides a persuasive supplement for the application of multifunctional nanocomposites in the ongoing dip-stick immunoassay and an alternative strategy for the efficient detection of foodborne pathogens.

Practical human biofluid sensing requires a sensor device to differentiate patients from the normal group with high sensitivity and specificity. Label-free molecular identification from human biofluids allows direct classification of abnormal samples, providing insights for disease diagnosis and finding of new biomarkers. Here, we introduce a label-free surface-enhanced Raman scattering sensor based on a three-dimensional plasmonic coral nanoarchitecture (3D-PCN), which has strong electromagnetic field enhancement through multiple hot spots. The 3D-PCN was synthesized on a paper substrate via direct one-step gold reduction, forming a coral-like nanoarchitecture with high absorption property for biofluids. This was fabricated as a urine test strip and then integrated with a handheld Raman system to develop an on-site urine diagnostic platform. The developed platform successfully classified the human prostate and pancreatic cancer urines in a label-free method supported by two types of deep learning networks, with high clinical sensitivity and specificity. Our technology has the potential to be utilized not only for urinary cancer diagnosis but also for various human biofluid sensing systems as a future point-of-care testing platform.

Organs-on-chips (OoCs) are biomimetic in vitro systems based on microfluidic cell cultures that recapitulate the in vivo physicochemical microenvironments and the physiologies and key functional units of specific human organs. These systems are versatile and can be customized to investigate organ-specific physiology, pathology, or pharmacology. They are more physiologically relevant than traditional two-dimensional cultures, can potentially replace the animal models or reduce the use of these models, and represent a unique opportunity for the development of personalized medicine when combined with human induced pluripotent stem cells. Continuous monitoring of important quality parameters of OoCs via a label-free, non-destructive, reliable, high-throughput, and multiplex method is critical for assessing the conditions of these systems and generating relevant analytical data; moreover, elaboration of quality predictive models is required for clinical trials of OoCs. Presently, these analytical data are obtained by manual or automatic sampling and analyzed using single-point, off-chip traditional methods. In this review, we describe recent efforts to integrate biosensing technologies into OoCs for monitoring the physiologies, functions, and physicochemical microenvironments of OoCs. Furthermore, we present potential alternative solutions to current challenges and future directions for the application of artificial intelligence in the development of OoCs and cyber-physical systems. These "smart" OoCs can learn and make autonomous decisions for process optimization, self-regulation, and data analysis.

Charcot-Marie-Tooth disease subtype 1A (CMT1A) is one of the most prevalent demyelinating peripheral neuropathies worldwide, caused by duplication of the peripheral myelin protein 22 (PMP22) gene, which is expressed primarily in Schwann cells (SCs). PMP22 overexpression in SCs leads to intracellular aggregation of the protein, which eventually results in demyelination. Unfortunately, previous biochemical approaches have not resulted in an approved treatment for CMT1A disease, compelling the pursuit for a biophysical approach such as electrical stimulation (ES). However, the effects of ES on CMT1A SCs have remained unexplored. In this study, we established PMP22-overexpressed Schwannoma cells as a CMT1A in vitro model, and investigated the biomolecular changes upon applying ES via a custom-made high-throughput ES platform, screening for the condition that delivers optimal therapeutic effects. While PMP22-overexpressed Schwannoma exhibited intracellular PMP22 aggregation, ES at 20 Hz for 1 h improved this phenomenon, bringing PMP22 distribution closer to healthy condition. ES at this condition also enhanced the expression of the genes encoding myelin basic protein (MBP) and myelin-associated glycoprotein (MAG), which are essential for assembling myelin sheath. Furthermore, ES altered the gene expression for myelination-regulating transcription factors Krox-20, Oct-6, c-Jun and Sox10, inducing pro-myelinating effects in PMP22-overexpressed Schwannoma. While electroceuticals has previously been applied in the peripheral nervous system towards acquired peripheral neuropathies such as pain and nerve injury, this study demonstrates its effectiveness towards ameliorating biomolecular abnormalities in an in vitro model of CMT1A, an inherited peripheral neuropathy. These findings will facilitate the clinical translation of an electroceutical treatment for CMT1A.

Many novel susceptibility tests are being developed to tackle the worldwide problem of antimicrobial resistance (AMR). The key driver behind these developments, that is the need to reduce the response time, requires an understanding of which bacterial characteristic needs to be monitored to provide a rapid and ideally universal signature of susceptibility. Many characteristics have already been studied, most notably bacterial growth, metabolism and motility. Here, we consider electrical impedance to directly access bacterial metabolism, which can be considered a fundamental indicator of bacterial viability. By studying the electrical response of individual bacteria to an antibiotic challenge, we detect antimicrobial action close to its biological limit. Specifically, we find that it takes 30-60 min to register significant changes in impedance for clinical concentrations of antibiotics, in line with other rapid indicators. Our findings suggest that 60 min is the fundamental lower limit of response time for a realistic susceptibility test at clinically relevant antibiotic concentrations.

Gas sensing based on graphene field-effect transistors (GFETs) has gained broad interest due to their high sensitivity. Further progress in gas sensing with GFETs requires to detection of various odor molecules for applications in the environmental monitoring, healthcare, food, and cosmetic industries. To develop the ubiquitous odor-sensing system, establishing an artificial sense of smell with electronic devices by mimicking olfactory receptors will be key. Although the application of olfactory receptors to GFETs is straightforward for odor sensing, synthetic molecules with a similar function to olfactory receptors would be desirable to realize the robust performance of sensing. In this work, we designed three new peptides consisting of two domains: a bio-probe to the target molecules and a molecular scaffold. These peptides were rationally designed based on a motif sequence in olfactory receptors and self-assembled into a molecular thin film on GFETs. Limonene, methyl salicylate, and menthol were employed as representative odor molecules of plant flavors to demonstrate the biosensing of odor molecules. The conductivity change of GFETs against the binding to odor molecules with various concentrations and the dynamic response revealed a distinct signature of three different peptides against individual species of the target molecules. The kinetic response of each peptide exhibited characteristic time constants in the adsorption and desorption process, also supported by the principal component analysis. Our demonstration of the graphene odor sensors with the designed peptides opens a way to establish future peptide-array sensors with multi-sequence of peptide, realizing an odor sensing system with higher selectivity.

Staphylococcus aureus is one of the most prevalent threats to public health. Rapid detection with high sensitivity and targeted killing is crucial to curb its spread. Herein, a metal-bearing nanocomposite, consisting of a bimetallic nanoparticle and a metal-organic framework (Au/Ir@Cu/Zn-MOF) was constructed. Upon conjugation with anti-S. aureus antibody, this nanocomposite (Ab-Au/Ir@Cu/Zn-MOF) was exploited for its dual functions, i.e. as a reporting probe in a lateral flow immunoassay and a high efficiency antibacterial reagent. Benefiting from the enrichment of Au/Ir NPs by the Cu/Zn-MOF, the Au/Ir@Cu/Zn-MOF-based lateral flow immunoassay sensor exhibited a visual limit of detection of 103 CFU/mL, which was100 times more sensitive than Au/Ir-based sensor. Moreover, the Ab-Au/Ir@Cu/Zn-MOF probe possessed synergistic photothermal-chemodynamic bactericidal effect that specifically targeted against S. aureus. Under a co-treatment by H2O2 (0.4 mM) and 808 nm near infrared irradiation (1 W/cm2, 5 min), complete sterilization of 5 × 105-106 CFU/mL S. aureus was achieved at a nanocomposite concentration as low as 6.25 μg/mL. The superior antibacterial efficiency was attributable to the three-fold properties of the Ab-Au/Ir@Cu/Zn-MOF probe: (1) enhanced multi-enzyme mimicking activities that promote reactive oxygen species generation, (2) high photothermal activity (efficiency of 53.70%), and (3) bacteria targeting ability via the antibody coating. By changing the antibody, this nanocomposite can be tailored to target a wide range of bacteria species, for detection and for precise antibacterial treatment.

Methyl salicylate (MeSal) is an organic compound present in plants during stress events and is therefore a key marker for early plant disease detection. It has usually been detected by conventional methods that require bulky and costly equipment, such as gas chromatography or mass spectrometry. Currently, however, chemical sensors provide an alternative for MeSal monitoring, showing good performance for its determination in the vapour or liquid phase. The most promising concepts used in MeSal determination include sensors based on electrochemical and conductometric principles, although other technologies based on mass-sensitive, microwave, or spectrophotometric principles also show promise. The receptor elements or sensitive materials are shown to be part of the key elements in these sensing technologies. A literature survey identified a significant contribution of bioreceptors, including enzymes, odourant-binding proteins or peptides, as well as receptors based on polymers or inorganic materials in MeSal determination. This work reviews these concepts and materials and discusses their future prospects and limitations for application in plant health monitoring.

Electrochemiluminescence (ECL) has several advantages, such as a near-zero background signal, high sensitivity, wide dynamic range, simplicity, and is widely used for sensing, imaging, and single cell analysis. ECL luminophores are the key factors in the performance of various applications. Among various luminophores, small organic luminophores exhibit many intriguing features including good biocompatibility, facile modification, well-defined molecular structure, and sustainable raw materials, making small organic luminophores attractive for the use in the ECL field. Although many great achievements have been made in the synthesis of new small organic luminophores, solving various challenges, and expanding new applications, there are almost no comprehensive reviews on small organic ECL luminophores. In this review, we briefly introduce the advantages and emission mechanisms of small organic ECL luminophores, summarize the main types, molecular characteristics, and ECL properties of most existing small organic ECL luminophores, and present the important applications and design principles in sensors, imaging, single cell analysis, sterilization, and other fields. Finally, the challenges and outlook of organic ECL luminophores to be popularized in biosensing applications are also discussed.

Herein, we introduce wearable potentiometric biosensors on screen-printed carbon electrodes (SPCEs) for on-body and on-site monitoring of urea in sweat. The biosensor architecture was judiciously designed to detect urea at different pHs and incorporate a pH sensor, thus containing polyaniline ink, urease bioink and a polyvinylchloride membrane. Urea detection could be performed in the wide range from 5 to 200 mM at pH 7.0, encompassing urea levels in human sweat. The biosensor response was fast (incubation time 5 min), with no interference from other substances in sweat. Reliable urea detection could be done in undiluted human sweat with a skin-worn flexible device using the pH correction strategy afforded by the pH sensor. The performance of the epidermal biosensor was not affected by severe bending strains. The feasibility of mass production was demonstrated by fabricating epidermal flexible biosensors using slot-die coating with a roll-to-roll technique.

As a key factor in cellular metabolic processes, nitric oxide (NO) is a challenging target for in situ real-time monitoring due to its transient property and short diffusion distance. Scanning electrochemical microscopy (SECM) has unique advantages in single-cell analysis, which can obtain the electrochemical current by scanning the cell surface with a tip microelectrode. In particular, it can further improved the electrochemical response by enhancing the interface properties of its tip. Here, an interface design strategy based on platinum single nanoparticle (Pt NP) was developed, and fluorinated self-assembled monolayers (SAMs) were used to further improve its performance. This modified tip was used as an SECM probe for NO concentration monitoring and morphological imaging of single MCF-7 cells. It has the high sensitivity (164.7 μA/μM·cm2) and good selectivity for NO detection, which benefits from the efficient catalytic properties of Pt NPs and high mass transport and hydrophobic antifouling properties of the interface. Notably, it shows a superior performance in detecting the fluctuation of NO released by a single MCF-7-cell under the stimulation of cadmium (Cd), which demonstrates a promising method for using a single-particle-based tip in SECM applications.

Prostate specific membrane antigen (PSMA)-positive exosomes have the potential to serve as highly sensitive biomarkers for prostate cancer detection. Herein, a sensitive electrochemical biosensor for the ultrasensitive detection of PSMA-positive exosomes has been constructed based on a peptide-templated AgNPs nanoprobe. In this work, PSMA-specific binding peptides immobilized on a gold electrode were responsible for prostate cancer-derived exosomes capturing. Well-designed peptide (CCY- LWYIKC) serves a dual role: as a signal probe and as a recognizer in the exosomes-identification process. Specifically, LWYIKC bind to cholesterol at the exosome membranes, and CCY function as peptide templates to host a large number of silver nanoparticles, leading to a strong electrochemical signal. Thus, the concentration of exosomes can be quantified via electrochemical signal. This innovative method displayed a wide detection range of 102 to 108 particles/μL and a detection limit as low as 37 particles/μL. Notably, the method has shown outstanding performance when validated using clinical samples, suggesting its potential for clinical applications.

The conventional heating, ventilation, and air conditioning (HVAC) systems are based on a set-point control approach that only considers the temperature of the environment without reflecting the thermophysiological status of the occupant. This approach not only fails to fully satisfy individual thermal preferences, but it also makes an HVAC operation energy-inefficient. One possible solution is to control the indoor thermal condition based on an accurate prediction of the occupant's thermal comfort to prevent any unnecessary energy consumption. Here, we present an artificial intelligence (AI) wearable sensor-based human-in-the-loop HVAC control system that is operated on a real-time basis reflecting the thermophysiological condition of the occupant to automatically improve their thermal comfort while reducing the energy consumption of the building. The wristband-type, AI-based, three-point wearable temperature sensor offers excellent thermal comfort prediction accuracy (93.9%), enabling a human-centric HVAC control operation. A proof-of-concept demonstration of closed human-in-the-loop HVAC control using the AI-enabled wearable sensor system confirms both the accuracy of the thermal comfort prediction and the energy-efficiency of this approach, demonstrating its potential as a new solution that improves the occupant's thermal comfort and provides building energy savings.

Fluorescent nanosensor-based tumor imaging holds great promise in cancer diagnosis and treatment assistance, yet the signal contrast is heavily hampered by the unspecific/unwanted activation at microscopic regions with a highly restricted local abundance of biomarkers. Herein, we developed an activation boosting strategy by the integration and manipulation of dual-factor coactivation of sensing and lysosome escape facilitated the rise of cytosolic biomarker accessibility. By employing hybrid DNA probes on gold nanoquenchers, ATP sensing initiated conformation switch of the corresponding aptamer units triggered the exposure of a hidden toehold in a loop structure. Sequentially, miRNA-21 sensing was triggered by toehold-mediated strand displacement and detachment of the binding complexes. The application of lysosomotropic agent chloroquine at optimized time interval facilitated the release of nanosensors into the cytosol and a ∼10.5-fold increment of intracellular fluorescence in vitro, while coactivation improved the cancer-to-normal cell signal ratio by ∼5.9 times. The synergy effects led to a high tumor-to-normal tissue ratio value of ∼7.9 in the in vivo imaging results. This strategy establishes a new paradigm of fluorescent nanosensors for selective and specific tumor imaging.

Considering the fact that acute myocardial infarction has shown a trend towards younger age and has become a major health problem, it is necessary to develop rapid screening devices to meet the needs of community health care. Herein, we developed an artificial neural network-assisted solar-powered photoelectrochemical (SP-PEC) sensing platform for rapid screening of cardiac troponin I (cTnI) protein in the prognosis of patients with acute myocardial infarction (AMI) by integrating a self-powered photoelectric signal output system with low-cost screen-printed paper electrodes functionalized with ultrathin Bi2O2S (BOS) nanosheets. An integrated solar-powered PEC immunoassay with micro-electro-mechanical system (MEMS) was constructed without an excitation light source. The quantification of cTnI protein was obtained by the electrical signal changes caused by the electro-oxidation process of H2O2, generated by the classical split immune reaction, on the electrode surface. The test electrodes were developed as dual working electrodes, one for target cTnI testing and the other for evaluating light intensity, to reduce the temporal inconsistency of sunlight. The photoelectrodes were discovered to exhibit satisfactory negative response to target concentrations in the dynamic range of 2.0 pg mL-1-10 ng mL-1 since being regressed in an improved artificial neural network (ANN) model using the pooled dataset of target signals affected by the light source. The difference of hot electron and hole transfer behavior in different thickness of nano-materials was determined by finite element analysis (FEA), which provided a theoretical basis for the development of efficient PEC sensors. This work presents a unique perspective for the design of a revolutionary low-cost bioassay platform by inventively illuminating the PEC biosensor's component process without the use of light.

The current coronavirus disease 2019 (COVID-19) pandemic is caused by several variants of severe acute respiratory syndrome coronavirus-2 virus (SARS-CoV-2). With the roll-out of vaccines and development of new therapeutics that may be targeted to distinct viral molecules, there is a need to screen populations for viral antigen-specific SARS-CoV-2 antibodies. Here, we report a rapid, multiplexed, electrochemical (EC) device with on-chip control that enables detection of SARS-CoV-2 antibodies in less than 10 min using 1.5 μL of a patient sample. The EC biosensor demonstrated 100% sensitivity and specificity, and an area under the receiver operating characteristic curve of 1, when evaluated using 93 clinical samples, including plasma and dried blood spot samples from 54 SARS-CoV-2 positive and 39 negative patients. This EC biosensor platform enables simple, cost-effective, sensitive, and rapid detection of anti-SARS-CoV-2 antibodies in complex clinical samples, which is convenient for evaluating humoral-responses to vaccination or infection in population-wide testing, including applications in point-of-care settings. We also demonstrate the feasibility of using dried blood spot samples that can be collected locally and transported to distant clinical laboratories at ambient temperature for detection of anti-SARS-CoV-2 antibodies which may be utilized for serological surveillance and demonstrate the utility of remote sampling.

Cardiovascular Disease (CVD) is amongst the leading cause of death globally, which calls for rapid detection and treatment. Biosensing devices are used for the diagnosis of cardiovascular disease at the point-of-care (POC), with lateral flow assays (LFAs) being particularly useful. However, due to their low sensitivity, most LFAs have been shown to have difficulties detecting low analytic concentrations. Breakthroughs in artificial intelligence (AI) and image processing reduced this detection constraint and improved disease diagnosis. This paper presents a novel patches-selection approach for generating LFA images from the test line and control line of LFA images, analyzing the image features, and utilizing them to reliably predict and classify LFA images by deploying classification algorithms, specifically Convolutional Neural Networks (CNNs). The generated images were supplied as input data to the CNN model, a strong model for extracting crucial information from images, to classify the target images and provide risk stratification levels to medical professionals. With this approach, the classification model produced about 98% accuracy, and as per the literature review, this approach has not been investigated previously. These promising results show the proposed method may be useful for identifying a wide variety of diseases and conditions, including cardiovascular problems.

Accurate monitoring of cardiomyocyte action potentials (APs) is essential to understand disease propagation and for trials of novel therapeutics. Patch clamp techniques offer 'gold standard' measurements in this field, but are notoriously difficult to operate and only provide measurements of a single cell. Here we propose photoelectrochemical imaging (PEI) with light-addressable potentiometric sensors (LAPS) in conjunction with a setup for controlling the contact force between the cardiomyocyte organoids and the sensor surface for measuring APs with high sensitivity. The method was validated through measuring the responses to drugs, and the results successfully visualized the expected electrophysiological changes to the APs. PEI allows for several cells to be monitored simultaneously, opening further research to the electrophysiological interactions of adjoining cells. This method expands the applications of PEI to three-dimensional geometries and provides the fields of stem cell research, drug trials and heart disease modelling with an invaluable tool to further investigate the role of APs.

In this work, a self-powered sensor was proposed for simultaneous detection of two typical steroid hormones, namely cortisol (COR) and progesterone (P4). A tri-channel photofuel cell (PFC) consisting of three spatially resolved SnS2@SnO2 photoanodes and one Pt cathode was designed to generate the electricity to drive the sensing process under the control of a multiplex switch. Among three photoanodes, one served as the control, while the other two were modified with COR-binding or P4-binding aptamer to respond specifically to the COR or P4 target. The ratios of the inhibited PFC output from aptamer-immobilized photoanodes to the reference signal from the control photoanode were utilized for simultaneous detection of COR and P4. The results showed that the developed self-powered sensor exhibited broad concentration ranges toward targets, with COR concentration ranging from 1 nM to 1000 nM and P4 concentration ranging from 1 nM to 500 nM. The detection limits for COR and P4 were calculated to be 0.88 nM and 0.52 nM, respectively. Moreover, the proposed sensing platform demonstrated high selectivity, good reproducibility, and high stability. Finally, the sensor was successfully applied to the simultaneous determination of COR and P4 in a human female serum sample.

Quantifying biomarkers at the early stage of the disease is challenging due to the low abundance of biomarkers in the sample and the lack of sensitive techniques. This article reports the development of a novel microfluidic electrochemical biosensing platform to address this challenge. The electrochemical sensing is achieved by utilizing a micropillar array electrode (μAE) coated with 3D bimetallic Pt-Pd nanotrees to enhance the sensitivity. A bubble-based acoustic microstreaming technique is integrated with the device to increase the contact of analyte molecules with the surface of electrodes to further enhance the electrochemical performance. The current density of Pt-Pd NTs/μAE with acoustic microstreaming is nearly 22 times that of the bare planar electrode in potassium ferrocyanide solution. The developed biosensor has demonstrated excellent sensing performance. For hydrogen peroxide detection, both the Pt-Pd NTs/μAE and acoustic microstreaming contribute to the sensitivity enhancement. The current density of the Pt-Pd NTs/μAE is approximatively 28 times that of the bare μAE. With acoustic microstreaming, this enhancement is further increased by nearly 1.6 times. The platform has a linear detection range of 5-1000 μM with a LOD of 1.8 μM toward hydrogen peroxide detection, while for sarcosine detection, the linear range is between 5 and 100 μM and LOD is 2.2 μM, respectively. Furthermore, the sarcosine biosensing shows a high sensitivity of 667 μA mM-1∙cm-2. Such a sensing platform has the potential as a portable device for high sensitivity detection of biomarkers.

Conventional liquid crystal (LC)-based biosensors utilize predominantly thermotropic LCs as the signal-transducing media, which are less environmentally sustainable compared with lyotropic counterparts. In this study, the nematic phase of the anionic azo dye sunset yellow (SSY), a type of lyotropic chromonic liquid crystals (LCLCs), was employed in the optical and electrical biosensing of bovine serum albumin (BSA) and the cancer biomarker CA125. The optical response observed under a polarizing optical microscope was quantified by image analysis, taking advantage of the specific absorption of SSY. The electrical response derived from the dielectric spectra of SSY provided a new alternative for quantitative bioassay based on nematic LCLCs. The limit of detection (LOD) of the optical and electrical protein assay was ∼10-11- and ∼10-10-g/ml BSA, respectively, whereas that of the optical and electrical immunoassay was 5.97 × 10-11 and 6.02 × 10-12 g/ml for CA125, respectively. Moreover, real-time monitoring and kinetic analysis, which are hardly achievable for the hydrophobic thermotropic LCs, were demonstrated by dispersing CA125 in nematic SSY and subsequently recording the optical response over time during the specific binding between CA125 and the immobilized anti-CA125 antibody. Results from this study further the potential of nematic LCLCs in biosensing, especially in dielectric and real-time detection.

The current organ-on-chip platforms used for studying respiratory drug delivery are limited to the administration of drug solutions and suspensions, lacking the in vivo aerosol drug administration and aerosol interaction with the respiratory tract barrier. Moreover, they mostly rely on conventional assays that require sample collection and 'off the chip' analyses, which can be labor-intensive and costly. In this study, a human nasal epithelial mucosa (NEM)-on-a-chip is developed that enables the deposition of aerosolized nasal formulations while emulating realistic shear stresses (0.23 and 0.78 Pa), exerted to the inferior and middle turbinate of the human nasal cavity. Under these different dynamic conditions in the donor channel of the NEM-on-a-chip, the deposited dose of aerosols and particle size distributions varied. In addition, the increase in the shear stress to 0.78 Pa adversely affected the cells' viability, reflected by a 36.9 ± 5.4% reduction in the transepithelial electrical resistance. The epithelial transport profiles of aerosolized ibuprofen formulations under 0.23 Pa shear stress were successfully monitored in real-time by an electrochemical sensor embedded in the acceptor channel, where the NEM-on-a-chip was able to monitor the effect of permeation enhancer in the test formulation on the rate of drug transport. The novel NEM-on-a-chip can potentially be a promising physiologically relevant tool for reliable nasal aerosol testing in vitro.

To differentiate whole wheat foods from refined wheat foods is still challenging grain industry and confusing consumers. Alkylresorcinols (ARs), as biomarkers of whole wheat grains, can serve for assessing the authenticity of whole wheat foods. Herein, a highly efficient fluorescence sensing platform (CDs@MIP) for rapid and sensitive analysis of ARs was explored, using carbon dots (CDs) as fluorophores and 5-heneicosylresorcinol (C21:0 AR) as template molecules embedded in a molecularly imprinted polymer (MIP) coating. Benefiting from the specific cavities in the probe and a photo-induced electron transfer effect, the fluorescence intensity of CDs@MIP was significantly quenched in the presence of C21:0 AR, exhibiting a superior binding efficiency and selectivity. As a result, the fabricated optical sensor delivered a wide linear range of C21:0 AR from 0.015 to 60 μg mL-1 with an ultralow detection limit of 4 ng mL-1. It was noteworthy that the sensor was successfully applied for the rapid detection of C21:0 AR in commercial whole-wheat foods as well as visualization analysis on the test paper, comprehensively validating the practicality and efficacy of CDs@MIP based fluorescence assay. The study provides a rapid and sensitive detection method of C21:0 AR, paving a new way for guiding grain industry to effectively qualify the authenticity and to quantify the content of whole wheat in wheat-based foods.

Indium tin oxide, semiconductor nanomaterial ZnO, and Cu2O were first loaded on the surface of the optical fiber to form an optical fiber probe. Large-volume macroscopic spatial light is replaced by an optical fiber path, and remote light injection is implemented. Based on the optical fiber probe, a photoelectrochemical biosensor was constructed and remote detection of cysteine was realized. In this tiny device, the optical fiber probe not only acts as a working electrode to react with the analyte but also directs the light exactly where it is needed. Simultaneously, the electrochemical behavior of cysteine on the surface of the working electrode is dominated by diffusion-control, which provides strong support for quantitative detection. Then, under the bias potential of 0 V, the linear range of the fiber-optic-based cysteine biosensor was 0.01∼1 μM, the regression coefficient (R2) value was 0.9943. In spiked synthetic urine, the detection of cysteine was also realized by the integrated biosensor. Moreover, benefiting from the low optical fiber loss, the new structure also possesses a unique remote detection function. This work confirms that photoelectrochemical biosensors can be integrated via optical fibers and retain comparable sensing performance. Based on this property, different materials can also be loaded on the surface of the optical fiber for remote detection of other analytes. It is expected to facilitate the research on fiber-optic-based integrated biosensors and show application prospects in diverse fields such as biochemical analysis and disease diagnosis.

Efficient capture of circulating tumor cells (CTCs) from cancer patients is an important technique that may promote early diagnosis and prognosis monitoring of cancer. However, the existing systems have certain disadvantages, such as poor selectivity, low capture efficiency, consumption of antibodies, and difficulty in release of CTCs for downstream analysis. Herein, we fabricated an innovative PEGylated boronate affinity cell imprinted polydimethylsiloxane (PBACIP) for highly efficient capture of CTCs from cancer patients. The antibody-free PBACIP possessed hierarchical structure of imprinted cavities, which were inlaid with boronic acid modified SiO2 nanoparticles (SiO2@BA), so it could specifically capture target CTCs from biological samples due to the synergistic effect of boronate affinity and cell imprinting. Furthermore, PEGylation was accurately completed in the non-imprinted region by the template cells occupying the imprinted cavity, which not only retained the microstructure of original imprinted cavities, but also endowed PBACIP with hydrophilicity. The artificial PBACIP could efficiently capture human breast-cancer cells from biological sample. When 5 to 500 SKBR3 cells were spiked in 1 mL mice lysed blood, the capture efficiency reached 86.7 ± 11.5% to 96.2 ± 2.3%. Most importantly, the PBACIP was successfully used to capture CTCs from blood of breast cancer patients, and the captured CTCs were released for subsequent gene mutation analysis. The PBACIP can efficiently capture and release CTCs for downstream analysis, which provides a universal strategy toward individualized anti-tumor comprehensive treatments and has great potential in the future cell-based clinical applications.

The frequent emergence of SARS-CoV-2 variants increased viral transmissibility and reduced protection afforded by vaccines. The rapid, multichannel, and intelligent screening of variants is critical to minimizing community transmissions. DNA molecular logic gates have attracted wide attention in recent years due to the powerful information processing capabilities and molecular data biocomputing functions. In this work, some molecular switches (MSs) were connected with each other to implement arbitrary binary functions by emulating the threshold switching of MOS transistors and the decision tree model. Using specific sequences of different SARS-CoV-2 variants as inputs, the MSs net was used to build several molecular biocomputing circuits, including NOT, AND, OR, INHIBIT, XOR, half adder, half subtractor, full adder, and full subtractor. Four fluorophores (FAM, Cy3, ROX, and Cy5) were employed in the logic systems to realize the multichannel monitoring of the logic operation results. The logic response is fast and can be finished with 10 min, which facilitates the rapid wide-population screening for SARS-CoV-2 variants. Importantly, the logic results can be directly observed by the naked eye under a portable UV lamp, thus providing a simple and intelligent method to enable high-frequency point-of-care diagnostics, particularly in low-resource communities.

Point-of-care testing (POCT) of blood cell count (BCC) is an emerging approach that allows laypersons to identify and count whole blood cells through simple manipulation. To date, POCTs for BCC were mainly achieved by "stationary" images through blood smears or single-laity arranged cells in the microwell, making it difficult to obtain statistically sufficient numbers of cells. In this work, we present a fully integrated POCT device solely using "in-flow" imaging of 3 μL fingertip whole blood for improved identification and counting accuracy of BCC analysis. A miniaturized magnetic stirring module was integrated to maintain the temporal stability of cell concentration. A relatively high throughput (∼8000 cells/min) with a 30-fold dilution ratio of whole blood can be tested for as long as 1 h to examine sufficient numbers of cells, and the subclass cell concentration keeps constant. To improve the identification accuracy, multi-frame "in-flow" imaging was used to track the cell motion trails with multi-angle morphology analysis. This proof-of-concept was then validated with healthy whole blood samples and 75 cases of clinical patients with abnormal concentrations of red blood cells (RBCs), white blood cells (WBCs), and platelets (PLT). The average precision (AP) value of WBCs identification was improved from 0.8622 to 0.9934 using the multi-frame analysis method. And the high fitting degrees (>0.98) between our POCT device and the commercial clinical equipment indicated good agreement. This POCT device is user-friendly and cost-effective, making it a potential tool for diagnosing abnormal blood cell morphology or concentration in the field setting.

The accurate determination of tumor biomarkers in blood is of vital significance in the diagnosis and therapy of tumor disease. In this research, an innovative sandwich-type electrochemical immunosensor is designed for the ultrasensitive determination of tumor biomarker AFP using spherical nucleic acids-templated silver nanoclusters (AgNCs) sensing platform. For this purpose, on one hand, DNA functionalized gold nanoparticles (AuNPs@DNA) is selected not only as the cross-linker to immobilize the primary antibody (anti-AFP antibody 1, Ab1) to obtain AuNPs@DNA-Ab1, but also as the template for synthesizing AgNCs on AuNPs to form AuNPs@DNA-AgNCs. On the other hand, p-sulfonated calix[4]arene (pSC4) modified Au is chosen to immobilize the secondary antibody (anti-AFP antibody 2, Ab2) through host-guest recognition between Ab2 and pSC4. When AFP is encountered, the immunoreaction signal can be significantly amplified by the electrochemical reduction of AgNCs. Under optimal circumstances, the sandwich-type electrochemical immunosensor exhibits broad limit of linearity from 0.001 to 100 ng mL-1 (R2 = 0.997) and low detection limit of 7.74 fg mL-1 (S/N = 3). The immunosensor possesses excellent repeatability and selectivity, offering a novel method for sensitive clinical diagnosis of tumor markers in human hepatocellular carcinoma.

The number of synthetic biology-based solutions employed in the medical industry is growing every year. The whole cell biosensors being one of them, have been proven valuable tools for developing low-cost, portable, personalized medicine alternatives to conventional techniques. Based on this concept, we targeted one of the major health problems in the world, Chronic Kidney Disease (CKD). To do so, we developed two novel biosensors for the detection of two important renal biomarkers: urea and uric acid. Using advanced gene expression control strategies, we improved the operational range and the response profiles of each biosensor to meet clinical specifications. We further engineered these systems to enable multiplexed detection as well as an AND-logic gate operating system. Finally, we tested the applicability of these systems and optimized their working dynamics inside complex medium human blood serum. This study could help the efforts to transition from labor-intensive and expensive laboratory techniques to widely available, portable, low-cost diagnostic options.

The development of novel biomedical sensors as highly promising devices/tools in early diagnosis and therapy monitoring of many diseases and disorders has recently witnessed unprecedented growth; more and faster than ever. Nonetheless, on the eve of Industry 5.0 and by learning from defects of current sensors in smart diagnostics of pandemics, there is still a long way to go to achieve the ideal biomedical sensors capable of meeting the growing needs and expectations for smart biomedical/diagnostic sensing through eHealth systems. Herein, an overview is provided to highlight the importance and necessity of an inevitable transition in the era of digital health/Healthcare 4.0 towards smart biomedical/diagnostic sensing and how to approach it via new digital technologies including Internet of Things (IoT), artificial intelligence, IoT gateways (smartphones, readers), etc. This review will bring together the different types of smartphone/reader-based biomedical sensors, which have been employing for a wide variety of optical/electrical/electrochemical biosensing applications and paving the way for future eHealth diagnostic devices by moving towards smart biomedical sensing. Here, alongside highlighting the characteristics/criteria that should be met by the developed sensors towards smart biomedical sensing, the challenging issues ahead are delineated along with a comprehensive outlook on this extremely necessary field.

Carbon monoxide (CO) is not only a highly poisonous gas that brings great health risk, but also a significant signaling molecule in body. However, it is still challengeable for development of alternative colorimetric probes to traditional organic chromophores for simple, sensitive and convenient CO sensing. Here, for the first time, we rationally design a novel hydrophilic AgPt-Fe3O4 nanozyme with a unique heterodimeric nanostructure for colorimetric sensing of CO based on the excellent peroxidase-like catalytic activity as well as highly poisonous effect of CO on the nanozyme's catalytic activity. Both experimental evidence and theoretical calculations reveal the trimetallic AgPt-Fe3O4 nanozyme is susceptible to poisoning with the strongest affinity towards CO compared to individual Fe3O4 or Ag-Fe3O4, which is attributed to the adequate exposure of the active metallic sites and efficient interfacial synergy of unique heterodimeric nanostructure. Accordingly, a novel nanozyme-based colorimetric strategy is developed for CO detection with a low detection limit of 5.6 ppb in solution. Furthermore, the probe can be prepared as very convenient test strips and integrated with the portable smartphone platforms for detecting CO gas samples with a low detection limit of 8.9 ppm. Overall, our work proposes guidelines for the rational design of metallic heterogeneous nanostructure to expand the analytical application of nanozyme.

Recent advances in enzymatic biofuel cells (EBFCs) have resulted in great progress in health monitoring and supplying power to medical applications, such as drug delivery. On the other hand, to enhance the electric field-assisted transdermal permeation for facial mask application, an external power source is usually required. Herein, we attempted to combine an EBFC with a facial mask so that the microcurrent generated can boost the transdermal permeability of target molecules in the facial mask essence. When screen-printed onto a polypropylene-based non-woven fabric, the three-layered flexible EBFC could produce a voltage of ∼0.4 V and a maximum power density of 23.3 μW cm-2, leading to an approximately 2-3-fold increase in permeated nicotinamide, arbutin, and aspirin levels within 15 min compared to non-iontophoretic transdermal drug delivery. Both cell viability and animal experiments further demonstrated that the EBFC-powered iontophoresis worked well in living animals with good biocompatibility. These results suggest that the EBFC-powered iontophoretic facial mask can effectively improve the permeation of drugs and holds a promise for the possible cosmetic application.

As one of the most toxic chemical substances, aflatoxin B1 (AFB1) has a strong carcinogenic effect even at a trace level in human and animal, which severely threatens human health and even causes cancers. Therefore, ultrasensitive detection of AFB1 is of significant importance. For such analysis, dual II-scheme sheet-like Bi2S3/Bi2O3/Ag2S heterostructures were prepared by the in-situ growth method, which exhibited high separation efficiency for the electron-hole (e--h+) pairs, prominent stability, and high photoactivity. Moreover, the dendritic nanorod-like Au@Pd@Pt (Au@Pd@Pt DNRs) nanozyme was homely synthesized, whose peroxidase-like activity was scrupulously investigated by catalytical oxidation of diaminobenzidine (DAB) in the presence of H2O2. Integration by the aptasensing strategy, a photoelectrochemical (PEC) "signal-on" aptasensor was prepared, which exhibited a broader linear range of 0.5 pg mL-1-100 ng mL-1 with a lower limit of detection (LOD = 0.09 pg mL-1, S/N = 3). This work provides a feasible strategy to develop advanced PEC biosensors for actual analysis of environmental pollutants.

Skin-centric diagnosis techniques, such as epidermal physiological parameter monitoring, have developed rapidly in recent years. The analysis of interstitial fluid (ISF), a body liquid with abundant physiological information, is a promising method to obtain health status because ISF is easily assessed by implanted or percutaneous measurements. Reverse iontophoresis extracts ISF by applying an electric field onto the skin, and it is a promising method to noninvasively obtain ISF, which, in turn, enables noninvasive epidermal physiological parameter monitoring. However, the development of reverse iontophoresis was relatively slow around the 2010s due to the rigidity and low biocompatibility of the applied devices. With the rapid development of flexible electronic technology in recent years, new progress has been made in the field of reverse iontophoresis, especially in the field of blood glucose monitoring and drug monitoring. This review summarizes the recent advances and discusses the challenges and opportunities of reverse iontophoresis.

Effective isolation and in-depth analysis of Circulating Tumour Cells (CTCs) are greatly needed in diagnosis, prognosis and monitoring of the therapeutic response of cancer patients but have not been completely fulfilled by conventional approaches. The rarity of CTCs and the lack of reliable biomarkers to distinguish them from peripheral blood cells have remained outstanding challenges for their clinical implementation. Herein, we developed a high throughput Static Droplet Microfluidic (SDM) device with 38,400 chambers, capable of isolating and classifying the number of metabolically active CTCs in peripheral blood at single-cell resolution. Owing to the miniaturisation and compartmentalisation capability of our device, we first demonstrated the ability to precisely measure the lactate production of different types of cancer cells inside 125 pL droplets at single-cell resolution. Furthermore, we compared the metabolomic activity of leukocytes from healthy donors to cancer cells and showed the ability to differentiate them. To further prove the clinical relevance, we spiked cancer cell lines in human healthy blood and showed the possibility to detect the cancer cells from leukocytes. Lastly, we tested the workflow on 8 preclinical mammary mouse models including syngeneic 67NR (non-metastatic) and 4T1.2 (metastatic) models with Triple-Negative Breast Cancer (TNBC) as well as transgenic mouses (12-week-old MMTV-PyMT). The results have shown the ability to precisely distinguish metabolically active CTCs from the blood using the proposed SDM device. The workflow is simple and robust which can eliminate the need for specialised equipment and expertise required for single-cell analysis of CTCs and facilitate on-site metabolic screening of cancer cells.

In order to improve the recognition performance of MIPs sensors in chiral drug enantiomers, a novel a highly selective molecular recognition method based on protein-assisted immobilization of chiral molecular conformation was developed. S-fluoxetine (S-FLX) as the target chiral molecule, human serum albumin (HSA), which has a high affinity and strong interactions with S-FLX, was screened from 11 proteins to serve as an auxiliary recognition unit for the fixation of chiral conformation. By incorporating HSA into the preparation of molecularly imprinted polymers (MIPs), the natural chirality and high stereoselectivity of the protein were leveraged for the induction and fixation of the stereo conformation of S-FLX, refinement of internal structures of the imprinted cavities. The sensor exhibited excellent chiral recognition ability and high detection sensitivity. The changes of probe signal intensity of the MIPs/HSA sensor were positively correlated with the logarithmic concentration of S-FLX in the range of 1.0 × 10-16-1.0 × 10-11 mol L-1, where a detection limit of 6.43 × 10-17 mol L-1 was achieved (DL = 3δb/K). The selectivity of MIPs/HSA sensor in recognizing S-FLX was increased by 18.5 times and the sensitivity was increased by 2.6 times after the incorporation of HSA. The developed sensor was successfully used for the analysis of S-FLX in fluoxetine hydrochloride capsules.

The ectopic co-expression of taste and olfactory receptors in cardiomyocytes provides not only possibilities for the construction of biomimetic gustatory and olfactory sensors but also promising novel therapeutic targets for tachycardia treatment. Here, bitter taste and olfactory receptors endogenously expressed in HL-1 cells were verified by RT-PCR and immunofluorescence staining. Then HL-1 cardiomyocyte-based integrated gustatory and olfactory sensing array coupling with the microelectrode array (MEA) was first constructed for drugs screening and evaluation for tachycardia treatment. The MEA sensor detected the extracellular field potentials and reflected the systolic-diastolic properties of cardiomyocytes in real time in a label-free and non-invasive way. The in vitro tachycardia model was constructed using isoproterenol as the stimulator. The proposed sensing array facilitated potential drug screening for tachycardia treatment, such as salicin, artemisinin, xanthotoxin, and azelaic acid which all activated specific receptors on HL-1 cells. IC50 values for four potential drugs were calculated to be 0.0036 μM, 309.8 μM, 14.68 μM, and 0.102 μM, respectively. Visualization analysis with heatmaps and PCA cluster showed that different taste and odorous drugs could be easily distinguished. The mean inter-class Euclidean distance between different bitter drugs was 1.681, which was smaller than the distance between bitter and odorous drugs of 2.764. And the inter-class distance was significantly higher than the mean intra-class Euclidean distance of 1.172. In summary, this study not only indicates a new path for constructing novel integrated gustatory and olfactory sensors but also provides a powerful tool for the quantitative evaluation of potential drugs for tachycardia treatment.

A novel metal-organic framework (MOF)/covalent-organic framework (COF) heterostructure hybrid composite (NH2-UiO-66/TpPa-1-COF) with excellent photoactivity was developed, which further acted as the photoelectrochemical sensitized layer of a molecularly imprinted photoelectrochemical (MIP-PEC) sensor for extremely sensitive and selective determination of dibutyl phthalate (DBP). The NH2-UiO-66/TpPa-1-COF was synthesized using a simple one-step solvothermal method, which showed improved photocurrent response owing to heterojunction formation, favorable energy-band configuration and strong light absorption capacity. To improve the sensing performance, molecularly imprinted polymer (MIP) was developed by sol-gel polymerization method as the recognition component of PEC sensor. The specific binding of imprinting sites towards DBP could block the electron transfer, causing decreased photocurrent response of the MIP-PEC sensor. The MIP-PEC sensor showed a wide detection range from 0.1 nmol L-1 to 100 μmol L-1 with a limit of detection of 3.0 × 10-11 mol L-1 under optimal conditions. Meanwhile, the proposed MIP-PEC sensor showed good stability, selectivity, reproducibility, and applicability in real samples. This is the first attempt to apply MOF/COF heterostructure hybrid composite for MIP-PEC sensor construction, providing new insight into the potential applications of microporous crystalline framework heterostructure hybrid composite in the sensing field.