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The development of robust biosensing strategies that can be easily implemented in everyday life remains a challenge for the future of modern biosensor research. While several reagentless approaches have attempted to address this challenge, they often achieve user-friendliness through sacrificing sensitivity or universality. While acceptable for certain applications, these trade-offs hinder the widespread adoption of reagentless biosensing technologies. Here, we report a novel approach to reagentless biosensing that achieves high sensitivity, rapid detection, and universality using the SARS-CoV-2 virus as a model target. Universality is achieved by using nanoscale molecular pendulums, which enables reagentless electrochemical biosensing through a variable antibody recognition element. Enhanced sensitivity and rapid detection are accomplished by incorporating the coffee-ring phenomenon into the sensing scheme, allowing for target preconcentration on a ring-shaped electrode. Using this approach, we obtained limits of detection of 1 fg/mL and 20 copies/mL for the SARS-CoV-2 nucleoproteins and viral particles, respectively. In addition, clinical sample analysis showed excellent agreement with Ct values from PCR-positive SARS-CoV-2 patients.

Despite their unique optical and electrical characteristics, traditional semiconductor quantum dots (QDs) made of heavy metals or carbon are not compatible with many biomedical applications. Cytotoxicity and environmental concerns are key limiting factors that prevent their widescale transition from laboratory research to real-world medical applications. Recently, advanced InP/ZnSe/ZnS QDs have emerged as excellent alternatives to traditional QDs due to their lower toxicity and optical properties; however, they fall short of traditional QDs with respect to their versatility for bioconjugation (i.e., surface chemistry limitations causing unstability in aqueous environments). In this work, we construct a road map for generating, for the first time, highly efficient bio-templated InP/ZnSe/ZnS-aptamers (QDAPT) with long-term stability and high selectivity for applications in targetting bacterial membrane proteins. Our QDAPTs show fast binding reaction kinetics (less than 5 minutes), high brightness, and high shelf-life stability (3 months) after biotemplation in aqueous solvents. We also demonstrate the detection of bacterial membrane proteins on common surfaces using a hand-held imaging device, which attests to the great potential of this system for incorporation into future biomedical technologies.

Nonspecific binding is a significant challenge associated with biosensors in complex food textures. To overcome this, we have developed LISzymes, which are DNAzymes incorporated in lubricant-infused surfaces (LISs). Using milk as a complex background matrix, we show that LISzyme biosensors are significantly more effective in preventing nonspecific binding compared to other commonly used "blocking" methods. The use of lubricant infusion to treat sensing surfaces results in a 4-fold increase in the signal-to-noise ratio obtained with the DNAzyme with respect to untreated surfaces, when detecting the presence of specific bacteria in milk. This is a striking improvement upon previous DNAzyme sensors. We also show that the use of LISs does not affect the DNAzyme's ability to effectively and specifically detect its target─a protein specifically produced by

Portable devices capable of rapid disease detection and health monitoring are crucial to decentralizing diagnostics from clinical laboratories to the patient point-of-need. Although technologies have been developed targeting this challenge, many require the use of reporter molecules or reagents that complicate the automation and autonomy of sensors. New work in the field has targeted reagentless approaches to enable breakthroughs that will allow personalized monitoring of a wide range of biomarkers on demand. This Perspective focuses on the ability of reagentless platforms to revolutionize the field of sensing by allowing rapid and real-time analysis in resource-poor settings. First, we will highlight advantages of reagentless sensing techniques, specifically electrochemical detection strategies. Advances in this field, including the development of wearable and in situ sensors capable of real-time monitoring of biomarkers such as nucleic acids, proteins, viral particles, bacteria, therapeutic agents, and metabolites, will be discussed. Reagentless platforms which allow for wash-free, calibration free-detection with increased dynamic range are highlighted as a key technological advance for autonomous sensing applications. Furthermore, we will highlight remaining challenges which must be overcome to enable widespread use of reagentless devices. Finally, future prospects and potential breakthroughs in precision medicine that will arise as a result of further development of reagentless sensing approaches are discussed.

The development of reagentless sensors that can detect molecular analytes in biological fluids could enable a broad range of applications in personalized health monitoring. However, only a limited set of molecular inputs can currently be detected using reagentless sensors. Here, we report a sensing mechanism that is compatible with the analysis of proteins that are important physiological markers of stress, allergy, cardiovascular health, inflammation and cancer. The sensing method is based on the motion of an inverted molecular pendulum that exhibits field-induced transport modulated by the presence of a bound analyte. We measure the sensor's electric field-mediated transport using the electron-transfer kinetics of an attached reporter molecule. Using time-resolved electrochemical measurements that enable unidirectional motion of our sensor, the presence of an analyte bound to our sensor complex can be tracked continuously in real time. We show that this sensing approach is compatible with making measurements in blood, saliva, urine, tears and sweat and that the sensors can collect data in situ in living animals.

Cross-contamination of biological samples during handling and preparation, is a major issue in laboratory setups, leading to false-positives or false-negatives. Sample carryover residue in pipette tips contributes greatly to this issue. Most pipette tips on the market are manufactured with hydrophobic polymers that are able to repel high surface tension liquids, yet they lack in performance when low surface tension liquids are involved. This presents an obstacle for pipette tips as inaccuracies and loss in precision arise when low surface tension liquids such as viscous oils are pipetted. Here we propose the use of lubricant-infused surface (LIS) technology to achieve omniphobic properties in pipette tips. Using a versatile and simple design, the inner lumen of commercially available pipette tips was coated with a fluorosilane (FS) layer using chemical vapor deposition (CVD). We show that after lubricating the tips through simply pipetting up and down a fluorinated lubricant, the surface free energy of the tips drastically decreased enabling them to attain low retention properties. Contact angle measurements reveals that the treated pipette tips have enhanced omniphobic properties. The repellent behavior of the lubricant-infused pipette tips against physical adsorption is investigated through pipetting a food coloring dye as well as human blood samples and are compared to the untreated tips. The results show significantly less amount carryover residue when the lubricant-infused tips are utilized compared to commercially available ones.

The development of new methods for direct viral detection using streamlined and ideally reagent-free assays is a timely and important, but challenging, problem. The challenge of combatting the COVID-19 pandemic has been exacerbated by the lack of rapid and effective methods to identify viral pathogens like SARS-CoV-2 on-demand. Existing gold standard nucleic acid-based approaches require enzymatic amplification to achieve clinically relevant levels of sensitivity and are not typically used outside of a laboratory setting. Here, we report reagent-free viral sensing that directly reads out the presence of viral particles in 5 minutes using only a sensor-modified electrode chip. The approach relies on a class of electrode-tethered sensors bearing an analyte-binding antibody displayed on a negatively charged DNA linker that also features a tethered redox probe. When a positive potential is applied, the sensor is transported to the electrode surface. Using chronoamperometry, the presence of viral particles and proteins can be detected as these species increase the hydrodynamic drag on the sensor. This report is the first virus-detecting assay that uses the kinetic response of a probe/virus complex to analyze the complexation state of the antibody. We demonstrate the performance of this sensing approach as a means to detect, within 5 min, the presence of the SARS-CoV-2 virus and its associated spike protein in test samples and in unprocessed patient saliva.

The ability to sense biological inputs using self-contained devices unreliant on external reagents or reporters would open countless opportunities to collect information about our health and environment. Currently, a very limited set of molecular inputs can be detected using this type of sensor format. The development of versatile reagentless sensors that could track molecular analytes in biological fluids remains an unmet need. Here, we describe a new universal sensing mechanism that is compatible with the analysis of proteins that are important physiological markers of stress, allergy, cardiovascular health, inflammation and cancer. The sensing mechanism we developed is based on the measurement of field-induced directional diffusion of a nanoscale molecular pendulum tethered to an electrode surface and the sensitivity of electron-transfer reaction kinetics to molecular size. Using time-resolved electrochemical measurements of diffusional motion, the presence of an analyte bound to a sensor complex can be continuously tracked in real time. We show that this sensing approach is compatible with making measurements in blood, saliva, urine, tears and sweat and that the sensors can collect data in situ in living animals. The sensor platform described enables a broad range of applications in personalized health monitoring.

Micropatterned biofunctional surfaces provide a wide range of applications in bioengineering. A key characteristic which is sought in these types of bio-interfaces is prevention of non-specific adhesion for enhanced biofunctionality and targeted binding. Lubricant-infused omniphobic coatings have exhibited superior performance in attenuating non-specific adhesion; however, these coatings completely block the surfaces and do not support targeted adhesion or patterning. In this work, we introduce a novel lubricant-infused surface with biofunctional micropatterned domains integrated within an omniphobic layer. This new class of micropatterned lubricant-infused surfaces simultaneously promotes localized and directed binding of desired targets, as well as repellency of undesired species, especially in human whole blood. Furthermore, this modification method is easily translatable to microfluidic devices offering a wider range of applications and improved performance for immunoassays in whole blood and inhibition of clot formation in microfluidic channels. The biofunctional micropatterned lubricant-infused surfaces were created through a bench-top straight forward process by integrating microcontact printing, chemical vapor deposition (CVD) of self-assembled monolayers (SAMs) of fluorosilanes, and further infusion of the SAMs with a bio-compatible fluorocarbon-based lubricant layer. The developed surfaces, patterned with anti-CD34 antibodies, yield enhanced adhesion and controlled localized binding of target biomolecules (e.g. antibodies) and CD34 positive cells (e.g. HUVECs) inside microfluidic devices, outperforming conventional blocking methods (e.g. bovine serum albumin (BSA) or poly(ethylene glycol) (PEG)) in buffer and human whole blood. These surfaces offer a straightforward and effective way to enhance blocking capabilities while preserving the biofunctionality of a micropatterned system in complex biological environments such as whole blood. We anticipate that these micropatterned biofunctional interfaces will find a wide range of applications in microfluidic devices and biosensors for enhanced and localized targeted binding while preventing non-specific adhesion.

Here, we present a straightforward technique to create bio-functional microfluidic channels using CO2 plasma to induce both carboxylic and hydroxyl groups onto the channel surface. Consequently, not only does the surface allow for irreversible covalent bonding to an oxygen plasma treated PDMS for microfluidic device fabrication, but it also provides functionality for biomolecular immobilization. Furthermore, we demonstrate integration of this technique with microcontact printing to covalently micropattern functional biomolecules inside microfluidic channels. The bio-functionality and efficacy of the microcontact printed antibodies is demonstrated for both bioassays as well as patterning and culturing different cell lines. Results show that the introduced method can be an excellent candidate for cell culture studies in microfluidics. With the new printing method, full cell confluency (∼400 cells per mm2) was achieved after incubation for only 1 day, which is significantly greater than other conventional cell culture techniques inside microfluidic devices. As a proof of concept, we demonstrated the endothelial cells functionality by stimulating von Willebrand Factor secretion under shear stress. This is done via perfusion of histamine through the channel and performing immunofluorescence labeling to observe the inflammatory response of the cells. The developed method eliminates the need for wet chemistry and significantly simplifies producing bio-functional chips which can be used for biosensing, organs-on-chips and tissue engineering applications.

Food safety is a major factor affecting public health and the well-being of society. A possible solution to control food-borne illnesses is through real-time monitoring of the food quality throughout the food supply chain. The development of emerging technologies, such as active and intelligent packaging, has been greatly accelerated in recent years, with a focus on informing consumers about food quality. Advances in the fields of sensors and biosensors has enabled the development of new materials, devices, and multifunctional sensing systems to monitor the quality of food. In this Review, we place the focus on an in-depth summary of the recent technological advances that hold the potential for being incorporated into food packaging to ensure food quality, safety, or monitoring of spoilage. These advanced sensing systems usually target monitoring gas production, humidity, temperature, and microorganisms' growth within packaged food. The implementation of portable and simple-to-use hand-held devices is also discussed in this Review. We highlight the mechanical and optical properties of current materials and systems, along with various limitations associated with each device. The technologies discussed here hold great potential for applications in food packaging and bring us one step closer to enable real-time monitoring of food throughout the supply chain.

This chapter presents various techniques that are employed to engineer and embed biofunctional interfaces in microfluidic platforms. Microfluidic systems exhibit high proficiency in rapid throughput and analysis, as well as small sample and reagent requirements. Achieving biofunctionalization, or controlled immobilization of specific biomolecules, within microchannels is of growing importance due to the rising capabilities of microfluidic systems in biomedical applications including three-dimensional cell culture platforms for tissue- and organ-on-a-chip technologies and point-of-care diagnostics. The formation of cell-functionalized microfluidic devices requires proper microfabrication techniques and biological patterning. Different coating and patterning processes can induce chemically reactive groups on the microchannel’s surface, which can then be used to establish a stable interaction between biomolecules of interest and the surface. The following sections explore widely used approaches to develop cell-laden microchannels: plasma- and silanization-based treatments, microcontact printing, microfluidic patterning, polymer grafting, and extracellular matrix-based hydrogel coating. Furthermore, this chapter introduces numerous surface blocking agents that prevent nonspecific adsorption of biomolecules and boost the sensitivity of biological assays. Through developing target-specific biofunctional interfaces, these techniques enhance the biological compatibility of microfluidic devices for a variety of microfluidic applications such as cell sorting, detection and isolation of circulating tumor cells, and organs-on-chips systems.

Here, we report the development of a transparent, durable, and flexible sensing surface that generates a fluorescence signal in the presence of a specific target bacterium. This material can be used in packaging, and it is capable of monitoring microbial contamination in various types of food products in real time without having to remove the sample or the sensor from the package. The sensor was fabricated by covalently attaching picoliter-sized microarrays of an E. coli-specific RNA-cleaving fluorogenic DNAzyme probe (RFD-EC1) to a thin, flexible, and transparent cyclo-olefin polymer (COP) film. Our experimental results demonstrate that the developed (RFD-EC1)-COP surface is specific, stable for at least 14 days under various pH conditions (pH 3–9), and can detect E. coli in meat and apple juice at concentrations as low as 103 CFU/mL. Furthermore, we demonstrate that our sensor is capable of detecting bacteria while still attached to the food package, which eliminates the need to manipulate the sample. T...

XFe2O4 magnetic nanoparticles (X = Mn, Fe, Co, Ni, and Zn) were prepared by using two methods: coprecipitation and hydrothermal. The synthesized nanoparticles were compared according to the separation in an external magnetic field and finally, the hydrothermal method was specified as a better synthesis method. The magnetic nanoparticles were characterized by physico-chemical analysis methods such as Vibrating Sample Magnetometer (VSM), X-ray diffraction (XRD), Fourier Transform Infrared spectroscopy (FTIR), nitrogen adsorption-adsorption isotherm and Transmission Electron Microscopy (TEM). Magnetic properties of synthesized nanoparticles were studied by ab-initio theoretical methods to confirm and compare with the experimental results. According to the VSM analysis, all of magnetic nanoparticles had good magnetization while CoFe2O4 nanoparticles showed the ferromagnetic behavior. The magnetic properties of XFe2O4 configurations were studied using Density Functional Theory ab-initio method. The theoretical results were consistent with experimental magnetizations in the absence of external field. Finally, the photocatalytic behavior of prepared samples was investigated in the presence of oxone as an accelerated agent for degradation of an azo dye.