Application Of Bioassay Research Articles

PA-Tb-Cu MOF as luminescent nanoenzyme for catalytic assay of hydrogen peroxide

Metal organic frameworks (MOFs) with flexible structures and components have aroused great interest in designing functional materials. In this work, we designed and made a kind of PA-Tb-Cu MOF nanoenzyme capable of emitting fluorescence for the catalytic reaction of hydrogen peroxide (H2O2). Luminescent Tb3+, catalytic Cu2+ and bridging ligand were assembled and integrated into a single material nanoenzyme. This PA-Tb-Cu MOF nanoenzyme not only possessed excellent catalytic activity comparable to horseradish peroxidase but also can real-time fluorescently indicate the concentration of H2O2 as low as 0.2µM during catalysis. Luminescent PA-Tb-Cu MOF nanoenzyme did not need a common combined use of natural/artificial enzymes and chromogenic reactions for the quantification of H2O2 in widely-used enzyme-catalytic reactions. The present strategy assembled directly from functional ions/molecules provides a new way for the design and development of smart, multifunctional artificial enzymes for wide applications in biocatalysis, bioassays and nano-biomedicine.

Fluorescence and Scattering Light Cross Correlation Spectroscopy and Its Applications in Homogeneous Immunoassay.

In this work, we propose fluorescence and scattering light cross-correlation spectroscopy (FSCCS) based on laser confocal configuration using silver nanoparticle (SNPs) and Alexa Fluor 488 (Alexa) as probe pairs. FSCCS is a single molecule (particle) method, and its principle is similar to that of fluorescence cross-correlation spectroscopy (FCCS). We established the setup of FSCCS using single wavelength laser and developed an immunoassay model of FSCCS. The reliability and adaptability of FSCCS method were evaluated by homogeneous sandwich immunoassay mode. In the study, liver cancer biomarker alpha-fetoprotein (AFP) was used as an assay model, two different antibodies were labeled with SNPs and fluorophore Alexa Fluor 488, respectively. In the optimal conditions, the linear range of AFP covers 5 pM to 580 pM and the detection limit is 3.1 pM. This method was successfully applied for direct determination of AFP levels in human serum samples, and the obtained results were in good agreement with data obtained via ELISAs. The advantage of this method lies in its simplicity, attractive SNPs probes, high sensitivity and selectivity and high efficiency. We believe that FSCCS method exhibits promising potential applications in homogeneous bioassays and study on the molecular interaction and nanoparticle-molecule interaction.

Based on DNA Strand Displacement and Functionalized Magnetic Nanoparticles: A Promising Strategy for Enzyme Immobilization

The efforts to enhance the stability of enzymes and improve their activity have generated considerable interest because of their wide applications in bioenergy conversion, proteomics research, and bioassays. This study reports a promising strategy for enzyme immobilization based on toehold-mediated DNA strand displacement on functionalized magnetic nanoparticles for the first time. The strategy provided a convenient approach to achieve sequential displacement and immobilization of different enzymes, using alkaline phosphatase (ALP), horseradish peroxidase (HRP), and trypsin as different model enzymes. Taking trypsin as an example, the enzyme immobilization procedure by DNA strand displacement exhibited high reversibility and reproducibility, which could retrieve more than 87% of the enzymatic activity after consecutive hybridization and dehybridization cycles. The thermal stability of the immobilized trypsin was significantly enhanced up to 3.1- and 2.3-fold greater than free enzyme after 45 min incubatio...

Cooking Time and Iron Bioavailability: The USDA‐ARS Approach to Bean Fe Biofortification

Beans (Phaseolis vulgaris L.) represent a significant portion of the diet for many populations at risk of Fe deficiency and are therefore a crop targeted for Fe biofortification. In Sub-Saharan Africa, 75% of the population use wood or charcoal as the primary cooking fuel; thus, the expense of cooking fuel factors heavily in selection of beans by consumers in this region prone to Fe deficiency. Our approach to bean Fe biofortification incorporates identifying and developing lines that are fast-cooking, acceptable to farmers and consumers, high in overall nutritional quality, and specifically high in bioavailable or bioaccessible (ie. “absorbable”) Fe as measured by an established Caco-2 cell bioassay. This approach is significant as this bioassay has now been thoroughly validated against animal studies and human efficacy studies of Fe biofortified crops. Application of the Caco-2 bioassay has recently resulted in the development of a Health and Nutrition Panel (HNP) to demonstrate the above approach. The HNP is composed of 12 cultivars refined from the screening of over 200 accessions from the Andean Diversity Panel. The HNP represents a defined set of lines that demonstrates how the fast-cooking trait can be associated with increased nutrition via increased bean consumption and via improved Fe bioaccessibility. The fast-cooking trait is highly tractable across multiple locations and harvest years. Fast cooking lines have the potential to save several hours of time and fuel. Such savings may allow for purchase of greater quantities of beans and thus increase bean consumption. As added benefit, increased Fe bioaccessibility also appears to be associated with the fast –cooking trait, and is particularly high in certain yellow bean varieties. Additional studies have shown that in direct comparison to high Fe lines of beans, some fast cooking yellow beans appear to deliver 2–3 times more absorbable Fe, even though they have lesser amounts of Fe. It is important to note that some of these fast cooking lines are already present in the African marketplace and are highly desired by consumers; however, they are not yet recognized for their nutrition potential. It is the goal of this program to fully characterize these lines, develop and continue breeding to enhance the fast-cooking and nutritional traits, and educate consumers of the nutritional benefits. Support or Funding InformationThis research funded by USDA-ARS. The Health and Nutrition Panel

Toxicological impacts of antibiotics on aquatic micro-organisms: A mini-review.

Antibiotics are found globally in the environment at trace levels due to their extensive consumption, which raises concerns about the effects they can have on non-target organisms, especially environmental micro-organisms. So far the majority of studies have focused on different aspects of antibiotic resistance or on analyzing the occurrence, fate, and removal of antibiotics from hospital and municipal wastewaters. Little attention has been paid to ecotoxicological effects of antibiotics on aquatic micro-organisms although they play a critical role in most ecosystems and they are potentially sensitive to these substances. Here we review the current state of research on the toxicological impacts of antibiotics to aquatic micro-organisms, including proteobacteria, cyanobacteria, algae and bacteria commonly present in biological wastewater treatment processes. We focus on antibiotics that are poorly removed during wastewater treatment and thus end up in surface waters. We critically discuss and compare the available analytical methods and test organisms based on effect concentrations and identify the knowledge gaps and future challenges. We conclude that, in general, cyanobacteria and ammonium oxidizing bacteria are the most sensitive micro-organisms to antibiotics. It is important to include chronic tests in ecotoxicological assessment, because acute tests are not always appropriate in case of low sensitivity (for example for proteobacteria). However, the issue of rapid development of antibiotic resistance should be regarded in chronic testing. Furthermore, the application of other species of bacteria and endpoints should be considered in the future, not forgetting the mixture effect and bacterial community studies. Due to differences in the sensitivity of different test organisms to individual antibiotic substances, the application of several bioassays with varying test organisms would provide more comprehensive data for the risk assessment of antibiotics. Regardless of the growing concerns related to antibiotics in the environment, there are still evident knowledge gaps related to antibiotics, as there is only limited or no ecotoxicological data on many potentially harmful antibiotics.

Synthesis of the Src SH2 domain and its application in bioassays for mirror-image screening

Mirror-image screening systems for Src SH2 domain inhibitors were established using a synthetic Src SH2 domain.

Motor-assisted chip-in-a-tube (MACT): a new 2- and 3-dimensional centrifugal microfluidic platform for biomedical applications.

Currently, centrifuge apparatus is primarily an end-point sample processing piece of equipment. The lack of real-time active control has imposed an inherent limitation such that many delicate sample processing steps requiring immediate and accurate intervention have never been possible. We report herein a motor-assisted chip-in-a-tube (MACT) platform in which a microfluidic chip placed inside a common centrifuge canister can be rotated through wireless control in order to manipulate the centrifugal force vector in a 3-dimensional (3D) manner. As a demonstration experiment, we have used our MACT prototype to perform the operation for two common biomedical procedures, namely human blood plasma separation and E. coli plasmid DNA extraction. This simple, yet highly effective and versatile approach may serve as a generic one-for-all platform for a wide range of common laboratory experiments and bioassay applications.

Reproducible fiber optofluidic laser for disposable and array applications.

Disposable sensors are widely used in biomedical detection due to their inherent safety, ease of use and low cost. An optofluidic laser is a sensitive bioassay platform; however, demonstrating its fabrication cheaply and reproducibly enough for disposable use has been challenging. Here, we report a low-cost, reproducible fiber optofluidic laser (FOFL) using a microstructured optical fiber (MOF). The MOF not only supports the whispering gallery modes for lasing but also serves as a microfluidic channel for sampling the liquid gain medium via capillary force. Because of the precise control of its geometry (δ < 0.4%) during the fiber-drawing process, good reproducibility in laser intensity (δ = 6.5%) was demonstrated by changing 10 sections of the MOF. The strong coupling between the in-fiber resonator and gain medium enables a low threshold of 3.2 μJ mm-2. The angular dependence of the laser emission was observed experimentally and analyzed with numerical simulations. An array of the FOFLs was also demonstrated. This technology has great potential for low-cost bioassay applications.

Plasmon-enhanced fluorescence imaging with silicon-based silver chips for protein and nucleic acid assay

Metal-enhanced fluorescence shows great potential for improving the sensitivity of fluoroscopy, which has been widely used in protein and nucleic acid detection for biosensor and bioassay applications. In comparison with the traditional glass-supported metal nanoparticles (MNPs), the introduction of a silicon substrate has been shown to provide an increased surface-enhanced Raman scattering (SERS) effect due to the coupling between the MNPs and the semiconducting silicon substrate. In this work, we further study the fluorescence-enhanced effect of the silicon-supported silver-island (Ag@Si) plasmonic chips. In particular, we investigate their practical application of improving the traditional immunoassay such as the biotin-streptavidin-based protein assay and the protein-/nucleic acid-labeled cell and tissue samples. The protein assay shows a wavelength-dependent enhancement effect of the Ag@Si chip, with an enhancement factor ranging from 1.2 (at 532 nm) to 57.3 (at 800 nm). Moreover, for the protein- and nucleic acid-labeled cell and tissue samples, the Ag@Si chip provides a fluorescence enhancement factor of 3.0–4.1 (at 800 nm) and a significant improvement in the signal/background ratio for the microscopy images. Such a ready accommodation of the fluorescence-enhanced effect for the immunoassay samples with simple manipulations indicates broad potential for applications of the Ag@Si chip not only in biological studies but also in the clinical field.

Fluorescent Detection of Phosphate in Aqueous Solution Based on Near Infrared Emission Ag2S QDs/Metal-Organic Shell Composite.

A novel fluorescence method for sensitive and selective detection of phosphate was developed based on near infrared emission Ag2S QDs/ Metal-Organic Shell Composite via the deposition of metal-organic (zinc-nitrogen) coordination shell around Ag2S QDs . Under optimal conditions described, the fluorescence intensity of the composite was decreased at 685nm in the presence of phosphate, which was linearly related to the concentration of phosphate in the range of 0. 7 to 4.2μM and 11.2 to 88.2μM with the relative correlation coefficient of R2=0.998 and 0.987 respectively and detection limit as low as 6nM. In addition, the proposed method was successfully utilized in serum samples, tap water and Yangtze River water samples with the recoveries ranged from 94.76 to 100.86%, which presaged more opportunities for application in related bioassay and water sample researches.

Magnetic/Fluorescent Barcodes Based on Cadmium‐Free Near‐Infrared‐Emitting Quantum Dots for Multiplexed Detection

Magnetic/fluorescent barcodes, which combine quantum dots (QDs) and superparamagnetic nanoparticles in micrometer‐sized host microspheres, are promising for automatic high‐throughput multiplexed biodetection applications and “point of care” biodetection. However, the fluorescence intensity of QDs sharply decreases after addition of magnetic nanoparticles (MNPs) due to absorption by MNPs, and thus, the encoding capacity of QDs becomes more limited. Furthermore, the intrinsic toxicity of cadmium‐based QDs, the most commonly used QD in barcodes, has significant risks to human health and the environment. In this work, to alleviate fluorescence quenching and intrinsic toxicity, cadmium‐free NIR‐emitting CuInS2/ZnS QDs and Fe3O4MNPs are successfully incorporated into poly(styrene‐co‐maleic anhydride) microspheres by using the Shirasu porous glass membrane emulsification technique. A “single‐wavelength” encoding model is successfully constructed to guide the encoding of NIR QDs with wide emission spectra. Then, a “single‐wavelength” encoding combined with size encoding is used to produce different optical codes by simply changing the wavelength and the intensity of the QDs as well as the size of the barcode microspheres. 48 barcodes are easily created due to the greatly reduced energy transfer between the NIR‐emitting QDs and MNPs. The resulting bifunctional barcodes are also combined with a flow cytometer using one laser for multiplexed detection of five tumor markers in one test. Assays based on these barcodes are significantly more sensitive than non‐magnetic and traditional ELISA assays. Moreover, validating experiments also show good performance of the bifunctional barcodes‐based suspension array when dealing with patient serum samples. Thus, magnetic/fluorescent barcodes based on NIR‐emitting CuInS2/ZnS QDs are promising for multiplexed bioassay applications.

Novel hybrid probe based on double recognition of aptamer-molecularly imprinted polymer grafted on upconversion nanoparticles for enrofloxacin sensing

A novel luminescent “double recognition” method for the detection of enrofloxacin (ENR) is developed to overcome some of the challenges faced by conventional molecularly imprinting. Biotinylated ENR aptamers immobilised on upconversion nanoparticles (UCNPs) surface are implemented to capture and concentrate ENR as the first imprinting recognition safeguard. After correct folding of the aptamer upon the existing targets, polymerization of methacrylic acid monomers around the ENR-aptamer complexes to interact with the residual functional groups of ENR by using molecularly imprinting techniques is the second imprinting recognition safeguard. The “double recognition” imprinting cavities are formed after removal of ENR, displaying recognition properties superior to that of aptamer or traditional molecularly imprinting alone. Another interest of this method is simultaneous molecular recognition and signal conversion by fabricating the “double recognition” receptor on to the surface of UCNPs to form a hybrid sensing system of apta-MIP/UCNPs. The proposed sensing method is applied to measure ENR in different fish samples. Good recoveries between 87.05% and 96.24%, and relative standard deviation (RSD) values in the range of 1.19–4.83% are obtained, with the limits of detection and quantification of 0.04 and 0.12ng/mL, respectively. It indicates that the sensing method is feasible for the quantification of target ENRs in real samples, and show great potential for wide-ranging application in bioassays.

Ultrasensitive sandwich-type electrochemical immunosensor based on trimetallic nanocomposite signal amplification strategy for the ultrasensitive detection of CEA.

A novel and ultrasensitive sandwich-type electrochemical immunosensor was designed for the quantitative detection of carcino-embryonic antigen (CEA). This immunosensor was developed by using the trimetallic NiAuPt nanoparticles on graphene nanosheets (NGs) nanosheets (NiAuPt-NGs) as excellent labels and β-cyclodextrin functionalized reduced graphene oxide nanosheets (CD-NGs) as the platform. The CD-NGs with high specific surface area good biocompatibility and the ideal dispersibility was used to capture the primary antibodies (Ab1) efficiently. The trimetallic NiAuPt-NGs nanocomposites were used as the labels for signal amplification, showing better electrocatalytic activity towards the reduction of hydrogen peroxide (H2O2), which is much better than that the monometallic Pt-NGs, bimetallic NiPt-NGs and AuPt-NGs due to the synergetic effect presented in NiAuPt-NGs. The NiAuPt-NGs nanocomposites consist of tightly coupled nanostructures of Au, Ni and Pt, which have neither an alloy nor a core-shell structure. Under the optimal conditions, a linear range from 0.001–100 ng/mL and a low detection limit of 0.27 pg/mL were obtained for CEA. The proposed electrochemical sandwich-type immunosensor may have a promising application in bioassay and it enriches the electrochemical immunoassays.

A paper/polymer hybrid microfluidic microplate for rapid quantitative detection of multiple disease biomarkers.

Enzyme linked immunosorbent assay (ELISA) is one of the most widely used laboratory disease diagnosis methods. However, performing ELISA in low-resource settings is limited by long incubation time, large volumes of precious reagents, and well-equipped laboratories. Herein, we developed a simple, miniaturized paper/PMMA (poly(methyl methacrylate)) hybrid microfluidic microplate for low-cost, high throughput, and point-of-care (POC) infectious disease diagnosis. The novel use of porous paper in flow-through microwells facilitates rapid antibody/antigen immobilization and efficient washing, avoiding complicated surface modifications. The top reagent delivery channels can simply transfer reagents to multiple microwells thus avoiding repeated manual pipetting and costly robots. Results of colorimetric ELISA can be observed within an hour by the naked eye. Quantitative analysis was achieved by calculating the brightness of images scanned by an office scanner. Immunoglobulin G (IgG) and Hepatitis B surface Antigen (HBsAg) were quantitatively analyzed with good reliability in human serum samples. Without using any specialized equipment, the limits of detection of 1.6 ng/mL for IgG and 1.3 ng/mL for HBsAg were achieved, which were comparable to commercial ELISA kits using specialized equipment. We envisage that this simple POC hybrid microplate can have broad applications in various bioassays, especially in resource-limited settings.

Compact Nanowire Sensors Probe Microdroplets

The conjunction of miniature nanosensors and droplet-based microfluidic systems conceptually opens a new route toward sensitive, optics-less analysis of biochemical processes with high throughput, where a single device can be employed for probing of thousands of independent reactors. Here we combine droplet microfluidics with the compact silicon nanowire based field effect transistor (SiNW FET) for in-flow electrical detection of aqueous droplets one by one. We chemically probe the content of numerous (∼10(4)) droplets as independent events and resolve the pH values and ionic strengths of the encapsulated solution, resulting in a change of the source-drain current ISD through the nanowires. Further, we discuss the specificities of emulsion sensing using ion sensitive FETs and study the effect of droplet sizes with respect to the sensor area, as well as its role on the ability to sense the interior of the aqueous reservoir. Finally, we demonstrate the capability of the novel droplets based nanowire platform for bioassay applications and carry out a glucose oxidase (GOx) enzymatic test for glucose detection, providing also the reference readout with an integrated parallel optical detector.

On the Use of Scalable NanoISFET Arrays of Silicon with Highly Reproducible Sensor Performance for Biosensor Applications.

As a prerequisite to the development of real label-free bioassay applications, a high-throughput top–down nanofabrication process is carried out with a combination of nanoimprint lithography, anisotropic wet-etching, and photolithography methods realizing nanoISFET arrays that are then analyzed for identical sensor characteristics. Here, a newly designed array-based sensor chip exhibits 32 high aspect ratio silicon nanowires (SiNWs) laid out in parallel with 8 unit groups that are connected to a very highly doped, Π-shaped common source and individual drain contacts. Intricately designed contact lines exert equal feed-line resistances and capacitances to homogenize the sensor response as well as to minimize parasitic transport effects and to render easy integration of a fluidic layer on top. The scalable nanofabrication process as outlined in this article casts out a total of 2496 nanowires (NWs) on a 4 inch p-type silicon-on-insulator (SOI) wafer, yielding 78 sensor chips based on nanoISFET arrays. The sensor platform exhibiting high-performance transistor characteristics in buffer solutions is thoroughly characterized using state-of-the-art surface and electrical measurement techniques. Deploying a pH sensor in liquid buffers after high-quality gas-phase silanization, nanoISEFT arrays demonstrate typical pH sensor behavior with sensitivity as high as 43 ± 3 mV·pH–1 and a device-to-device variation of 7% at the wafer scale. Demonstration of a high-density sensor platform with uniform characteristics such as nanoISFET arrays of silicon (Si) in a routine and refined nanofabrication process may serve as an ideal solution deployable for real assay-based applications.

Electrochemical immunosensor for detection of prostate specific antigen based on an acid cleavable linker into MSN-based controlled release system

A mesoporous silica nanoparticle (MSN)-based controlled release system with acid cleavable linkage was developed to fabricate an electrochemical immunosensor for the quantitative detection of the prostate-specific antigen (PSA). 3,9-Bis(3-aminopropyl)-2,4,8,10-tetraoxaspiro[5.5]undecane functionalized mesoporous silica nanoparticles (MSN-Acetal) were used to immobilize the electron mediator thionine (Th). The encapsulation of Th molecules was achieved by capping the pores of MSN-Acetal with carboxylic acid modified Au nanoparticles (defined as MSN-Th-Au). Under the acidic conditions, the capped Au nanoparticles were removed from MSN-Th-Au through the hydrolysis of the acid-labile acetal linker, resulting in the release of encapsulated Th. In this work, the pH-responsive cargo release system was firstly used as the label of secondary anti-PSA for developing an electrochemical immunosensor, and amination Fe3O4 was used as the sensing matrix for immobilizing primary anti-PSA on magnetic carbon electrode surfaces. The specific recognition of PSA resulted in the attachment of MSN-Th-Au-secondary anti-PSA (MSN-Th-Au-Ab2) onto the electrode surfaces. Subsequently, the released Th was detected by differential pulse voltammetry under the acidic conditions. The developed cargo release system provided an innovative and reliable method for the detection of PSA because the response signal was correlated with the concentration of PSA. Under the optimal conditions, the electrochemical immunosensor exhibited a wide linear range of 0.001–5.0ng/mL with a low detection limit of 0.31pg/mL. Moreover, the developed immunosensor showed superior reproducibility and long-term stability, which has promising applications in bioassay and biosensing.

Modification on populating paths of β-NaYF_4:Nd/Yb/Ho@SiO_2@Ag core/double-shell nanocomposites with plasmon enhanced upconversion emission

Nd/Yb/Ho-doped hexagonal NaYF4 nanocrystals were synthesized and excited at 808 nm to avoid an overheating effect under 980 nm laser excitation. A core/double-shell nanocomposite (NaYF4:Nd/Yb/Ho@SiO2@Ag nanoparticles) was then developed to enhance upconversion luminescence intensity. The fabricated system exhibited a metal-induced effect and was used to determine the influence of SiO2 layer thickness on upconverting optical properties. SiO2 layer with different thicknesses were used to adjust the distance between NaYF4 nanocrystals and the outer layer of Ag nanocrystals. The largest enhancement of upconversion luminescence occurred when 10 nm-thick SiO2 shell was used, with enhancement factors of 15 and 7.5 for green and red emissions, respectively. Results indicated that the degree of luminescence enhancement was correlated with the competition of the three interactions occurred between fluorophores and metal NPs. After coating with Ag nanoparticles, some modifications were found on the upconversion processes of NaYF4 core nanoparticles and the underlying mechanisms were discussed respectively. The controllable upconversion luminescence renders metal-upconverted nanocomposites as suitable for bioassay, bio-imaging, and energy conversion applications, which require high sensitivity and low background.

Facile and High-Throughput Synthesis of Functional Microparticles with Quick Response Codes.

Encoded microparticles are high demand in multiplexed assays and labeling. However, the current methods for the synthesis and coding of microparticles either lack robustness and reliability, or possess limited coding capacity. Here, a massive coding of dissociated elements (MiCODE) technology based on innovation of a chemically reactive off-stoichimetry thiol-allyl photocurable polymer and standard lithography to produce a large number of quick response (QR) code microparticles is introduced. The coding process is performed by photobleaching the QR code patterns on microparticles when fluorophores are incorporated into the prepolymer formulation. The fabricated encoded microparticles can be released from a substrate without changing their features. Excess thiol functionality on the microparticle surface allows for grafting of amine groups and further DNA probes. A multiplexed assay is demonstrated using the DNA-grafted QR code microparticles. The MiCODE technology is further characterized by showing the incorporation of BODIPY-maleimide (BDP-M) and Nile Red fluorophores for coding and the use of microcontact printing for immobilizing DNA probes on microparticle surfaces. This versatile technology leverages mature lithography facilities for fabrication and thus is amenable to scale-up in the future, with potential applications in bioassays and in labeling consumer products.

Optimization of a Liquid Crystal‐based Sensory Platform for Monitoring Enzymatic Glucose Oxidation

Managing glucose levels in human blood is extremely important for the treatment of diabetes. Here, an innovative sensory strategy has been developed to monitor the enzymatic activities of glucose and glucose oxidase by using confined liquid crystal (LC) birefringent droplet patterns. Acidic products released during the glucose oxidation process lead to a slight decrease in the pH of aqueous systems that can be monitored by pH‐sensitive LC materials. Of the existing pH‐sensitive LC materials, dodecanoic acid‐doped 4‐cyano‐4′‐pentylbiphenyl is inexpensive and easily adjusted to satisfy the 7.4 ± 0.05 pH requirement of human blood. Moreover, the orientational alignment of capillary‐confined pH‐responsive LCs can be disrupted at the aqueous/LC interface following a slight decrease in the critical pH of aqueous reaction systems, which results in an optical signal that can be observed with the naked eye by using polarizing optical microscopy. Based on the stable LC droplet patterns generated by the cylindrical confinement system, the functionalized LCs can selectively detect glucose at concentrations as low as 0.1 pM. This study further advances the previously reported LC‐based glucose monitoring systems by reducing production costs and instituting a smarter LC sensory design. This improved system shows potential for the use in clinical bioassay applications.