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In recent years, there has been increasing demand for miniaturized analytical instruments that allow immediate on-site analysis. We have been developing a portable HPLC system consisting of a column-integrated chip and a compact, low-power electroosmotic flow pump. This study aims to extend the versatility of the portable HPLC system with a further reduction of the overall system size, and thus we constructed a UV absorbance detection module that can be mounted to the column-integrated chip. For this detection module, a UV light-emitting diode (peak wavelength = 255 nm) and photodiode were used as a light source and photodetector, and a system to control their operation and process the signals was also constructed. The developed detection module exhibited excellent performances as an HPLC detector., 近年，サンプリングした現場で即時に分析が可能な小型分析装置の需要が高まっている．著者らは，小型・低電力の電気浸透流ポンプとカラム内蔵チップからなる小型HPLCシステムの開発に取り組んできた．本研究では，小型HPLCシステム全体のさらなる小型化を目指しながら検出における汎用性を拡張するため，カラム内蔵チップに適用可能で，汎用性が高い紫外吸光度検出モジュールを構築した．そのために光源及び受光器に深紫外発光ダイオード（ピーク発光波長255 nm）及びフォトダイオードを用いるとともに，これらの動作を制御し，信号を処理するシステムも構築した．さらに，検出モジュールが市販分光光度計並みの感度を持ち，良好にクロマトグラム測定ができることを実証した．

The translation of nanoparticles (NPs) from laboratory to clinical settings is limited, which is not ideal. One of the reasons for this is that we currently have limited ability to precisely regulate various physicochemical parameters of nanoparticles. This has made it difficult to rapidly perform targeted screening of drug preparation conditions. In this study, we attempted to broaden the range of preparation conditions for particle size-modulated poly(lactic-co-glycolic-acid) (PLGA) NP to enhance their applicability for drug delivery systems (DDS). This was done using a variety of organic solvents and a glass-based microfluidic device. Furthermore, we compared the PDMS-based microfluidic device to the glass-based microfluidic device in terms of the possibility of a wider range of preparation conditions, especially the effect of different solvents on the size of the PLGA NPs. PLGA NPs with different sizes (sub-200 nm) were successfully prepared, and three different types of taxanes were employed for encapsulation. The drug-loaded NPs showed size-dependent cytotoxicity in cellular assays, regardless of the taxane drug used.

The majority of chemical experiments conducted during educational programs are carried out in laboratories because they require instructors with special skills, in addition to large, expensive instruments. Recently, there has been an increasing demand for chemical experiments that can be carried out anywhere. Herein, we propose a novel type of paper-based analytical device (PAD) for enabling quantitative analysis without the requirement for a micropipette, since the PAD features a large sample loading zone and a waste zone, enabling accurate volume control of a liquid sample. We initially employed a micropipette to evaluate the PAD and demonstrate the quantitative analysis of ascorbic acid (AA) and pH using different loading volumes. Finally, we determined the AA concentrations and pH values of commercially available beverages using disposable plastic droppers, and the obtained results were in good agreement with those obtained through conventional methods. This PAD format can therefore be used as a novel educational tool for conducting certain chemical analyses in remote learning environments.

Microfluidic devices are widely used in lipid nanoparticle (LNP)-based vaccines and nanomedicine research. These devices should be stiff enough to withstand the high flow rate for the mass production of LNPs, and malleable enough to use when fabricating complicated microchannel or micromixer structures, such as stag-gering herringbone micromixers. Due to the limitations of the available fabrication methods, optimal micro -fluidic devices have not yet been developed. In this study, we report the development of a glass-based microfluidic device based on the invasive Lipid Nanoparticle Production (iLiNP) device (R) reported previously. The LNP size controllability of glass-based iLiNP device was similar to that of the poly(dimethylsiloxane) (PDMS)-based iLiNP device, and the glass-iLiNP device was used for mRNA-loaded LNP production with ionizable lipids used for COVID-19 mRNA vaccines. We also demonstrate a piling-and numbering-up strategy based on glass-iLiNP device. The iLiNP unit composed of five-layered microchannels was fabricated by piling-up each glass-iLiNP device followed by parallelization (numbering-up) for the mass production of LNPs. This iLiNP system can produce LNPs with sizes ranging between 20 and 60 nm at a flow rate of 20-50 mL/min, and its performance is comparable to that of the commercially available microfluidic system like NanoAssemblr (R).

Analytical methods with fluorescence detection are in widespread use for detecting low abundance analytes. Here, we report a simple method for fluorescence signal amplification utilizing a structure of an azide-unit pendant water-soluble photopolymer (AWP) in a microchannel. The AWP is a poly(vinyl alcohol)-based photocross-linkable polymer, which is often used in biosensors. We determined that the wall-like structure of the AWP (AWP-wall) constructed in a microchannel functioned as an amplifier of a fluorescence signal. When a solution of fluorescent molecules was introduced into the microchannel having the AWP-wall, the fluorescent molecules accumulated inside the AWP-wall by diffusion. Consequently, the fluorescence intensity inside the AWP-wall increased locally. Among the fluorescent molecules considered in this paper, 9H-(1,3-dichloro-9,9-dimethylacridin-2-one-7-yl) (DDAO) showed the highest efficiency of fluorescence signal amplification. We prepared a calibration curve for DDAO using the fluorescence intensity inside the AWP-wall, and the sensitivity was 5-fold that for the microchannel without the AWP-wall. This method realizes the improved sensitivity of fluorescence detection easily because the fluorescence signal was amplified only by injecting the solution into the microchannel having the AWP-wall. Furthermore, since this method is not limited to only the use of microchannel, we expect it to be applicable in various fields.

The development of low-cost, user-friendly paper-based analytical devices (PADs) that can easily measure target chemicals is attracting attention. However, most PADs require manipulation of the sample using sophisticated micropipettes for quantitative analyses, which restricts their user-friendliness. In addition, immobilization of detection molecules to cellulose fibers is essential for achieving good measuring ability as it ensures the homogeneity of color development. Here, we have described a dip-type PAD that does not require pipette manipulation for sample introduction and immobilization of detection molecules to cellulose fibers and its application to ascorbic acid (AA) and pH assays. The PAD consisted of a dipping area and two channels, each with two detection zones. The developed PADs show color distribution in the two detection zones depending on the sample flow from the dipping area. In comparison with a PAD that has one detection zone at the end of the channel, our developed device achieved higher sensitivity (limit of detection (LOD), 0.22 mg/mL) and reproducibility (maximum coefficient of variation (CV), 2.4%) in AA detection. However, in pH detection, the reproducibility of the PAD with one detection zone at the end of the channel (maximum CV, 21%) was worse than that with two zones (maximum CV, 11%). Furthermore, a dipping time over 3 s did not affect color formation or calibration curves in AA detection: LODs at 3 and 30 s dipping time were 18 and 5.8 μg/mL, respectively. The simultaneous determination of AA and pH in various beverages was performed with no significant difference compared to results of the conventional method.

Nowadays, the diagnosis of viral infections is receiving broad attention. We have developed a non-competitive fluorescence polarization immunoassay (NC-FPIA), which is a separation-free immunoassay, for a virus detection. H5 subtype avian influenza virus (H5-AIV) was used as a model virus for the proof of concept. The fluorescein-labeled Fab fragment that binds to H5 hemagglutinin was used for NC-FPIA. The purified H5-AIV which has H5 hemagglutinin was mixed with the fluorescein-labeled Fab fragment. After that, the degree of fluorescence polarization was measured with a portable FPIA analyzer. H5-AIV was successfully detected with an incubation time of 15 min. In addition, the portable FPIA analyzer enables performance of on-site NC-FPIA with a sample volume of 20 μL or less. This is the first research of detecting a virus particle by FPIA. This NC-FPIA can be applied to rapid on-site diagnosis of various viruses. Supplementary Information The online version contains supplementary material available at 10.1007/s00216-021-03193-y.

Competitive immunoassays comprise the standard means of detecting small molecules. However, conventional methods using microwells are difficult to apply during point-of-care tests (POCT) because they require complicated handling and are time consuming. Although paper-based analytical devices (PAD) have received considerable focus because of their rapid and straightforward operation, only a few devices have been proposed for competitive immunoassays. Herein, we describe a novel universal PAD format with a 3-dimensional configuration for competitive immunoassays that rapidly and sensitively detects small molecules. The proposed device comprised a layered structure with uniform color formation and high capture efficiency between antigen and antibody that results in rapid and reproducible results. The device rapidly (90 s) assayed biotin as a model target, with a limit of detection (LOD) of 5.08 ng mL−1, and detected progesterone with an LOD of 84 pg mL−1 within 5 min. Moreover, sample volumes and reagent consumption rates were minimized. Thus, our device could be applied to competitive immunoassays of various small molecules in POCT.

Size-controlled lipid nanoparticle (LNP)-based DNA/RNA delivery is a leading technology for gene therapies. For DNA/RNA delivery, typically, a cationic lipid is used to encapsulate DNA/RNA into LNPs. However, the use of the cationic lipid leads to cytotoxicity. In contrast, noncationic NPs, such as exosomes, are ideal nanocarriers for DNA/RNA delivery. However, the development of a simple one-step method for the production of size-controlled noncationic NPs encapsulating DNA/RNA is still challenging because of the lack of electrostatic interactions between the cationic lipid and negatively charged DNA/RNA. Herein, we report a microfluidic-based one-step method for the production of size-controlled noncationic NPs encapsulating small interfering RNA (siRNA). Our microfluidic device, named iLiNP, enables the efficient encapsulation of siRNA, as well as control over the NP size, by varying the flow conditions. We applied this method to produce size-controlled exosome-like NPs. The siRNA-loaded exosome-like NPs did not show in vitro cytotoxicity at a high siRNA dosage. In addition, we investigated the effect of the size of the exosome-like NPs on the target gene silencing and found that the 40-50 nm-sized NPs suppressed target protein expression at a dose of 20 nM siRNA. The iLiNP-based one-step production method for size-controlled noncationic-NP-encapsulated RNA is a promising method for the production of artificial exosomes and functionally modified exosomes for gene and cell therapies.

This paper is the first report of a non-competitive fluorescence polarization immunoassay (NC-FPIA) using a peptide as a tracer. The NC-FPIA can easily and quickly quantify the target after simply mixing them together. This feature is desirable for point-of-need applications such as clinical diagnostics, infectious disease screening, on-site analysis for food safety

Acute physiological changes after balloon angioplasty are very important because of acute recoil and dissection. However, serial physiological assessments after drug-coated balloons (DCB) have not been investigated.This prospective observational single-center study evaluated 50 lesions that underwent optical coherence tomography (OCT)-guided percutaneous coronary intervention (PCI) with DCB and a 9-months angiographical and OCT follow-up. Fractional flow reserve (FFR) and instantaneous wave-free ratio (iFR) were measured immediately (FFR0m and, iFR0m) and 15 min (FFR15m and iFR15m) after DCB, and the difference (dif-FFR and, dif-iFR) was calculated. The iFR gradients during lesions treated with DCB were measured. For OCT and quantitative coronary angiography (QCA) data, delta values were calculated.At index PCI, three lesions were needed for bailout stenting. At follow-up, 47 lesions were divided into two groups according to the delta minimal lumen area (MLA) on OCT: late lumen enlargement (LLE) (n = 29) and non-LLE (n = 18). In LLE group, FFR15m and iFR15m were significantly high (0.90 ± 0.03 vs. 0.85 ± 0.07, p 0.001, 0.97 ± 0.02 vs. 0.92 ± 0.10, p = 0.008, respectively) and %AS on OCT, dif-FFR and dif-iFR were significantly low (38.5% (33.6, 42.3) vs. 45.1% (38.9, 54.6), p = 0.03, -0.001 ± 0.006 vs. 0.036 ± 0.032, p 0.001, -0.002 ± 0.008 vs. 0.019 ± 0.017, p 0.001, respectively) compared with non-LLE group. However, there were no significant differences in FFR0m, iFR0m, or any other OCT or QCA data.Post-physiological assessment and a decrease in physiological indices in the first 15 min after PCI are important for treating de novo lesions using the DCB strategy.

The development of functional lipid nanoparticles (LNPs) is one of the major challenges in the field of drug delivery systems (DDS). Recently, LNP-based RNA delivery systems, namely, RNA-loaded LNPs have attracted attention for RNA therapy. In particular, mRNA-loaded LNP vaccines were approved to prevent COVID-19, thereby leading to the paradigm shift toward the development of next-generation nanomedicines. For the LNP-based nanomedicines, the LNP size is a significant factor in controlling the LNP biodistribution and LNP performance. Therefore, a precise LNP size control technique is indispensable for the LNP production process. Here, we report a protocol for size controlled LNP production using a microfluidic device, named iLiNP. siRNA loaded LNPs are also produced using the iLiNP device and evaluated by in vitro experiment. Representative results are shown for the LNP size, including siRNA-loaded LNPs, Z-potential, siRNA encapsulation efficiency, cytotoxicity, and target gene silencing activity.

Analysis of bacteria in the sewage wastewater treatment process is essential for process stabilization and upgrading. Although bacteria are currently being analyzed by molecular biology techniques targeting the 16S rRNA gene, there is a problem that they are time-consuming and labor-intensive. In this study, we developed a paper-based analytical chip by using two types of DNA molecules that specifically bind to bacterial 16S rRNA. We optimized the fabrication method of the detection probe and the paper-based analytical chip, and then detected synthetic DNA having a nucleotide sequence that hybridizes with the designed DNA molecules and bacterial 16S rRNA extracted from an activated sludge sample. We evaluated the amount of nucleic acids quantitatively by taking images of the detection line on the paper-based analytical chip with a smartphone and analyzing its brightness with an open-source image processing program, ImageJ. Our method was able to detect 85 nM of bacterial 16S rRNA concentration in the extract. Nucleic acids that did not hybridize with either of the designed DNA molecules were not detected, demonstrating high selectivity of our method.

Recent studies on nanopillar arrays, one type of nanofluidic device, have demonstrated various tools for bioanalysis. When carrying out nanopillar array-based separation, it is indispensable to obs...

Lithium carbonate is an effective medicine for the treatment of the bipolar disorder, but the concentration of lithium in the patient's blood must be frequently monitored because of its toxicity. To date, no colorimetric methods of lithium ion detection in whole blood without pretreatment have been reported. Here, we report a colorimetric paper-based device that allows point-of-care testing in one step. This device is composed of two paper-based elements linked to each other: a blood cell separation unit and a colorimetric detection unit. After a portion of whole blood has been placed on the end of the separation unit, plasma in the sample is automatically transported to the detection unit, which displays a diagnostic color. The key feature of this device is its simple, user-friendly operation. The limit of detection is 0.054 mM and the coefficient of variance is below 6.1%, which are comparable to those of conventional instruments using the same colorimetric reaction. Furthermore, we achieved high recovery (>90%) and reproducibility (

We report a colorimetric paper-based microfluidic device based on an enzyme inhibition assay that allows the on-site detection of nerve agents by sampling and wicking. The sample and reagents are automatically transported through the channel where an enzyme inhibition reaction is conducted, followed by an enzyme-substrate reaction and a color reaction. This device can detect 0.1 μg/mL of the nerve agent VX in a 2.5 μL drop and is nerve agent selective and robust against temperature, pH, and several liquids. We confirmed that sampling procedures (dilution and wiping) are applicable to this device. Furthermore, the fabrication procedure is easy, and the cost is at most a few tens of cents. Thus, the present device provides a practical method for the urgent detection of nerve agents in suspected chemical terrorism incidents.

Background Recurrent vasospastic angina sometimes occurs. Fresh thrombi have been known to arise without plaque rupture at coronary spasm sites due to blood flow stagnation and intimal erosion caused by vasospasms. The relationship between recurrence of vasospastic angina and thrombus formation remains unclear. Case summary A 67-year-old man presented with sudden chest pain at rest. Electrocardiography and coronary angiography indicated vasospastic angina. His chest pain persisted despite the administration of benidipine, isosorbide mononitrate, nicorandil, and nifedipine. Coronary angiography performed one month after initial presentation showed stenosis refractory to isosorbide administration. Optical coherence tomography revealed a healed plaque, and a stent was deployed. The patient remained symptom-free at 1-year follow-up. Discussion Prolonged coronary vasospasm with limited coronary blood flow could induce total occlusion of the coronary artery, and acute thrombus formation, which resulted in healed plaque erosion. When vasospastic angina cannot be controlled, rapidly progressive stenosis caused by healed plaque erosion could be its underlying cause and mechanism. This report indicates that antiplatelet therapy may be a preventive option for future recurrent vasospastic angina, especially in those caused by healed plaques.

Avian influenza virus (AIV) infection, caused by influenza virus type A, is an infectious, acute respiratory disease of birds related to influenza outbreaks worldwide. The highly pathogenic AIV subtype H5N1 has crossed species barriers to infect mammals, including humans, with fatal outcomes and has received attention as a potential pandemic threat. A rapid and timely detection in poultry is vitally important to prevent the virus spread. Despite their great sensitivity, conventional detection methods such as real-time reverse transcription-polymerase chain reaction and the agar gel precipitation test are time-consuming and labor-intensive and require special training. In this work, an immunowall device was evaluated as an easier and faster way for detecting AIV H5-hemagglutinin (AIV H5-HA). For detection, fluorescence-labeled or enzyme-labeled antibody was employed as a labeling antibody in a sandwich immunoassay. Both were shown in this paper to be easier and faster assays for detection compared with the conventional enzyme-linked immunosorbent assay (ELISA) kit. In addition, high selectivity was achieved for AIV H5-HA detection after the evaluation of other different HA virus subtypes. The limit of detection was 0.23 ng/mL for the enzyme-labeled antibody. This value was equivalent to that of the conventional ELISA kit but 8 times faster (31 min compared to 260 min). The detection range was 0.23-100 ng/mL. The immunowall device with the enzyme-labeled antibody offers a rapid, sensitive, selective, and simple immunoassay system for future H5 AIV real sample detection.

In this study, we developed an electrochemical sensor for ochratoxin A (OTA) by using an aptamer having a dithiol-based anchor, which exhibited higher stability on a gold electrode than a monothiol-based aptamer because of its two anchors. The sensor was also based on a signal-on scheme that produces a signal current resulting from structure-switching of the aptamer upon interaction with OTA. For simple fabrication of this sensor, the non-covalent interaction of methylene blue with the aptamer was also employed as an electrochemical indicator. In this study, the performance of the sensor, including the dissociation constant of the aptamer-OTA complex, was characterized. The proposed sensor exhibited high reproducibility and enough sensitivity to detect the minimum amount of OTA required for the analysis of real food samples with a limit of detection of 113 pM.

Fluorescence polarization (FP) is a one of the measurement techniques to study molecular interactions. We previously developed our own FP measurement system based on synchronization detection that uses a liquid crystal layer and an image sensor. The measurement cycle was fixed to 100 without any theoretical considerations, however, the influence of the synchrony and measurement cycles for FP values should be considered. In the present paper, we carried out an experimental and theoretical investigation into the influence of the synchrony between liquid crystal operation and image sampling for FP values of our system. When there was synchronization mismatch, the experimental FP values obtained using fluorescein ethylene glycol solution and the theoretical FP values changed according to the number of measurement cycles. Additionally, we measured the FP immunoassay for a physiologically active substance under synchronization mismatch. The synchronization mismatch influenced the measurement performance of the system, indicating that optimization of the number of image samplings was necessary to improve the measurement performance. For instance, the Mismatch 0.99 case, the measurement cycle should be 50 cycle judging from its dynamic range and R-square (R2). From the investigation, we obtained theoretical and experimental knowledge on FP response to facilitate further applications of our FP system.

We present an ultrasensitive immunoassay system for disease biomarkers utilizing the immuno-wall device and an enzymatic amplification reaction. The immuno-wall device consisted of 40 microchannels, each of which contained an antibody-modified wall-like structure along the longitudinal axis of the microchannel. The wall was fabricated with a water-soluble photopolymer containing streptavidin by photolithography, and biotinylated capture antibodies were immobilized on the sides through streptavidin-biotin interaction. For an assay, introducing the target biomarker and secondary and labeled antibodies produced a sandwich complex anchored on the sides of the wall. A conventional immuno-wall device uses a fluorescence-labeled antibody as a labeling antibody. To achieve an ultrasensitive detection of a trace biomarker, we used an enzyme label and amplified the signal with the enzymatic reaction with a fluorogenic substrate in the microchannel. The highest signal/background ratio was obtained by using alkaline phosphatase-labeled antibody and 9H-(1,3-dichloro-9,9-dimethylacridin-2-one-7-yl) phosphate. To evaluate the device performance, we detected human C-reactive protein (CRP) as a model biomarker. The detection limit (LOD) of CRP in phosphate-buffered saline was 2.5 pg mL-1 with a sample volume of 0.25 μL. This LOD was approximately 3 orders of magnitude lower than that obtained with fluorescent-dye (DyLight 650)-labeled antibody. In addition, the present device provided a wide detection range of 0.0025-10 ng mL-1 for CRP. We successfully developed an ultrasensitive immunoassay system with simple operation and only a small sample volume.

High-throughput fluorescence polarization immunoassays (FPIAs) for mycotoxin were conducted using a portable FP analyzer with a microdevice. Simultaneous FPIA measurements for 8 different deoxynivalenol (DON) concentrations in 12 chambers (total of 96 samples) and high-throughput FPIA measurements for single DON concentrations in more than 500 chambers were conducted. The results indicated that simultaneous FPIAs for 96 independent samples and for 500 samples were possible by FP imaging. The FP analyzer has a size of 65 cm (W 35 cm × D 15 cm × H 15 cm) and costs less than $5000. The sample volume was 1 nL. Furthermore, it is expected that sample reaction and FP detection can be automatically conducted with the analyzer by changing the microdevice and the software. Its features such as low cost and portability will contribute to on-site measurement and point-of-care testing. Additionally, the high-throughput feature will contribute to the study of molecular interactions based on FP measurements.

Sub 100 nm-sized lipid nanoparticles (LNPs) have been widely used in drug delivery systems (DDSs). The size of the LNPs is an important parameter for the DDS performance, such as biodistribution and gene silencing using siRNAs. However, the LNPs prepared by the conventional preparation method show a wide size distribution. To improve the LNP size distribution, we developed a microfluidic device, named the iLiNP™ device, in a previous study. This device could produce LNPs in the size range of 20 to 150 nm, but the size distribution of the large-sized LNPs needs to be further improved. From the viewpoint of the LNP formation process, a homogeneous and slow rate dilution of ethanol plays an important role in improving the large-size LNP size distribution. In this study, we developed a three-dimensional, symmetrically assembled microfluidic device named the 3D-iLiNP device with the aim of precise size control of large-sized LNPs. We designed the 3D-iLiNP device using a computational fluid dynamics simulation and demonstrated that the 3D-iLiNP device can improve the LNP size distribution. The gene silencing activity of four kinds of siRNA-loaded LNPs was investigated via in vitro and in vivo experiments to elucidate the effect of the LNP size distribution. The results revealed that the LNPs with a size between 90 and 120 nm showed higher gene silencing activity than those with other sizes. The 3D-iLiNP device is expected to improve DDS performance by precisely controlling the size of LNPs.

Microfluidic methodologies for preparation of lipid nanoparticles (LNPs) based on an organic solvent injection method enable precise size control of the LNPs. After preparation of LNPs, the organic solvent injection method needs some post-treatments, such as overnight dialysis or direct dilution with a buffer solution. LNP production using the microfluidic-based organic solvent injection method is dominated by kinetics rather than thermodynamics. Kinetics of ethanol removal from the inner and outer membranes of LNPs could induce a structural change in the membrane that could lead to fusion of LNPs. However, the effects of microfluidic post-treatment on the final size of LNPs have not been sufficiently understood. Herein, we investigated the effect of the post-treatment processes on the final product size of LNPs in detail. A simple baffle device and a model lipid system composed of a neutral phospholipid (1-palmitoyl-2-oleoyl

This study aimed to develop an electrochemical system for measuring blood ATP and lactate levels in a single format. The ratio of ATP to lactate levels was previously reported to provide an alternative illness severity score. Although severity evaluation is crucial to treat patients with acute disease admitted to intensive care units, no sensors are currently available to simply and rapidly measure ATP and lactate levels using the same detection method. Therefore, we constructed an integrated sensing system for ATP and lactate using enzymatic reactions and two sets of electrodes integrated into a chip connected to a single potentiostat operated by a microcontroller. The enzymatic system involves adenylate kinase, pyruvate kinase, and pyruvate oxidase for ATP, and lactate oxidase for lactate, both of which produce hydrogen peroxide. Multiplex enzyme-based reactions were designed to minimize the corresponding operations significantly without enzyme immobilization onto the electrodes. The system was robust in the presence of potentially interfering blood components, such as ascorbate, pyruvate, ADP, urate, and potassium ions. The ATP and lactate levels in the blood were successfully measured using the new sensor with good recoveries. The analytical results of blood samples obtained using our sensor were in good agreement with those using conventional methods. Integrating electrode-based analysis and a microcontroller-based system saved further operations, enabling the straightforward measurement of ATP and lactate levels within 5 min. The proposed sensor may serve as a useful tool in the management of serious infectious diseases.