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13
result(s) for
"Kasama, Toshihiro"
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Development of cellulosic material-based microchannel device capable of fluorescence immunoassay of microsamples
2022
Microfluidic immunoassay devices are a promising technology that can quickly detect biomarkers with high sensitivity. Recently, many studies implementing this technology on paper substrates have been proposed for improving cost and user-friendliness. However, these studies have identified problems with the large volume of sample required, low sensitivity, and a lack of quantitative accuracy and precision. In this paper, we report a novel structure implemented as a cellulosic material-based microchannel device capable of quantitative immunoassay using small sample volumes. We fabricated microfluidic channels between a transparent cellophane film and water-resistant paper to facilitate loading of small-volume samples and reagents, with a 40-μm-wide immunoreaction matrix constructed in the center of the microchannel using highly precise photolithography. A fluorescence sandwich immunoassay for C-reactive protein (CRP) was successfully implemented that required only a 1-μL sample volume and a 20-min reaction time. We confirmed that the limit of detection of the device was 10–20 ng/mL with a coefficient of variation under 5.6%, which is a performance level comparable to conventional plastic-based human CRP enzyme-linked immunosorbent assay (ELISA) kits. We expect that such devices will lead to the elimination of large amounts of medical waste from the use of ubiquitous diagnostics, a result that is consistent with environmental sustainability goals.
Journal Article
A Mediated Enzymatic Electrochemical Sensor Using Paper-Based Laser-Induced Graphene
2022
Laser-induced graphene (LIG) has been applied in many different sensing devices, from mechanical sensors to biochemical sensors. In particular, LIG fabricated on paper (PaperLIG) shows great promise for preparing cheap, flexible, and disposable biosensors. Distinct from the fabrication of LIG on polyimide, a two-step process is used for the fabrication of PaperLIG. In this study, firstly, a highly conductive PaperLIG is fabricated. Further characterization of PaperLIG confirmed that it was suitable for developing biosensors. Subsequently, the PaperLIG was used to construct a biosensor by immobilizing glucose oxidase, aminoferrocene, and Nafion on the surface. The developed glucose biosensor could be operated at a low applied potential (−90 mV) for amperometric measurements. The as-prepared biosensor demonstrated a limit of detection of (50–75 µM) and a linear range from 100 µM to 3 mM. The influence of the concentration of the Nafion casting solution on the performance of the developed biosensor was also investigated. Potential interfering species in saliva did not have a noticeable effect on the detection of glucose. Based on the experimental results, the simple-to-prepare PaperLIG-based saliva glucose biosensor shows great promise for application in future diabetes management.
Journal Article
Evaluation of lung epithelial lining fluid concentrations of lascufloxacin against Streptococcus pneumoniae in a hollow-fiber infection model
by
Hayashi, Yukitaka
,
Hamada, Masakaze
,
Matsumoto, Kazuaki
in
631/154/1435/2417
,
631/326/22/1290
,
Aluminum
2025
Lascufloxacin (LSFX) achieves concentrations in epithelial lining fluid (ELF) that are more than 15 times higher than those in the bloodstream, making it a promising candidate for respiratory and otorhinolaryngological infections. These concentrations were replicated using the Hollow-Fiber Infection Model and demonstrated bactericidal efficacy of LSFX against levofloxacin-sensitive and -resistant
Streptococcus pneumoniae
. The study confirms that LSFX’s elevated concentration in ELF plays a significant role in its bactericidal activity.
Journal Article
A needle-type micro-sampling device for collecting nanoliter sap sample from plants
2021
In plant research, measuring the physiological parameters of plants is vital for understanding the behavior and response of plants to changes in the external environment. Plant sap analysis provides an approach for elucidating the physiological condition of plants. However, to facilitate accurate sap analysis, a sampling device capable of collecting sap samples from plants is required. In this paper, a minimally invasive, needle-type micro-sampling device capable of collecting nanoliter (~ 91 nL) quantities of sap from plants is described. The developed micro-sampling system showed great reproducibility (3%) in experiments designed to assess sampling performance. As a proof of concept, sap samples were collected continuously from target plants with the micro-sampling system, and the dynamic changes in potassium ions, plant hormones and sugar levels inside plants were analyzed. The results demonstrated the feasibility of the micro-sampling device and its potential for developing a measurement system for plant research in the future.
Journal Article
Democratizing Microreactor Technology for Accelerated Discoveries in Chemistry and Materials Research
by
Chikako Sano
,
Koji Masuda
,
Tomomi Sato
in
accelerated discovery
,
Accelerators
,
advanced integrated microreactor development platform
2024
Microreactor technologies have emerged as versatile platforms with the potential to revolutionize chemistry and materials research, offering sustainable solutions to global challenges in environmental and health domains. This survey paper provides an in-depth review of recent advancements in microreactor technologies, focusing on their role in facilitating accelerated discoveries in chemistry and materials. Specifically, we examine the convergence of microfluidics with machine intelligence and automation, enabling the exploitation of the cyber-physical environment as a highly integrated experimentation platform for rapid scientific discovery and process development. We investigate the applicability and limitations of microreactor-enabled discovery accelerators in various chemistry and materials contexts. Despite their tremendous potential, the integration of machine intelligence and automation into microreactor-based experiments presents challenges in establishing fully integrated, automated, and intelligent systems. These challenges can hinder the broader adoption of microreactor technologies within the research community. To address this, we review emerging technologies that can help lower barriers and facilitate the implementation of microreactor-enabled discovery accelerators. Lastly, we provide our perspective on future research directions for democratizing microreactor technologies, with the aim of accelerating scientific discoveries and promoting widespread adoption of these transformative platforms.
Journal Article
Hollow Fiber Microreactor Combined with Digital Twin to Optimize the Antimicrobial Evaluation Process
by
Shinohara, Marie
,
Noda, Kazuhiro
,
Hamada, Masakaze
in
3-D printers
,
Analysis
,
Antimicrobial agents
2024
In order to reproduce pharmacokinetics (PK) profiles seen in vivo, the Hollow Fiber Infection Model (HFIM) is a useful in vitro module in the evaluation of antimicrobial resistance. In order to reduce the consumption of culture medium and drugs, we developed a hollow fiber microreactor applicable to the HFIM by integrating the HFIM function. Next, we constructed a novel control method by using the “digital twin” of the microreactor to achieve precise concentration control. By integrating functions of the HFIM, the extra-capillary space volume was reduced to less than 1/10 of conventional HFIM. The control method with the digital twin can keep drug concentration in the extra-capillary space within an error of 10% under simulated drug destruction. The control method with the digital twin can also stabilize the drug concentration both in the intra-capillary space and the extra-capillary space within 15 min.
Journal Article
Development of an immuno-wall device for the rapid and sensitive detection of EGFR mutations in tumor tissues resected from lung cancer patients
by
Sato, Mitsuo
,
Kasama, Toshihiro
,
Kawabe, Nozomi
in
Antibodies
,
Assaying
,
Biology and Life Sciences
2020
Detecting molecular targets in specimens from patients with lung cancer is essential for targeted therapy. Recently, we developed a highly sensitive, rapid-detection device (an immuno-wall device) that utilizes photoreactive polyvinyl alcohol immobilized with antibodies against a target protein via a streptavidin–biotin interaction. To evaluate its performance, we assayed epidermal growth factor receptor ( EGFR ) mutations, such as E746_A750 deletion in exon 19 or L858R substitution in exon 21, both of which are common in non-small cell lung cancer and important predictors of the treatment efficacy of EGFR tyrosine kinase inhibitors. The results showed that in 20-min assays, the devices detected as few as 1% (E746_A750 deletion) and 0.1% (L858R substitution) of mutant cells. Subsequent evaluation of detection of the mutations in surgically resected lung cancer specimens from patients with or without EGFR mutations and previously diagnosed using commercially available, clinically approved genotyping assays revealed diagnostic sensitivities of the immuno-wall device for E746_A750 deletion and L858R substitution of 85.7% and 87.5%, respectively, with specificities of 100% for both mutations. These results suggest that the immuno-wall device represents a good candidate next-generation diagnostic tool, especially for screening of EGFR mutations.
Journal Article
Rapid and highly sensitive immunoassay using an ultra-thin immuno-wall microfluidic device with a sequential fluorescence signal increment method
by
Miyake, Ryo
,
Zhou, Xiang
,
Kasama, Toshihiro
in
Analytical Chemistry
,
Antibodies
,
Antibodies, Immobilized - chemistry
2025
We present a rapid and highly sensitive immunoassay platform based on an ultra-thin immuno-wall microfluidic device with an easy-to-perform sequential fluorescence signal increment method. The ultra-thin immuno-wall was fabricated using a special type of water-soluble photopolymer mixed with streptavidin via photolithography. During photolithography, the photopolymer formed a three-dimensional cross-linked structure, and streptavidin was immobilized in the cross-linked structure based on the click chemistry reaction. The immobilized streptavidin was used to immobilize biotin-conjugated antibodies on the cross-linked structure to capture biomarkers, forming immune complexes on the surface, known as an “immuno-wall.” A sequential fluorescence signal increment method utilizes two different fluorescence-labeled antibodies with high affinity that were incubated several cycles in the immuno-wall to enhance the fluorescence signal. Moreover, an ultra-thin immuno-wall was developed to reduce the nonspecific binding and increase the signal-to-noise ratio. To evaluate the performance of this immunoassay platform, the spike protein from the SARS-CoV-2 virus was selected as the target biomarker. This immunoassay platform exhibited a limit of detection of 0.01 ng/mL, and the detection time was 30 min, which is comparable to rapid antigen tests. This immunoassay platform demonstrates significant potential for early-phase disease diagnosis.
Journal Article
Microchip Immunoassays for Monitoring Renal Function: Rapid, Low-Cost, and Highly Sensitive Quantification of Urinary Biomarkers of Diabetic Nephropathy
2021
This study developed low-cost and highly sensitive immunoassay devices possessing the ability to rapidly analyze urine samples. Further, they can quantitatively detect three biomarkers indicating renal injury: monocyte chemotactic protein 1 (MCP-1), angiotensinogen (AGT), and liver-type fatty acid binding protein (L-FABP). The devices were used to successfully estimate the concentrations of the three biomarkers in urine samples within 2 min; the results were consistent with those obtained via conventional enzyme-linked immunosorbent assay (ELISA), which requires several hours. In addition, the estimated detection limits for the three biomarkers were comparable to those of commercially available ELISA kits. Thus, the proposed and fabricated devices facilitate high-precision and frequent monitoring of renal function.
Journal Article
An immuno-wall microdevice exhibits rapid and sensitive detection of IDH1-R132H mutation specific to grade II and III gliomas
by
Kato, Yukinari
,
Suzuki, Hidenori
,
Ranjit, Melissa
in
30 Bio-inspired and biomedical materials
,
404 Materials informatics / Genomics
,
Antibodies
2016
World Health Organization grade II and III gliomas most frequently occur in the central nervous system (CNS) in adults. Gliomas are not circumscribed; tumor edges are irregular and consist of tumor cells, normal brain tissue, and hyperplastic reactive glial cells. Therefore, the tumors are not fully resectable, resulting in recurrence, malignant progression, and eventual death. Approximately 69-80% of grade II and III gliomas harbor mutations in the isocitrate dehydrogenase 1 gene (IDH1), of which 83-90% are found to be the IDH1-R132H mutation. Detection of the IDH1-R132H mutation should help in the differential diagnosis of grade II and III gliomas from other types of CNS tumors and help determine the boundary between the tumor and normal brain tissue. In this study, we established a highly sensitive antibody-based device, referred to as the immuno-wall, to detect the IDH1-R132H mutation in gliomas. The immuno-wall causes an immunoreaction in microchannels fabricated using a photo-polymerizing polymer. This microdevice enables the analysis of the IDH1 status with a small sample within 15 min with substantially high sensitivity. Our results suggested that 10% content of the IDH1-R132H mutation in a sample of 0.33 μl volume, with 500 ng protein, or from 500 cells is theoretically sufficient for the analysis. The immuno-wall device will enable the rapid and highly sensitive detection of the IDH1-R132H mutation in routine clinical practice.
Journal Article