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Miniaturized electrochemiluminescent biochip prepared on gold nanoparticles-loaded mesoporous silica film for visual detection of hydrogen peroxide released from living cells
摘要： Au nanoparticles (NPs) has been widely used for the detection of intracellular H2O2 to enhance the electron transfer process. But AuNPs are easy to aggregate in the live cells environment. Herein we report a rapid, reliable and low-cost electrochemiluminescent (ECL) biochip integrated by AuNPs-loaded mesoporous silica film (MSF) to detect H2O2 released by macrophage cells. The MSF was employed as a template to load AuNPs within the nanochannels to avoid aggregation. H2O2 could be catalyzed by AuNPs to promote the ECL reaction of luminol molecules in solution. The ECL intensity was significantly enhanced, and the peak potential was negatively shifted by 400 mV due to the excellent electrocatalytic ability of AuNPs. The integrated biochip demonstrated good reproducibility, with a wide linear range of 0.1–200 μM and an LOD of 25.3 nM. The reliability was evaluated by applying for the assessment of antioxidant activity of resveratrol using RAW 264.7 macrophage model. The AuNPs-loaded MSF integrated biochip can be easily adapted to the development of improved devices in biosensing, lab-on-a-chip, and nanofluidic systems.
关键词： Gold nanoparticles,Biochip array,Electrochemiluminescence,Reactive oxygen species,Mesoporous silica
更新于2025-09-23 15:23:52
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Validation de l’immunofluorescence indirecte en mosa?que versus technique de référence IFI et Elisa dans les maladies bulleuses auto-immunes
摘要： L’immunofluorescence indirecte (IFI) permet de mieux typer la dermatose bulleuse auto-immune (DBAI). L’étude décrit une technique d’IFI en mosa?que. Il s’agit d’une IFI sur mosa?que de six substrats (BIOCHIP Mosa?csTM) : ?sophage de primate, peau humaine traitée par une solution de NaCl, cellules exprimant les desmogléines 1 et 3, la BP230 et la BP180. Chaque substrat est dans une biopuce pour une analyse simultanée. Elle consiste à mettre en contact le sérum des patients avec les substrats. Si une réaction positive est obtenue, les autoanticorps de classes IgA, IgG et IgM présents se fixent au déterminant antigénique de la protéine visée et ensuite le complexe antigène—anticorps est révélé par un anticorps (IgG) marqué au fluorochrome, et est visualisé à l’aide d’un microscope à fluorescence conventionnel. Il s’agit d’une étude diagnostique monocentrique réalisée en 2015 avec des sérums de 2012 à 2015. Quatre-vingt sérums au total testés sur 67 patients (47 pemphigo?des bulleuses [PB] et 2 pemphigo?des cicatricielles, 16 pemphigus vulgaires, 9 pemphigus superficiels, 2 épidermolyses bulleuses acquises). Quatre témoins négatifs ont été utilisés. Chaque élément de la mosa?que est comparé avec la technique de référence (IFI sur peau humaines provenant de reliquats chirurgicaux avec peau splitée). Soixante-dix-neuf sérums ont été analysés, pour 40 sérums, il est réalisé une triple lecture et pour 39 une double lecture. En utilisant l’IFI en mosa?que par rapport à la technique de référence, les sensibilités toutes maladies confondues de la desmogléine 1, desmogléine 3, BP180 NC16A étaient respectivement de : 100 %, 92 %, 85 %. L’antigène BP 230 était reconnu dans 59 % des PB. Pour tous les substrats sauf la peau séparée, la spécificité variait de 84 à 100 %. Pour la peau séparée, elle était de 60 %. Parmi les 40 sérums ayant eu une triple lecture pour 31 patients les résultats étaient identiques pour les 3 lecteurs (78 %). Parmi les 39 sérums ayant eu une double lecture pour 35 les résultats étaient identiques pour les deux lectures (90 %). Les résultats de notre étude sont en accord avec ceux rapportés dans les études retrouvées dans la littérature. L’IFI en mosa?que permet d’avoir un spectre antigénique complet disponible en une seule analyse (un seul test qui permet de différencier les principales DBAI) ; de différencier le pemphigus superficiel et vulgaire. Elle permet une interprétation simple sans spectrophotométrie, les résultats sont rapides en une journée. Elle est facilement reproductible. L’IFI en mosa?que a une bonne spécificité et sensibilité pour le diagnostic des DBAI. C’est une méthode standardisée, plus rapide, pour faciliter le diagnostic de ces maladies. Cette technique s’inscrit dans le cadre de la certification des laboratoires dans la mesure ou l’utilisation de substrats humains ou animaux n’est plus autorisée.
关键词： Immunofluorescence indirecte,BIOCHIP technologie,auto-immune,Dermatose bulleuse
更新于2025-09-23 15:22:29
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A versatile fluorometric in-situ hybridization method for the quantitation of hairpin conformations in DNA self-assembled monolayers
摘要： As the performance of hairpin DNA (hpDNA)-based biosensors is highly dependent on the yield of stem-loop (hairpin) conformations, we report herein a versatile fluorometric in-situ hybridization protocol for examining hpDNA self-assembled monolayers (SAMs) on popularly used biochip substrates. Specifically, the ratio of fluorescence (FL) intensities of hpDNA SAMs (in an array format) before and after hybridization was adapted as the key parameter for performing such a determination. Upon confirming the existence of mixed and tunable DNA conformations in binary deposition solutions and the efficient hybridization of the hairpin strands with target DNA via gel electrophoresis assays, we have tested the fluorometric protocol for determining the coverages of hpDNA in hpDNA/ssDNA SAMs prepared on gold; its accuracy was validated by Exonuclease I (Exo I)-assisted electrochemical quantitation. To further confirm its versatility, this FL protocol was adapted for quantifying hairpin conformations formed on glass and polycarbonate (PC) substrates. The molar ratios of surface-tethered hairpin conformations on the three different substrates were all found to be proportional to but less than that in the binary deposition solutions, and dependent on the substrate morphology. The findings reported herein are beneficial to the construction of highly efficient DNA hairpin-based sensing surfaces, which essentially facilitates the creation of hpDNA-based biosensors with optimal detection performance.
关键词： In-situ hybridization,conformation evaluation,DNA biosensor/biochip,fluorescence microarray,Hairpin DNA
更新于2025-09-23 15:19:57
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A fluorescent plasmonic biochip assay for multiplex screening of diagnostic serum antibody targets in human Lyme disease
摘要： Lyme disease (LD) diagnosis using the current two-tier algorithm is constrained by low sensitivity for early-stage infection and ambiguity in determining treatment response. We recently developed a protein microarray biochip that measures diagnostic serum antibody targets using grating-coupled fluorescent plasmonics (GC-FP) technology. This strategy requires microliters of blood serum to enable multiplexed biomarker screening on a compact surface and generates quantitative results that can be further processed for diagnostic scoring. The GC-FP biochip was used to detect serum antibodies in patients with active and convalescent LD, as well as various negative controls. We hypothesized that the quantitative, high-sensitivity attributes of the GC-FP approach permit: 1) screening of antibody targets predictive for LD status, and 2) development a diagnostic algorithm that is more sensitive, specific, and informative than the standard ELISA and Western blot assays. Notably, our findings led to a diagnostic algorithm that may be more sensitive than the current standard for detecting early LD, while maintaining 100% specificity. We further show that analysis of relative antibody levels to predict disease status, such as in acute and convalescent stages of infection, is possible with a highly sensitive and quantitative platform like GC-FP. The results from this study add to the urgent conversation regarding better diagnostic strategies and more effective treatment for patients affected by tick-borne disease.
关键词： Lyme disease,grating-coupled fluorescent plasmonics (GC-FP) technology,diagnostic serum antibody targets,multiplex screening,fluorescent plasmonic biochip assay
更新于2025-09-19 17:13:59
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[IEEE 2018 IEEE 27th Asian Test Symposium (ATS) - Hefei, China (2018.10.15-2018.10.18)] 2018 IEEE 27th Asian Test Symposium (ATS) - Test Diagnosis of Digital Microfluidic Biochips Using Image Segmentation
摘要： Digital micro?uidic biochip has been developed as a promising alternative to the traditional approach of benchtop laboratory tests. Dependability is an important biochemical attribute for micro?uidic biochips that are used for safety-critical applications, such as point-of-care health assessment, air-quality monitoring, and food-safety testing. Therefore, the robust of?ine and online test techniques are required after manufacturing and during bioassay operations. In this work, we are presenting an image segmentation based testing methodology to detect the catastrophic faults and to locate the faulty cells. The design-for diagnosability scheme is proposed, and it is shown that faults can be located and tolerated by providing alternative paths in biochips. Moreover this testing method also facilitates the testing of a biochip with other bioassay operations running concurrently.
关键词： fault models,CCD,Biochip,electrowetting,image segmentation,test time
更新于2025-09-04 15:30:14