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1.
J Immunoassay Immunochem ; 44(1): 76-89, 2023 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-36318041

RESUMO

Membrane proteins are difficult to be extracted and to be coated on the substrate of the immunoassay reaction chamber because of their hydrophobicity. Traditional method to prepare membrane protein sample requires many steps of protein extraction and purification that may lead to protein structure deformation and protein dysfunction. This work proposes a simple technique to prepare and immobilize the membrane protein suspended in an unprocessed crude cell lysate sample. Membrane fractions in crude cell lysate were incorporated with the large unilamellar vesicle (LUV) that was mainly composed of POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) before coating in the polystyrene plate by passive adsorption technique. Immunofluorescence staining and the Enzyme-Linked Immunosorbent Assay (ELISA) examination of a strictly conformation-dependent integral membrane protein, Myelin Oligodendrocyte Glycoprotein (MOG), demonstrate that LUV incorporated cell lysate sample obviously promotes MOG protein immobilization in the microplate well. With LUV incorporation, the dose-response curve of the MOG transfected cell lysate coating plate can be 2-9 times differentiated from that of the untransfected cell lysate coating plate. The LUV incorporated MOG transfected cell lysate can be efficiently coated in the microplate without carbonate/bicarbonate coating buffer assistance.


Assuntos
Proteínas de Membrana , Imunoensaio/métodos , Ensaio de Imunoadsorção Enzimática/métodos
2.
Micromachines (Basel) ; 13(12)2022 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-36557545

RESUMO

The optimal groove design of a MEMS piezoresistive pressure sensor for ultra-low pressure measurement is proposed in this work. Two designs of the local groove and one design of the annular groove are investigated. The sensitivity and linearity of the sensor are investigated due to the variations of two dimensionless geometric parameters of these grooves. The finite element method is used to determine the stress and deflection of the diaphragm in order to find the sensor performances. The sensor performances can be enhanced by creating the annular or local groove on the diaphragm with the optimal dimensionless groove depth and length. In contrast, the performances are diminished when the local groove is created on the beam at the piezoresistor. The sensitivity can be increased by increasing the dimensionless groove length and depth. However, to maintain low nonlinearity error, the annular and local grooves should be created on the top of the diaphragm. With the optimal designs of annular and local grooves, the net volume of the annular groove is four times greater than that of the local groove. Finally, the functional forms of the stress and deflection of the diaphragm are constructed for both annular and local groove cases.

3.
Int J Mol Sci ; 23(12)2022 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-35742849

RESUMO

In addition to their use as an additive to improve physical properties of solvent polymeric membranes, plasticizers have a considerable impact on the specificity and sensitivity of membrane-modified electrochemical sensors. In this work, we aim at the hybridization of two different plasticizers using the electropolymerization technique in the development of a cadmium(II)-selective electrochemical sensor based on screen-printed gold electrode along with cyclic voltammetric measurement. At this point, we first screen for the primary plasticizer yielding the highest signal using cyclic voltammetry followed by pairing it with the secondary plasticizers giving rise to the most sensitive current response. The results show that the hybridization of DOS and TOTM with 3:1 weight ratio (~137.7-µm-thick membrane) renders a signal that is >26% higher than that from the sensor plasticized by DOS per se in water. The solution of 0.1 mM hydrochloric acid (pH 4) is the optimal supporting electrolyte. In addition, hybrid plasticizers have adequate redox capacity to induce cadmium(II) transfer from bulk solution to the membrane/water interfaces. Conversion of voltammetric signals to semi-integral currents results in linearity with cadmium(II) concentration, indicating the irreversible cadmium(II) transfer to the membrane. The DOS:TOTM hybrid sensor also exhibits high sensitivity, with a limit of detection (LOD) and limit of quantitation (LOQ) of 95 ppb and 288 ppb, respectively, as well as greater specificity towards cadmium(II) than that obtained from the single plasticizer sensor. Furthermore, recovery rates of spiked cadmium(II) in water samples were higher than 97% using the hybrid plasticizer sensor. Unprecedentedly, our work reports that the hybridization of plasticizers serves as ion-to-electron transducer that can improve the sensor performance in cadmium(II) detection.


Assuntos
Cádmio , Plastificantes , Cádmio/química , Eletrodos , Ouro , Água
4.
Front Vet Sci ; 9: 1048131, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36686171

RESUMO

The rectangular filtering microfluidic chip was invented using microfluidics device fabrication technology and can separate living microfilariae from blood samples without a syringe pump. The diagnostic results are highly effective. The device is based on the principle of separating millions of blood cells from microfilariae using a rectangular filter structure. It disperses fluid evenly into the flow-passage channel, and its rectangular filter structure is the key to success in reducing the pressure and separating blood cells from microfilariae effectively. The flow rate and blood cell concentration were optimized in our study. The chip is intended to be a point-of-care device that can reduce the use of superfluous instrumentation in the field. The technology is designed to be rapid, accurate, and easy-to-use for all users, especially those in remote areas.

5.
Micromachines (Basel) ; 14(1)2022 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-36677084

RESUMO

Dynamic gut-on-a-chip platform allows better recreation of the intestinal environment in vitro compared to the traditional static cell culture. However, the underlying mechanism is still not fully discovered. In this study, the shear stress behavior in a gut-on-a-chip device with porous membrane subjected to peristalsis motion is numerically investigated using CFD simulation for three different pore sizes and two pattern layouts. The results reveal that, in the stationary microchannel, the average shear stress on the porous membrane is approximately 15% greater than that of the flat membrane, regardless of the pore size. However, when subjected to cyclic deformation, the porous membrane with smaller pore size experiences stronger variation of shear stress which is ±5.61%, ±10.12% and ±34.45% from its average for the pore diameters of 10 µm, 5 µm and 1 µm, respectively. The shear stress distribution is more consistent in case of the staggered pattern layout while the in-line pattern layout allows for a 32% wider range of shear stress at the identical pore size during a cyclic deformation. These changes in the shear stress caused by peristalsis motion, porous size and membrane pattern could be the key factors that promote cell differentiation in the deforming gut-on-a-chip model.

6.
Diagnostics (Basel) ; 11(10)2021 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-34679553

RESUMO

Lymphatic filariasis (LF) is a neglected major tropical disease that is a leading cause of permanent and long-term disability worldwide. Significant progress made by the Global Programme to Eliminate Lymphatic Filariasis (GPELF) has led to a substantial decrease in the levels of infection. In this limitation, DNA detection of lymphatic filariae could be useful due to it capable of detecting low level of the parasites. In the present study, we developed a diagnostic assay that combines a miniPCR with a duplex lateral flow dipstick (DLFD). The PCR primers were designed based on the HhaI and SspI repetitive noncoding DNA sequences of Brugia malayi and Wuchereria bancrofti, respectively. The limits of detection and crossreactivity of the assay were evaluated. In addition, blood samples were provided by Thais living in a brugian filariasis endemic area. The miniPCR-DLFD assay exhibited a detection limit of 2 and 4 mf per milliliter (mL) of blood for B. malayi as well as W. bancrofti, respectively, and crossamplification was not observed with 11 other parasites. The result obtained from the present study was in accordance with the thick blood smear staining for the known cases. Thus, a miniPCR-DLFD is an alternative tool for the diagnosis of LF in point-of-collection settings with a modest cost (~USD 5) per sample.

7.
Nanomaterials (Basel) ; 11(2)2021 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-33572813

RESUMO

Bio-inspired surfaces with superamphiphobic properties are well known as effective candidates for antifouling technology. However, the limitation of large-area mastering, patterning and pattern collapsing upon physical contact are the bottleneck for practical utilization in marine and medical applications. In this study, a roll-to-plate nanoimprint lithography (R2P NIL) process using Morphotonics' automated Portis NIL600 tool was used to replicate high aspect ratio (5.0) micro-structures via reusable intermediate flexible stamps that were fabricated from silicon master molds. Two types of Morphotonics' in-house UV-curable resins were used to replicate a micro-pillar (PIL) and circular rings with eight stripe supporters (C-RESS) micro-structure onto polycarbonate (PC) and polyethylene terephthalate (PET) foil substrates. The pattern quality and surface wettability was compared to a conventional polydimethylsiloxane (PDMS) soft lithography process. It was found that the heights of the R2P NIL replicated PIL and C-RESS patterns deviated less than 6% and 5% from the pattern design, respectively. Moreover, the surface wettability of the imprinted PIL and C-RESS patterns was found to be superhydro- and oleophobic and hydro- and oleophobic, respectively, with good robustness for the C-RESS micro-structure. Therefore, the R2P NIL process is expected to be a promising method to fabricate robust C-RESS micro-structures for large-scale anti-biofouling application.

8.
RSC Adv ; 11(56): 35653-35662, 2021 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-35493190

RESUMO

The objective of this paper is to propose a surface modification method for preparing PDMS microfluidic devices with partially hydrophilic-hydrophobic surfaces for generating double emulsion droplets. The device is designed to be easy to use without any complicated preparation process and also to achieve high droplet encapsulation efficiency compared to conventional devices. The key component of this preparation process is the permanent chemical coating for which the Pluronic surfactant is added into the bulk PDMS. The addition of Pluronic surfactant can modify the surface property of PDMS from a fully hydrophobic surface to a partially hydrophilic-hydrophobic surface whose property can be either hydrophilic or hydrophobic depending on the air- or water-treatment condition. In order to control the surface wettability, this microfluidic device with the partially hydrophilic-hydrophobic surface undergoes water treatment by injecting deionized water into the specific microchannels where their surface property changes to hydrophilic. This microfluidic device is tested by generating monodisperse water-in-oil-in-water (w/o/w) double emulsion micro-droplets for which the maximum droplet encapsulation efficiency of 92.4% is achieved with the average outer and inner diameters of 75.0 and 57.7 µm, respectively.

9.
Talanta ; 207: 120305, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31594628

RESUMO

The 3-aminopropyltriethoxysilane (APTES) is a common method for biomolecule immobilization on silicon and silicon derivatives such as silicon nitride (Si3N4). However, there are many parameters which impact the efficiency of APTES modification such as APTES concentration and reaction time. Thus, various APTES concentrations (0.1%, 0.5%, 1%, 2%, 5%, and 10%) under different reaction times (15, 30, 60 and 120 min) were compared to achieve the optimal APTES modification condition which produced a thin and stable APTES layer on Si3N4 surface. The modified surfaces were characterized by contact angle (CA) measurement, Fourier transform infrared (FTIR) spectroscopy and spectroscopic ellipsometry to determine the wetting property, chemical bonding composition and surface thickness, respectively. In addition, biotin was used as a model to determine the effectiveness of APTES modification condition by coupling with glutaraldehyde (GA). The Alexa Flour 488 conjugated streptavidin was performed to visualize the presence of biotin using fluorescence microscopy due to the specifically binding between biotin and streptavidin. The atomic force microscopy (AFM) was utilized to determine the surface topology which was an indicator to demonstrate the agglomeration of APTES molecule. Moreover, ion sensitive field effect transistor (ISFET) was employed as a biosensor model to demonstrate the effect between surface thickness and sensitivity of biosensor. The results show that the APTES thickness is directly correlated to the APTES concentration and reaction time. Since the importance parameter for ISFET measurement is the distance between biomolecule and sensing membrane of ISFET, the thicker APTES layer negatively impacts the sensitivity of ISFET based biosensor because of the ion shielding effect. Therefore, these results would be valuable information for development of Si3N4 biosensor, especially ISFET based biosensor.


Assuntos
Técnicas Biossensoriais/métodos , Propilaminas/química , Silanos/química , Compostos de Silício/química , Glutaral/química , Cinética , Propriedades de Superfície , Transistores Eletrônicos
10.
Micromachines (Basel) ; 10(12)2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31805714

RESUMO

Cellular heterogeneity is a major hindrance, leading to the misunderstanding of dynamic cell biology. However, single cell analysis (SCA) has been used as a practical means to overcome this drawback. Many contemporary methodologies are available for single cell analysis; among these, microfluidics is the most attractive and effective technology, due to its advantages of low-volume specimen consumption, label-free evaluation, and real-time monitoring, among others. In this paper, a conceptual application for microfluidic single cell analysis for veterinary research is presented. A microfluidic device is fabricated with an elastomer substrate, polydimethylsiloxane (PDMS), under standard soft lithography. The performance of the microdevice is high-throughput, sensitive, and user-friendly. A total of 53.1% of the triangular microwells were able to trap single canine cutaneous mast cell tumor (MCT) cells. Of these, 38.82% were single cell entrapments, while 14.34% were multiple cell entrapments. The ratio of single-to-multiple cell trapping was high, at 2.7:1. In addition, 80.5% of the trapped cells were viable, indicating that the system was non-lethal. OCT4A-immunofluorescence combined with the proposed system can assess OCT4A expression in trapped single cells more precisely than OCT4A-immunohistochemistry. Therefore, the results suggest that microfluidic single cell analysis could potentially reduce the impact of cellular heterogeneity.

11.
Micromachines (Basel) ; 10(11)2019 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-31726665

RESUMO

Inertial separation techniques in a microfluidic system have been widely employed in the field of medical diagnosis for a long time. Despite no requirement of external forces, it requires strong hydrodynamic forces that could potentially cause cell damage or loss during the separation process. This might lead to the wrong interpretation of laboratory results since the change of structures and functional characteristics of cells due to the hydrodynamic forces that occur are not taken into account. Therefore, it is important to investigate the cell viability and damage along with the separation efficacy of the device in the design process. In this study, two inertial separation techniques-spiral microchannel and contraction-expansion array (CEA)-were examined to evaluate cell viability, morphology and intracellular structures using a trypan blue assay (TB), Scanning Electron Microscopy (SEM) and Wright-Giemsa stain. We discovered that cell loss was not significantly found in a feeding system, i.e., syringe, needle and tube, but mostly occurred in the inertial separation devices while the change of cell morphology and intracellular structures were found in the feeding system and inertial separation devices. Furthermore, percentage of cell loss was not significant in both devices (7-10%). However, the change of cell morphology was considerably increased (30%) in spiral microchannel (shear stress dominated) rather than in CEA (12%). In contrast, the disruption of intracellular structures was increased (14%) in CEA (extensional and shear stress dominated equally) rather than spiral microchannel (2%). In these experiments, leukocytes of canine were used as samples because their sizes are varied in a range between 7-12 µm, and they are commonly used as a biomarker in many clinical and medical applications.

12.
Parasit Vectors ; 12(1): 159, 2019 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-30961652

RESUMO

BACKGROUND: The diagnosis of filariasis traditionally relies on the detection of circulating microfilariae (mf) using Giemsa-stained thick blood smears. This approach has several limitations. We developed a semi-automated microfluidic device to improve and simplify the detection of filarial nematodes. METHODS: The efficiency and repeatability of the microfluidic device was evaluated. Human EDTA blood samples were 'spiked' with B. malayi mf at high, moderate, and low levels, and subsequently tested 10 times. The device was also used for a field survey of feline filariasis in 383 domesticated cats in an area of Narathiwat Province, Thailand, the endemic area of Brugia malayi infection. RESULTS: In the control blood arbitrarily spiked with mf, the high level, moderate level and low level mf-positive controls yielded coefficient variation (CV) values of 4.44, 4.16 and 4.66%, respectively, at the optimized flow rate of 6 µl/min. During the field survey of feline filariasis in Narathiwat Province, the device detected mf in the blood of 34 of 383 cats (8.9%) whereas mf were detected in 28 (7.3%) cats using the blood smear test. Genomic DNA was extracted from mf trapped in the device after which high-resolution melting (HRM) real-time PCR assay was carried out, which enabled the simultaneous diagnosis of filarial species. Among the 34 mf-positive samples, 12 were identified as B. malayi, 15 as Dirofilaria immitis and 7 as| D. repens. CONCLUSIONS: We developed a semi-automated microfluidic device to detect mf of filarial parasites that could be used to diagnose lymphatic filariasis in human populations. This novel device facilitates rapid, higher-throughput detection and identification of infection with filariae in blood samples.


Assuntos
Doenças do Gato/diagnóstico , Filariose/veterinária , Técnicas Analíticas Microfluídicas/veterinária , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Animais , Automação Laboratorial , Gatos , Filariose/diagnóstico , Reprodutibilidade dos Testes
13.
Talanta ; 188: 606-613, 2018 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-30029420

RESUMO

This work reports a novel method for in situ measurement of binding of cobalt ions to polyethyleneimine (PEI) and polyethyleneimine-functionalized poly (methyl methacrylate) nanoparticles (PEI-NPs) using simple microfluidics with a chemiluminescence detection system. The catalytic effect of free cobalt ion in solution on the luminol-hydrogen peroxide chemiluminescence was employed for the detection of unbound cobalt in dynamic equilibrium with cobalt bound to PEI or PEI-NPs. Many binding measurements lead to incorrect estimation of free metal ions due to insufficient separation of bound and free ions. The catalytic activity of only unbound cobalt ion on the luminol reaction was demonstrated by observing that PEI and PEI-NPs alone did not give chemiluminescence. Also, both Co-PEI and Co-PEI-NPs complexes gave no chemiluminescence when cobalt ion is fully bound with excess PEI or PEI-NPs. In addition diethylenetriamine (dien) as a model ligand to completely bind the cobalt ions was also employed as further confirmation. The chemiluminescence measurement employing microfluidics was then successfully applied for the measurement of binding cobalt ion to PEI and PEI-NPs. This in situ measurement of binding does not require filtration of the two species. As there is no perturbation of equilibrium, an accurate binding measurement can therefore be successfully performed. Experimental parameters, such as concentrations of polymers and cobalt ions, and equilibration time were investigated. Analysis of the experimental data employed the binding equation derived assuming independent and equivalent binding sites of the polymer for the metal ions. Also the binding constant of cobalt ions with PEI-NPs is first reported employing chemiluminescence detection. This work provides quantitative determination of the binding constant and total binding capacity of PEI and PEI-NPs with cobalt ions using chemiluminescence detection and microfluidics as an innovative in situ measurement of the unbound cobalt ions.

14.
Anal Sci ; 34(2): 161-167, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29434101

RESUMO

A microfluidic method with front-face fluorometric detection was developed for the determination of total inorganic iodine in drinking water. A polydimethylsiloxane (PDMS) microfluidic device was employed in conjunction with the Sandell-Kolthoff reaction, in which iodide catalyzed the redox reaction between Ce(IV) and As(III). Direct alignment of an optical fiber attached to a spectrofluorometer was used as a convenient detector for remote front-face fluorometric detection. Trace inorganic iodine (IO3- and I-) present naturally in drinking water was measured by on-line conversion of iodate to iodide for determination of total inorganic iodine. On-line conversion efficiency of iodate to iodide using the microfluidic device was investigated. Excellent conversion efficiency of 93 - 103% (%RSD = 1.6 - 11%) was obtained. Inorganic iodine concentrations in drinking water samples were measured, and the results obtained were in good agreement with those obtained by an ICP-MS method. Spiked sample recoveries were in the range of 86%(±5) - 128%(±8) (n = 12). Interference of various anions and cations were investigated with tolerance limit concentrations ranging from 10-6 to 2.5 M depending on the type of ions. The developed method is simple and convenient, and it is a green method for iodine analysis, as it greatly reduces the amount of toxic reagent consumed with reagent volumes in the microfluidic scale.


Assuntos
Água Potável/química , Fluorometria/métodos , Iodo/análise , Dispositivos Lab-On-A-Chip , Dimetilpolisiloxanos/química , Iodatos/química , Iodo/química , Limite de Detecção
15.
Micromachines (Basel) ; 9(1)2017 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-30393286

RESUMO

Our laboratory has the fundamental responsibility to study cancer stem cells (CSC) in various models of human and animal neoplasms. However, the major impediments that spike our accomplishment are the lack of universal biomarkers and cellular heterogeneity. To cope with these restrictions, we have tried to apply the concept of single cell analysis, which has hitherto been recommended throughout the world as an imperative solution pack for resolving such dilemmas. Accordingly, our first step was to utilize a predesigned spiral microchannel fabricated by our laboratory to perform size-based single cell separation using mast cell tumor (MCT) cells as a model. However, the impact of hydrodynamic shear stresses (HSS) on mechanical cell injury and viability in a spiral microchannel has not been fully investigated so far. Intuitively, our computational fluid dynamics (CFD) simulation has strongly revealed the formations of fluid shear stress (FSS) and extensional fluid stress (EFS) in the sorting system. The panel of biomedical assays has also disclosed cell degeneration and necrosis in the model. Therefore, we have herein reported the combinatorically detrimental effect of FSS and EFS on the viability of MCT cells after sorting in our spiral microchannel, with discussion on the possibly pathogenic mechanisms of HSS-induced cell injury in the study model.

16.
Heliyon ; 2(10): e00188, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27822563

RESUMO

ImmunoFET (IMFET) biosensor is a simple platform for the detection of biotinylated products of polymerase chain reaction (PCR). Construction of the IMFET biosensor started with adsorption of 1.5 mg/mL of protein A (PA) onto the insulated gate surface of ISFET for 90 min. Next, the immobilized 1/500 dilution of anti-biotin antibody was adsorbed onto the PA layer for 60 min. The IMFET biosensor was subsequently ready for detection of the biotinylated amplicon. The IMFET biosensor showed highly specific binding to the biotinylated PCR product of the phaE gene of Haloquadratum walsbyi DSM 16854. The phaE gene is a biomarker of polyhydroxyalkanoate (PHA) producers that contain PHA synthase class III. The lowest amount of DNA template of H. walsbyi DSM 16854 that the IMFET biosensor could detect was 125 fg. The IMFET biosensor has a lower amount of detection compared with a DNA lateral flow biosensor from our previous study. The degree of linearity of the biosensor signal was influenced by the concentration of the biotinylated amplicon. The IMFET biosensor also has a short response time (approximately 30 times) to detect the phaE amplicon compared to an agarose gel electrophoresis. The IMFET biosensor is a promising tool for the detection of the biotinylated PCR product, and it can be integrated into a micro total analysis system (µTAS).

17.
Analyst ; 141(20): 5767-5775, 2016 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-27486595

RESUMO

A silicon nitride Ion Sensitive Field Effect Transistor (ISFET) based immunosensor was developed as a low-cost and label-free electrical detection for the detection of antigen 85 complex B (Ag85B). The sensing membrane of the ISFET was modified with 3-aminopropyltriethoxysilane (APTES) followed by glutaraldehyde (GA), yielding an aldehyde-terminated surface. This group is available for immobilization of a monoclonal antibody against a recombinant Ag85B protein (anti-Ag85B antibody). The optimal concentration for anti-Ag85B antibody immobilization onto the modified ISFET was 100 µg ml-1. This optimal condition provided the maximal binding capability and minimal non-specific background signal. The binding event between the recombinant Ag85B antigen and anti-Ag85B antibody on the ISFET surface is presented by monitoring the gate potential change at a constant drain current. The dose response for the recombinant Ag85B protein showed a linear response between 0.12 and 1 µg ml-1 without significant interference from other recombinant proteins. The analytical imprecision (CV%) and accuracy of this Ag85B protein biosensor were 9.73-10.99% and 95.29%, respectively. In addition, an irrelevant antibody and other recombinant proteins were employed as a negative control to demonstrate the non-specific interaction of the antigen and antibody. The success of this immunosensor system for Ag85B protein detection facilitates the construction of a promising device which can shorten the turnaround time for the diagnosis of tuberculosis compared to a standard culture method. Furthermore, this device could also be applied for real-time growth monitoring of Mycobacterium tuberculosis in a mycobacterial culture system.


Assuntos
Aciltransferases/análise , Antígenos de Bactérias/análise , Proteínas de Bactérias/análise , Técnicas Biossensoriais , Compostos de Silício , Tuberculose/diagnóstico , Anticorpos Imobilizados , Anticorpos Monoclonais , Glutaral , Íons , Mycobacterium tuberculosis/crescimento & desenvolvimento , Propilaminas , Silanos
18.
Biosens Bioelectron ; 67: 134-8, 2015 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-25108848

RESUMO

Three different types of surface, silicon dioxide (SiO2), silicon nitride (Si3N4), and titanium oxynitride (TiON) were modified for lactate dehydrogenase (LDH) immobilization using (3-aminopropyl)triethoxysilane (APTES) to obtain an amino layer on each surface. The APTES modified surfaces can directly react with LDH via physical attachment. LDH can be chemically immobilized on those surfaces after incorporation with glutaraldehyde (GA) to obtain aldehyde layers of APTES-GA modified surfaces. The wetting properties, chemical bonding composition, and morphology of the modified surface were determined by contact angle (CA) measurement, Fourier transform infrared (FTIR) spectroscopy, and scanning electron microscopy (SEM), respectively. In this experiment, the immobilized protein content and LDH activity on each modified surface was used as an indicator of surface modification achievement. The results revealed that both the APTES and APTES-GA treatments successfully link the LDH molecule to those surfaces while retaining its activity. All types of tested surfaces modified with APTES-GA gave better LDH immobilizing efficiency than APTES, especially the SiO2 surface. In addition, the SiO2 surface offered the highest LDH immobilization among tested surfaces, with both APTES and APTES-GA modification. However, TiON and Si3N4 surfaces could be used as alternative candidate materials in the preparation of ion-sensitive field-effect transistor (ISFET) based biosensors, including lactate sensors using immobilized LDH on the ISFET surface.


Assuntos
Técnicas Biossensoriais/métodos , Glutaral/química , L-Lactato Desidrogenase/química , Ácido Láctico/química , Compostos de Silício/química , Dióxido de Silício/química , Adsorção , Ativação Enzimática , Estabilidade Enzimática , Enzimas Imobilizadas/química , Ácido Láctico/análise , Teste de Materiais , Ligação Proteica , Propriedades de Superfície
19.
J Nanosci Nanotechnol ; 12(6): 4919-27, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22905552

RESUMO

Needle-shaped pillars so-called "Black silicon" (B-Si) were fabricated by etching cleaned silicon wafer with fluorine-based deep reactive ion etching plasma. The B-Si pillar with the pillar size (a) and spacing (b) of 250 nm, and height (h) of 6.47 microm, coated with SiOxFy film had water contact angle (WCA) and ethylene glycol contact angle (ECA) of 159.8 degrees and 135.5 degrees, respectively. After coating the pillar with trichloro(1H,1H, 2H,2H-perfluorooctyl)silane (TPFS), the WCA and ECA increased to 166.2 degrees and 161.8 degrees, respectively. At the optimum etching condition, the B-Si pillar with the size a = 376 nm, b = 576 nm, h = 6.47microm, and the aspect ratio of 14.80 showed the WCA and ECA of 4.25 degrees and 14.77 degrees, respectively. After coating with the TPFS, liquid droplets ran across the sample's surface rapidly and the WCA and ECA could not be measured. Moreover, when the pillar height was increased twice, the WCA and ECA of the B-Si with and without the TPFS coating were greater than 170 degrees, indicating excellent water-and-oil repellency and can be applied for Micro-Electro-Mechanical Systems (MEMS).


Assuntos
Cristalização/métodos , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Gases em Plasma/química , Silício/química , Interações Hidrofóbicas e Hidrofílicas , Substâncias Macromoleculares/química , Teste de Materiais , Conformação Molecular , Tamanho da Partícula , Propriedades de Superfície
20.
J Nanosci Nanotechnol ; 11(10): 8967-73, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22400288

RESUMO

Superhydrophobic surface can be fabricated by creating a rough surface at very fine scale and modify it with low-surface energy material. To obtain the optimum superhydrophobicity, the surface roughness must be maximized. To avoid the limitation of scaling down the pattern size by using an expensive lithography tools, the surface roughness factor (r) was increased by means of changing an asperity shape so as to increase its overall surface area. In this paper, the patterns of the asperities under studied were wave stripes, line stripes, cylindrical pillars, square pillars, pentagonal pillars, hexagonal pillars, and octagonal pillars. All pillar shapes were arranged in square arrays, hexagonal arrays, and continuous stripes. The asperities sizes and the pitches were varied from 1 to 5 microm with 10 microm of asperity height. Then the patterned surfaces were coated with polydimethylsiloxane mixed with 10 wt% dicumylperoxide. It was found that the stripe asperities can generate only hydrophobic surface with water contact angle (WCA) of 135 degrees to 145 degrees. The pillars with square and hexagonal arrays had the WCA of 149 degrees to 158 degrees. The pentagonal pillars with square and hexagonal arrays achieved the highest WCA with an average WCA of 156 degrees. It was evident that the pillar shape had significant effect on the superhydrophobicity.

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