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10 pages, 1382 KiB  
Article
Variable Angle Spectroscopic Ellipsometry Characterization of DMOAP-Functionalized Graphene Oxide Films
by Grazia Giuseppina Politano and Carlo Versace
Coatings 2023, 13(12), 2066; https://doi.org/10.3390/coatings13122066 - 11 Dec 2023
Cited by 2 | Viewed by 1348
Abstract
In our research, we investigated the optical properties of Graphene Oxide (GO) films functionalized with N,N-Dimethyl-N-octadecyl(3-aminopropyl)trimethoxysilyl chloride (DMOAP) using Variable Angle Spectroscopic Ellipsometry (VASE). We found that after the functionalization process, there was a notable increase in the thickness of the GO films. [...] Read more.
In our research, we investigated the optical properties of Graphene Oxide (GO) films functionalized with N,N-Dimethyl-N-octadecyl(3-aminopropyl)trimethoxysilyl chloride (DMOAP) using Variable Angle Spectroscopic Ellipsometry (VASE). We found that after the functionalization process, there was a notable increase in the thickness of the GO films. Additionally, there were observable changes in the Lorentz oscillator parameters, signaling modifications in the electronic transitions and structural properties of the films. Our results indicate that the functionalization with DMOAP not only impacts the electronic conjugation within the GO structures but also enhances the optical conductivity of the films, which is a finding of significant importance for potential applications in electronics, photonics, and materials science. Full article
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Figure 1

Figure 1
<p>Variable angle spectroscopic ellipsometry of DMOAP-functionalized graphene oxide films on SiO<sub>2</sub>/Si substrates. Comparison of experimental and simulated (<b>a</b>) ψ and (<b>b</b>) Δ at 60°, 65° and 70° incident angles.</p>
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<p>Estimated dispersion laws of the DMOAP-functionalized graphene oxide films. The black curve illustrates the index of refraction (n), while the blue curve represents the extinction coefficient (k).</p>
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<p>Optical conductivity variation with frequency for both (<b>a</b>) graphene oxide films and (<b>b</b>) graphene oxide films treated with DMOAP on SiO<sub>2</sub>/Si substrates.</p>
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11 pages, 1975 KiB  
Article
Aptamer-Based Gold Nanoparticles–PDMS Composite Stamps as a Platform for Micro-Contact Printing
by Amna Didar Abbasi, Zakir Hussain and Kun-Lin Yang
Biosensors 2022, 12(12), 1067; https://doi.org/10.3390/bios12121067 - 23 Nov 2022
Cited by 2 | Viewed by 2711
Abstract
In the present study, a functional template made up of in situ synthesised gold nanoparticles (AuNPs) is prepared on polydimethylsiloxane (PDMS) for patterning of target protein onto the desired solid substrates. Unlike previous studies in which bioreceptor probes are randomly attached to the [...] Read more.
In the present study, a functional template made up of in situ synthesised gold nanoparticles (AuNPs) is prepared on polydimethylsiloxane (PDMS) for patterning of target protein onto the desired solid substrates. Unlike previous studies in which bioreceptor probes are randomly attached to the PDMS stamp through electrostatic interactions, herein, we propose an AuNPs–PDMS stamp, which provides a surface for the attachment of thiol-modified biorecognition probes to link to the stamp surface through a dative bond with a single anchoring point based on thiol chemistry. By using this platform, we have developed the ability for microcontact printing (µCP) to selectively capture and transfer target protein onto solid surfaces for detection purposes. After µCP, we also investigated whether liquid crystals (LCs) could be used as a label-free approach for identifying transfer protein. Our reported approach provides promise for biosensing of various analytes. Full article
(This article belongs to the Special Issue Biosensing and Diagnosis)
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Figure 1

Figure 1
<p>Schematic illustration of the fabrication of AuNPs–PDMS stamp and biorecognition process, including inking with probe aptamer, stamping and patterning target protein gp-120 on DMOAP-coated glass slide through µCP.</p>
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<p>UV-vis absorption spectra. (<b>a</b>) UV-Vis spectra of bare PDMS and AuNPs–PDMS, (<b>b</b>) XRD of AuNPs–PDMS, (<b>c</b>) FeSEM image of bare PDMS and, (<b>d</b>) AuNPs–PDMS composite.</p>
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<p>UV-Vis absorption spectra of AuNPs–PDMS with immobilized biorecognition aptamer probe.</p>
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<p>Fluorescence images showing (<b>a</b>) AuNPs–PDMS stamp surface with a circular region of immobilised RNA probe (3’ 6-FAM-conjugated Apt 8), and (<b>b</b>) DMOAP-coated glass slide surface after µCP. The RNA probe is stable and not transferred from the stamp to the DMOAP-coated glass slide during µCP.</p>
Full article ">Figure 5
<p>Polarized optical images of LCs supported on DMOAP-coated glass slide printed by AuNPs–PDMS stamp with immobilized probe (B40t77 aptamer) incubated with (<b>a</b>) 8 (µg/mL), (<b>b</b>) 6 (µg/mL), (<b>c</b>) 4 (µg/mL), (<b>d</b>) (2 µg/mL), (<b>e</b>) 1 (µg/mL), (<b>f</b>) 0.5 (µg/mL) and (<b>g</b>) (0 µg/mL) of target protein gp-120.</p>
Full article ">Figure 6
<p>Specificity of the AuNPs–PDMS stamp with immobilized B40t77 aptamer probe after incubation with a solution of the mixture of two proteins (FITC-IgG + gp-120) is demonstrated by fluorescence images with no signals show no binding and transfer of FITC-IgG, (<b>a</b>) when 8 µg/mL quantity of gp-120 and FITC-IgG were mixed, and (<b>b</b>) when 8 µg/mL of gp-120 and 16 µg/mL of FITC-IgG were mixed. Bright optical images of LCs showing selective binding and transfer of gp-120 onto DMOAP-coated glass slide, (<b>c</b>) when 8 µg/mL quantity of gp-120 and FITC-IgG were mixed, and (<b>d</b>) when 8 µg/mL of gp-120 and 16 µg/mL of FITC-IgG were mixed.</p>
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13 pages, 3823 KiB  
Article
Quantitative Biosensing Based on a Liquid Crystal Marginally Aligned by the PVA/DMOAP Composite for Optical Signal Amplification
by Tsung-Keng Chang, Mon-Juan Lee and Wei Lee
Biosensors 2022, 12(4), 218; https://doi.org/10.3390/bios12040218 - 7 Apr 2022
Cited by 9 | Viewed by 2601
Abstract
The working principle for a liquid crystal (LC)-based biosensor relies on the disturbance in the orderly aligned LC molecules induced by analytes at the LC-aqueous or LC-solid interface to produce optical signals that can be typically observed under a polarizing optical microscope (POM). [...] Read more.
The working principle for a liquid crystal (LC)-based biosensor relies on the disturbance in the orderly aligned LC molecules induced by analytes at the LC-aqueous or LC-solid interface to produce optical signals that can be typically observed under a polarizing optical microscope (POM). Our previous studies demonstrate that such optical response can be enhanced by imposing a weak electric field on LCs so that they are readily tilted from the homeotropic alignment in response to lower concentrations of analytes at the LC-glass interface. In this study, an alternative approach toward signal amplification is proposed by taking advantage of the marginally tilted alignment configuration without applying an electric field. The surface of glass substrates was modified with a binary aligning agent of poly(vinyl alcohol) (PVA) and dimethyloctadecyl[3-(trimethoxysilyl)propyl] ammonium chloride (DMOAP), in which the amount of PVA was fine-tuned so that the interfacing LC molecules were slightly tilted but remained virtually homeotropically aligned to yield no light leakage under the POM in the absence of an analyte. Two nematic LCs, E7 and 5CB, were each sandwiched between two parallel glass substrates coated with the PVA/DMOAP composite for the detection of bovine serum albumin (BSA), a model protein, and cortisol, a small-molecule steroid hormone. Through image analysis of the optical appearance of E7 observed under the POM, a limit of detection (LOD) of 2.5 × 10−8 μg/mL for BSA and that of 3 × 10−6 μg/mL for cortisol were deduced. Both values are significantly lower than that obtained with only DMOAP as the alignment layers, which correspond to signal amplification of more than six orders of magnitude. The new approach for signal amplification reported in this work enables analytes of a wide range of molecular weights to be detected with high sensitivity. Full article
(This article belongs to the Special Issue Frontiers in Liquid Crystal-Based Biosensors)
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Figure 1

Figure 1
<p>Signal amplification in LC-based biosensing through marginally tilted alignment of LCs achieved by the aligning layers of the PVA/DMOAP composite. Compared with the DMOAP-coated surface, LC molecules are in a tilted state on the PVA/DMOAP composite, which enables a greater extent of disturbance to occur in the presence of macromolecules such as BSA or small molecules such as cortisol, leading to enhanced optical signal. Blue arrows stand for the direction of light transmission.</p>
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<p>Optical texture of E7 on glass substrates coated with the PVA/DMOAP composite. The nematic LC E7 was sandwiched between two glass substrates coated with PVA/DMOAP mixtures of various mass ratios (2.4:1, 2.6:1, 2.8:1, 3:1, 3.2:1, and 4:1). Scale bar: 500 μm.</p>
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<p>Water contact angle measured on glass substrates coated with the PVA/DMOAP composite. Glass substrates were coated with PVA/DMOAP mixtures of various mass ratios (0:1, 2.4:1, 2.6:1, 2.8:1, 16:1 and 1:0) and subjected to water contact angle measurements. Representative images of the water droplet at each PVA/DMOAP mass ratio are shown at the top of the chart. Error bars represent standard deviations calculated from at least three independent measurements.</p>
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<p>Average tilt angle of E7 as determined by capacitance measurements. The nematic LC E7 was sandwiched between two ITO-coated glass substrates covered with the PVA/DMOAP aligning agent. The average tilt angle was calculated by Equation (1) and plotted against the PVA/DMOAP mass ratio. Error bars represent standard deviations calculated from at least three independent measurements. Corresponding conoscopic micrographs at distinct PVA/DMOAP mass ratios are displayed for comparison.</p>
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<p>Optical texture of E7 and 5CB at various concentrations of BSA on glass substrates coated with the PVA/DMOAP composite. Aqueous solutions of BSA at various concentrations (10<sup>−8</sup> to 10<sup>2</sup> μg/mL) were immobilized on PVA/DMOAP-coated glass substrates for LC-based detection with (<b>a</b>) E7 and (<b>b</b>) 5CB as the sensing media. (<b>c</b>) A parallel study on DMOAP-coated glass substrates was undertaken with a BSA concentration range of 10<sup>0</sup>–10<sup>2</sup> μg/mL. Scale bar: 500 μm.</p>
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<p>Quantitative analysis of the optical responses of E7 (open squares) and 5CB (open triangles) at various concentrations of BSA on glass substrates coated with the PVA/DMOAP composite. The relative intensity of the brightness of the optical texture in <a href="#biosensors-12-00218-f005" class="html-fig">Figure 5</a>a,b were determined by the image processing program ImageJ and plotted against the logarithmic BSA concentration. The linear curve was illustrated in the inset to calculate the LOD. Error bars represent standard deviations calculated from at least three independent experiments. The dotted line represents the fitted regression line, with the determination coefficient R<sup>2</sup> given in the legend.</p>
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<p>Optical texture of (<b>a</b>) E7 and (<b>b</b>) 5CB at various concentrations of cortisol on glass substrates coated with the PVA/DMOAP composite. Aqueous solutions of cortisol at various concentrations (10<sup>−6</sup>–10<sup>−1</sup> μg/mL) were immobilized on PVA/DMOAP-coated glass substrates for LC-based detection with E7 or 5CB as the sensing medium. Scale bar: 500 μm.</p>
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<p>Quantitative analysis of the optical responses of E7 (open squares) and 5CB (open triangles) at various concentrations of cortisol on glass substrates coated with the PVA/DMOAP composite. The relative intensity of the brightness of the optical texture in <a href="#biosensors-12-00218-f007" class="html-fig">Figure 7</a>a,b were determined by the ImageJ software and plotted against the logarithm of cortisol concentration. The linear curve was illustrated in the inset to calculate the LOD. Error bars represent standard deviations calculated from at least three independent experiments. The dotted curve denotes the fitted regression line, with the coefficient of determination, R<sup>2</sup>, shown in the legend.</p>
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14 pages, 3441 KiB  
Article
A Single-Substrate Biosensor with Spin-Coated Liquid Crystal Film for Simple, Sensitive and Label-Free Protein Detection
by Po-Chang Wu, Chao-Ping Pai, Mon-Juan Lee and Wei Lee
Biosensors 2021, 11(10), 374; https://doi.org/10.3390/bios11100374 - 6 Oct 2021
Cited by 8 | Viewed by 2658
Abstract
A liquid crystal (LC)-based single-substrate biosensor was developed by spin-coating an LC thin film on a dimethyloctadecyl[3-(trimethoxysilyl)propyl]ammonium chloride (DMOAP)-decorated glass slide. Compared with the conventional sandwiched cell configuration, the simplified procedure for the preparation of an LC film allows the film thickness to [...] Read more.
A liquid crystal (LC)-based single-substrate biosensor was developed by spin-coating an LC thin film on a dimethyloctadecyl[3-(trimethoxysilyl)propyl]ammonium chloride (DMOAP)-decorated glass slide. Compared with the conventional sandwiched cell configuration, the simplified procedure for the preparation of an LC film allows the film thickness to be precisely controlled by adjusting the spin rate, thus eliminating personal errors involved in LC cell assembly. The limit of detection (LOD) for bovine serum albumin (BSA) was lowered from 10−5 g/mL with a 4.2-μm-thick sandwiched cell of the commercial LC E7 to 107 g/mL with a 4.2-μm-thick spin-coated E7 film and further to 108 g/mL by reducing the E7 film thickness to 3.4 μm. Moreover, by exploiting the LC film of the highly birefringent nematic LC HDN in the immunodetection of the cancer biomarker CA125, an LOD comparable to that determined with a sandwiched HDN cell was achieved at 10−8 g/mL CA125 using a capture antibody concentration an order of magnitude lower than that in the LC cell. Our results suggest that employing spin-coated LC film instead of conventional sandwiched LC cell provides a more reliable, reproducible, and cost-effective single-substrate platform, allowing simple fabrication of an LC-based biosensor for sensitive and label-free protein detection and immunoassay. Full article
(This article belongs to the Special Issue Electrical and Electro-Optical Biosensors)
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Figure 1
<p>Schematic of the optical setup with crossed polarizers for the measurement of the thickness of LC films spin-coated on a DMOAP-coated glass substrate.</p>
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<p>Schematic of the working principle of the LC-based single-substrate protein assay for the detection of BSA with spin-coated LC film on a DMOAP-coated substrate. (<b>a</b>) In the absence of BSA, LC molecules are aligned homeotropically, resulting in uniformly dark optical texture. (<b>b</b>) In the presence of BSA, LC alignment is disrupted at the LC‒glass interface, giving rise to non-uniform and bright optical texture.</p>
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<p>Polarized optical textures of E7 in spin-coated films and LC cells in the presence of BSA. The nematic LC E7 was (<b>a</b>) spin-coated as a thin film on a DMOAP-coated glass substrate or (<b>b</b>) sandwiched between two glass substrates in an LC cell at various BSA concentrations ranging from 10<sup>−7</sup> to 10<sup>−4</sup> g/mL. Scale bar, 500 μm.</p>
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<p>Correlation between spin rate and LC film thickness. The E7 film was spin-coated on DMOAP substrates with a two-step procedure in which the spin rate of the first step <span class="html-italic">ω</span><sub>1</sub> = 1000, 3000 or 5000 rpm and that of the second step <span class="html-italic">ω</span><sub>2</sub> = 5000 rpm, with each step lasting for 10 s. (<b>a</b>) Conoscopic images of LC films spread at various <span class="html-italic">ω</span><sub>1</sub> observed under a POM. Each error bar denotes the standard deviation calculated from the transmission values of five independent measurements. (<b>b</b>) Dependence of transmittance on incident angle of light <span class="html-italic">θ</span> measured with the optical setup in <a href="#biosensors-11-00374-f001" class="html-fig">Figure 1</a>. The values of E7 film thickness displayed in the inset of (<b>b</b>) were deduced by fitting the experimental results with Equation (1).</p>
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<p>Polarized optical textures of spin-coated E7 films of various thicknesses in the presence of BSA. DMOAP-coated glass substrates with immobilized BSA at concentrations of (<b>a</b>) 10<sup>−8</sup>, (<b>b</b>) 10<sup>−7</sup>, and (<b>c</b>) 10<sup>−6</sup> g/mL were spin-coated with the nematic LC E7 by varying the spin rate of the first step (<span class="html-italic">ω</span><sub>1</sub>) of a two-step spin-coating procedure to form LC films with thicknesses <span class="html-italic">d</span><sub>LC</sub> of 4.8, 4.2, and 3.4 μm at <span class="html-italic">ω</span><sub>1</sub> = 1000, 3000, and 5000 rpm, respectively. (<b>d</b>) Optical textures of three independent sets of experiments for 3.4 μm-thick E7 films in the presence of BSA. Note that the brightness of optical textures in <a href="#biosensors-11-00374-f005" class="html-fig">Figure 5</a>a,b,d has been artificially increased by 40% to enhance the visibility. Scale bar, 500 μm.</p>
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<p>Schematics of the working principle of the LC-based single-substrate immunoassay for the detection of CA125 with spin-coated LC film on a DMOAP-coated substrate. (<b>a</b>) In the absence of CA125, LC molecules are aligned homeotropically on immobilized anti-CA125 antibody, whose amount is adjusted to a maximum without disturbing the orientation of LCs. (<b>b</b>) In the presence of CA125, alignment of LC molecules is disrupted due to the formation of the CA125 immunocomplex atop the DMOAP aligning layer.</p>
Full article ">Figure 7
<p>Polarized optical textures of spin-coated E7 and HDN films in the presence of the anti-CA125 antibody. The nematic LCs (<b>a</b>) E7 and (<b>b</b>) HDN were spin-coated on DMOAP-coated substrates immobilized with the anti-CA125 antibody at concentrations ranging from 10<sup>−8</sup> to 10<sup>−5</sup> g/mL. Scale bar, 500 μm.</p>
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<p>Polarized optical textures of spin-coated E7 films in the presence of the CA125 protein. The nematic LC E7 was spin-coated on DMOAP-coated substrates immobilized with 10<sup>−7</sup> g/mL anti-CA125 antibody and reacted with CA125 at concentrations of (<b>a</b>) 10<sup>−6</sup>, (<b>b</b>) 10<sup>−5</sup>, and (<b>c</b>) 10<sup>−4</sup> g/mL. Each red dashed circle represents the area within which the anti-CA125 antibody was immobilized. Scale bar, 500 μm.</p>
Full article ">Figure 9
<p>Polarized optical textures of spin-coated HDN films in the presence of the CA125 protein. The nematic LC HDN was spin-coated on DMOAP-coated substrates immobilized with 10<sup>−8</sup> g/mL anti-CA125 antibody and reacted with CA125 at concentrations of (<b>a</b>) 10<sup>−9</sup>, (<b>b</b>) 10<sup>−8</sup>, (<b>c</b>) 10<sup>−7</sup>, (<b>d</b>) 10<sup>−6</sup>, and (<b>e</b>) 10<sup>−5</sup> g/mL. Each red dashed circle represents the area within which the anti-CA125 antibody was immobilized. Sets 1–3 represent results of three independent experiments. Scale bar, 500 μm.</p>
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7 pages, 1661 KiB  
Communication
Label-Free, Color-Indicating, Polarizer-Free Dye-Doped Liquid Crystal Microfluidic Polydimethylsiloxane Biosensing Chips for Detecting Albumin
by Fu-Lun Chen, Hui-Tzung Luh and Yu-Cheng Hsiao
Polymers 2021, 13(16), 2587; https://doi.org/10.3390/polym13162587 - 4 Aug 2021
Cited by 9 | Viewed by 2037
Abstract
We reveal a novel design for dye-doped liquid crystal (DDLC) microfluidic biosensing chips in the polydimethylsiloxane material. With this design chip, the orientation of DDLCs was affected by the interface between the walls of the channels and DDLCs. When the inside of a [...] Read more.
We reveal a novel design for dye-doped liquid crystal (DDLC) microfluidic biosensing chips in the polydimethylsiloxane material. With this design chip, the orientation of DDLCs was affected by the interface between the walls of the channels and DDLCs. When the inside of a channel was coated with an N,N-dimethyl-n-octadecyl-3-aminopropyltrimethoxysilyl chloride (DMOAP) alignment layer, the DDLCs oriented homeotropically in a homeotropic (H) state under cross-polarized microscopy. After immobilization of antigens with antibodies on the alignment layer-coated microchannel walls, the optical intensity of the DDLC change from the dark H state to the bright planar (P) state. Using pressure-driven flow, the binding of antigens/antibodies to the DDLCs could be detected in an experimental sequential order. The microfluidic DDLCs were tested by detecting bovine serum albumin (BSA) and its immune-responses of antigens/antibodies. We proved that this immunoassay chip was able to detect BSA antigens/antibodies pairs with the detection limit about 0.5 µg/mL. The novel DDLC chip was shown to be a simple, multi-detection device, and label-free microfluidic chips are presented. Full article
(This article belongs to the Special Issue High-Performance Polymeric Sensors II)
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Figure 1
<p>Schematic of microfluidic dye-doped liquid crystal (DDLC) biosensor chips in the presence of bovine serum albumin (BSA) biomolecules.</p>
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<p>Optical mechanism and images from a non-polarized optical microscope of dye-doped liquid crystal (DDLC) microfluidic chips of a biosensor in both the presence and absence of biomolecules.</p>
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<p>Non-polarized optical images of a dye-doped liquid crystal (DDLC) microfluidic biosensor. The bovine serum albumin (BSA) immobilized at concentrations 0~1 mg/mL.</p>
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<p>The linear correlations of the intensity of dye-doped liquid crystal (DDLC) multi-microfluidic chips at various concentrations of bovine serum albumin (BSA).</p>
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<p>The intensities of immunodetection dye-doped liquid crystal (DDLC) multi-microfluidic chips immobilized with concentrations of 0–10 µg/mL bovine serum albumin (BSA) and concentrations of 0–1000 µg/mL of the BSA antibody.</p>
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<p>Intensities of dye-doped liquid crystal (DDLC) biosensor chips at various bovine serum albumin (BSA) with a 1 µg/mL of BSA antibody.</p>
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12 pages, 3729 KiB  
Article
Aptamer Laden Liquid Crystals Biosensing Platform for the Detection of HIV-1 Glycoprotein-120
by Amna Didar Abbasi, Zakir Hussain and Kun-Lin Yang
Molecules 2021, 26(10), 2893; https://doi.org/10.3390/molecules26102893 - 13 May 2021
Cited by 13 | Viewed by 2683
Abstract
We report a label-free and simple approach for the detection of glycoprotein-120 (gp-120) using an aptamer-based liquid crystals (LCs) biosensing platform. The LCs are supported on the surface of a modified glass slide with a suitable amount of B40t77 aptamer, allowing the LCs [...] Read more.
We report a label-free and simple approach for the detection of glycoprotein-120 (gp-120) using an aptamer-based liquid crystals (LCs) biosensing platform. The LCs are supported on the surface of a modified glass slide with a suitable amount of B40t77 aptamer, allowing the LCs to be homeotropically aligned. A pronounced topological change was observed on the surface due to a specific interaction between B40t77 and gp-120, which led to the disruption of the homeotropic alignment of LCs. This results in a dark-to-bright transition observed under a polarized optical microscope. With the developed biosensing platform, it was possible to not only identify gp-120, but obtained results were analyzed quantitatively through image analysis. The detection limit of the proposed biosensing platform was investigated to be 0.2 µg/mL of gp-120. Regarding selectivity of the developed platform, no response could be detected when gp-120 was replaced by other proteins, such as bovine serum albumin (BSA), hepatitis A virus capsid protein 1 (Hep A VP1) and immunoglobulin G protein (IgG). Due to attributes such as label-free, high specificity and no need for instrumental read-out, the presented biosensing platform provides the potential to develop a working device for the quick detection of HIV-1 gp-120. Full article
(This article belongs to the Special Issue Liquid Crystals 2020)
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>Schematic illustration of the preparation and sensing strategy of proposed bio-sensing approach: (<b>A</b>) cleaning and preparation of <span class="html-italic">N,N</span>-dimethyl-n-octadecyl-3aminopropyltrimethoxy-silyl chloride (DMOAP)-coated glass slides; (<b>B</b>) dispensing droplets of B40t77 aptamer solution in an array form; (<b>C</b>) after immobilization of B40t77 aptamer, gp-120 protein droplets were dispensed on immobilized B40t77 aptamer; (<b>D</b>) after washing and drying, place spacer (6µm) on left and right edges of slides, followed by covering the slide with another DMOAP–coated glass slide; (<b>E</b>) clip the left and right edges of both slides with clippers to make an optical cell; (<b>F</b>) introduce liquid crystal (LC) 4-cyano-4-pentylbiphenyl (5CB) into the prepared optical cell; (<b>G</b>) visualization under polarized optical microscope with the naked eye, analysis of final results (dark to bright transition in optical appearances).</p>
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<p>Optical images of LCs supported on DMOAP-coated surface immobilized with B40t77 aptamer solutions made in different MgCl<sub>2</sub> concentrations; MgCl<sub>2</sub> concentrations are 1 M, 100 mM, 10 mM and 0 mM; control is just TE buffer with different concentrations of MgCl<sub>2</sub>.</p>
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<p>Optical images of LCs supported on a DMOAP-coated slide immobilized with B40t77 aptamer solution with different concentrations; number above each spot is B40t77 aptamer concentration (µg/mL), whereas control is just TE buffer without B40t77 aptamer.</p>
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<p>Optical images of LCs supported on a DMOAP-coated slide immobilized with B40t77 aptamer solution of different concentrations showing effects of different immobilization times; the B40t77 aptamer concentrations used are 16, 14, 12, 10 and 8 µg/mL.</p>
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<p>AFM images of different slide surfaces: (<b>A</b>) with DMOAP coating; (<b>B</b>) after B40t77 aptamer (8 µg/mL) immobilization; (<b>C</b>) after addition of gp-120 (8 µg/mL), and the corresponding polarized optical images of LCs supported on different slide surfaces; (<b>D</b>) coated with DMOAP; (<b>E</b>) with B40t77 aptamer (8 µg/mL) modification; (<b>F</b>) after addition of gp-120 (8 µg/mL).</p>
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<p>Optical images of LCs supported on B40t77 aptamer (8 µg/mL) immobilized DMOAP-coated glass surfaces after addition of different concentrations of gp-120: (<b>A</b>) 8, (<b>B</b>) 6, (<b>C</b>) 4, (<b>D</b>) 2, (<b>E</b>) 1, (<b>F</b>) 0.5, (<b>G</b>) 0.2, (<b>H</b>) 0.1 and (<b>I</b>) 0 µg/mL, respectively.</p>
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<p>(<b>A</b>–<b>I</b>) are showing mean gray values corresponding to different optical images of LCs given in <a href="#molecules-26-02893-f006" class="html-fig">Figure 6</a>.</p>
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<p>Calibration curve of the mean gray values of LC optical images and the logarithm of the gp-120 concentration (Y = mean gray value of the optical images and C<sub>gp-120</sub> = concentration of gp-120 (ng/mL)).</p>
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<p>Optical images of LCs supported on B40t77 immobilized DMOAP decorated glass surfaces after treating with different proteins at concentration (8 µg/mL): (<b>A</b>) Gp-120, (<b>B</b>) BSA, (<b>C</b>) Hep A VP1, (<b>D</b>) IgG and (<b>E</b>–<b>G</b>) optical images with double concentration (16 µg/mL) of IgG, Hep A VP1 and BSA, respectively.</p>
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12 pages, 3828 KiB  
Article
A Label-Free Liquid Crystal Biosensor Based on Specific DNA Aptamer Probes for Sensitive Detection of Amoxicillin Antibiotic
by Duy Khiem Nguyen and Chang-Hyun Jang
Micromachines 2021, 12(4), 370; https://doi.org/10.3390/mi12040370 - 30 Mar 2021
Cited by 26 | Viewed by 4296
Abstract
We developed a liquid crystal (LC) aptamer biosensor for the sensitive detection of amoxicillin (AMX). The AMX aptamer was immobilized onto the surface of a glass slide modified with a mixed self-assembled layer of dimethyloctadecyl [3-(trimethoxysilyl) propyl] ammonium chloride (DMOAP) and (3-aminopropyl) triethoxysilane [...] Read more.
We developed a liquid crystal (LC) aptamer biosensor for the sensitive detection of amoxicillin (AMX). The AMX aptamer was immobilized onto the surface of a glass slide modified with a mixed self-assembled layer of dimethyloctadecyl [3-(trimethoxysilyl) propyl] ammonium chloride (DMOAP) and (3-aminopropyl) triethoxysilane (APTES). The long alkyl chains of DMOAP maintained the LC molecules in a homeotropic orientation and induced a dark optical appearance under a polarized light microscope (POM). In the presence of AMX, the specific binding of the aptamer and AMX molecules induced a conformational change in the aptamers, leading to the disruption of the homeotropic orientation of LCs, resulting in a bright optical appearance. The developed aptasensor showed high specificity and a low detection limit of 3.5 nM. Moreover, the potential application of the developed aptasensor for the detection of AMX in environmental samples was also demonstrated. Therefore, the proposed aptasensor is a promising platform for simple, rapid, and label-free monitoring of AMX in an actual water environment with high selectivity and sensitivity. Full article
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Figure 1
<p>Schematic illustration of the sensing strategy for detecting amoxicillin (AMX) (<b>a</b>) Cleaned glass slide; (<b>b</b>) upper glass slide; (<b>c</b>) the self-assembled monolayer of (3-aminopropyl) triethoxysilane (APTES)/dimethyloctadecyl [3-(trimethoxysilyl) propyl] ammonium chloride (DMOAP) on the glass substrate; (<b>d</b>) grafting of glutaraldehyde (GA) onto APTES/DMOAP-modified glass substrate; (<b>e</b>) immobilization of the AMX aptamer onto (APTES + GA)/DMOAP-modified glass substrate; (<b>f</b>) homeotropic orientation of liquid crystals (LCs) in the absence of AMX; (<b>g</b>) polarized optical image of an LC cell in the absence of AMX; (<b>h</b>) the conformational change in AMX aptamer after binding to AMX disrupts the homeotropic orientation of LCs; and (<b>i</b>) polarized optical image of an LC cell in the presence of AMX. The bright circular spot is caused by the specific binding of aptamer and AMX.</p>
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<p>Polarized optical (POM) images of LC cells with 5CB after the modification of the surface of the bottom glass slide with various concentrations of AMX aptamer: (<b>a</b>) 1 µM; (<b>b</b>) 750 nM; (<b>c</b>) 500 nM; (<b>d</b>) 250 nM; (<b>e</b>) 200 nM; and (<b>f</b>) 100 nM. The circular spot shown in the optical images is caused by the presence of AMX aptamers. Scale bar, 250 µm.</p>
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<p>POM images of LC cells with AMX at the concentration of: (<b>a</b>) 1 µM; (<b>b</b>) 800 nM; (<b>c</b>) 600 nM; (<b>d</b>) 400 nM; (<b>e</b>) 200 nM; (<b>f</b>) 100 nM; (<b>g</b>) 50 nM; (<b>h</b>) 10 nM; (<b>i</b>) 5 nM; and (<b>j</b>) 0 nM. Scale bar, 250 µm.</p>
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<p>Three-dimensional AFM images of substrates immobilized with 200 nM AMX aptamer (<b>a</b>), and upon incubation of 800 nM amoxicillin (<b>b</b>).</p>
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<p>Calibration curve of the average gray-scale intensities (GIs) of the POM images vs. the concentration of AMX (y is the average GI of the POM image, x is the concentration of AMX, R<sup>2</sup> is the linear correlation coefficient) (<span class="html-italic">n</span> = 3).</p>
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<p>POM images of LC cells in which the bottom glass substrates were incubated with different antibiotic solutions: (<b>a</b>) 0.8 µM AMX; (<b>b</b>) 1 µM SMXZ; (<b>c</b>) 1 µM TC; (<b>d</b>) 1 µM CL; (<b>e</b>) 1 µM NRFX; (<b>f</b>) 1 µM AMP; (<b>g</b>) 1 µM SMZ; and (<b>h</b>) 1 µM PC. (<b>i</b>) Average GI of optical images caused by amoxicillin (0.8 µM) and other control antibiotics (1 µM). Scale bar, 250 µm.</p>
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12 pages, 7100 KiB  
Article
Bovine Serum Albumin Protein-Based Liquid Crystal Biosensors for Optical Detection of Toxic Heavy Metals in Water
by Noor ul Amin, Humaira Masood Siddiqi, Yang Kun Lin, Zakir Hussain and Nasir Majeed
Sensors 2020, 20(1), 298; https://doi.org/10.3390/s20010298 - 5 Jan 2020
Cited by 34 | Viewed by 6072
Abstract
A new methodology involving the use of Bovine Serum Albumin (BSA) as a probe and liquid crystal (LC) as a signal reporter for the detection of heavy metal ions in water at neutral pH was developed. BSA acted as a multi-dentate ligand for [...] Read more.
A new methodology involving the use of Bovine Serum Albumin (BSA) as a probe and liquid crystal (LC) as a signal reporter for the detection of heavy metal ions in water at neutral pH was developed. BSA acted as a multi-dentate ligand for the detection of multiple metal ions. The LC sensor was fabricated by immobilizing 3 µg mL−1 BSA solution on dimethyloctadecyl-[3-(trimethoxysilyl)propyl]ammonium chloride (DMOAP)-coated glass slides. In the absence of heavy metal ions, a dark optical image was observed, while in their presence, a dark optical image turned to bright. The optical response was characterized by using a polarized optical microscope (POM). The BSA based LC sensor selectively detected toxic metal ions as compared to s block metal ions and ammonium ions in water. Moreover, the limit of detection was found to be very low (i.e., 1 nM) for the developed new biosensor in comparison to reported biosensors. Full article
(This article belongs to the Section Biosensors)
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Graphical abstract

Graphical abstract
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<p>The optical results of an immobilized mixture of 1 mg mL<sup>−1</sup> Bovine Serum Albumin (BSA) and 5 μΜ Cu<sup>2+</sup> on (<b>a</b>) UV-modified N,N-Dimethyl-N-octadecyl-3-Aminopropyltrimethoxyysilychloride (DMOAP) surface (<b>b</b>) unmodified DMOAP surfaces (<b>sd</b>) Schematic diagram of the immobilized mixture.</p>
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<p>Optical images of liquid crystal (LC) of 5 µΜ (<b>a</b>–<b>d</b>) and 10 nM (<b>e</b>) of Cu<sup>2+</sup> mixed with various concentrations of BSA immobilized on modified DMOAP surfaces. Whereas the optical image (<b>f</b>) has been taken without Cu<sup>2+</sup>.</p>
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<p>Schematic representation of detection mechanism of the sensing system designed for the detection of metal ions (<b>a</b>) DMOAP-coated glass slide polarized optical microscope (POM) dark image (<b>b</b>) Immobilized (3 μg mL<sup>−1</sup>) BSA without metal ions on a DMOAP-coated glass slide POM image showing negligible distortion in LC pattern as this amount is soluble in water and is uniformly distributed on DMOAP and (<b>c</b>) BSA with metal ions on DMOAP-coated glass slide showing distorted alignment of LC induced by BSA + metal aggregation showing bright POM image.</p>
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<p>BSA probe shows the selective detection of heavy metals.</p>
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<p>Optical images of LC immobilized mixtures of various Cu<sup>2+</sup> ion concentration with 3 μg mL<sup>−1</sup> BSA.</p>
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<p>Optical images of LC supported on surfaces with immobilized mixtures of different concentrations of Pb<sup>2+</sup> ions with 3 μg mL<sup>−1</sup>of BSA.</p>
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<p>Images show the limit of detection of heavy metals ions with 3 μg mL<sup>−1</sup> of BSA.</p>
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<p>The optical textures of immobilized mixture of 3 μg mL<sup>−1</sup> BSA and Cu(NO<sub>3</sub>)<sub>2</sub>, Cu(ClO<sub>4</sub>)<sub>2</sub> and CuSO<sub>4</sub> at various concentrations.</p>
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<p>Optical images of LC of immobilized mixtures of Pb<sup>2+</sup> (<b>a</b>) 0 µM (<b>b</b>) 0.5 µM and (<b>c</b>) 20 µM with 3 μg mL<sup>−1</sup> BSA.</p>
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