Electrokinetics-Based Microfluidic Technology for the Rapid Separation and Concentration of Bacteria/Cells/Biomolecules

2014 ◽  
Vol 911 ◽  
pp. 347-351 ◽  
Author(s):  
I Fang Cheng ◽  
Tzu Ying Chen ◽  
Hsien Chang Chang
Keyword(s):  
Cancer Cell ◽  
Electrochemical Impedance ◽  
High Sensitivity ◽  
High Specificity ◽  
Cell Detection ◽  
Label Free ◽  
Rapid Separation ◽  
Whole Cells ◽  
Highly Sensitive ◽  
Label Free Detection

Conventional techniques for detection of bacteria/cell and assessment of cancer cell typically use DNA techniques, Western blot and ELISA kits that are high cost, complicated processes and long time consuming. Our researches focus on rapid, portable, simple and highly sensitive separation and detection of cells/bacteria/biomolecules for field-use diagnosis. An ideal portable biosensor (molecular or whole cells detections) unit must have several important features: rapid detection time (<10 minutes), high sensitivity (pM level for molecular detection, 103 cells/ml for whole cell detection), high specificity, small and inexpensive instrumentation configuration. Electrochemical impedance/conductance sensing is preferred over optical detection because of cost and portability concerns. Cancer cell detection using heterogeneous medical samples require continuous isolation, sorting, and trapping of the target bioparticles and immunocolloids within a diagnostic chip. We have developed several electrokinetic strategies to rapid separation, concentration and detection of cells/bacteria/biomolecules in a microfluidic chip using such as dielectrophoresis (DEP), traveling-wave dielectrophoresis (twDEP) and electrohydrodynamics (EHD). Several key techniques we done, which on a rapid/simple/label-free detection platform for the highly sensitive on-chip separation/identification/quantification will be introduced in this paper.

scholarly journals Impedimetric Immunosensor Utilizing Polyaniline/Gold Nanocomposite-Modified Screen-Printed Electrodes for Early Detection of Chronic Kidney Disease

Sensors ◽  
2019 ◽  
Vol 19 (18) ◽  
pp. 3990 ◽  
Author(s):  
Muhammad Omar Shaikh ◽  
Boyanagunta Srikanth ◽  
Pei-Yu Zhu ◽  
Cheng-Hsin Chuang
Keyword(s):  
Chronic Kidney Disease ◽  
Kidney Disease ◽  
Early Detection ◽  
Electrochemical Impedance ◽  
High Specificity ◽  
Homogeneous Distribution ◽  
Label Free ◽  
Gold Nanocrystals ◽  
Working Electrode ◽  
Label Free Detection

The presence of small amounts of human serum albumin (HSA) in urine or microalbuminuria (30–300 µg/mL) is a valuable clinical biomarker for the early detection of chronic kidney disease (CKD). Herein, we report on the development of an inexpensive and disposable immunosensor for the sensitive, specific, and label-free detection of HSA using electrochemical impedance spectroscopy (EIS). We have utilized a simple one-step screen-printing protocol to fabricate the carbon-based three-electrode system on flexible plastic substrates. To enable efficient antibody immobilization and improved sensitivity, the carbon working electrode was sequentially modified with electropolymerized polyaniline (PANI) and electrodeposited gold nanocrystals (AuNCs). The PANI matrix serves as an interconnected nanostructured scaffold for homogeneous distribution of AuNCs and the resulting PANI/AuNCs nanocomposite synergically improved the immunosensor response. The PANI/AuNCs-modified working electrode surface was characterized using scanning electron microscopy (SEM) and the electrochemical response at each step was analyzed using EIS in a ferri/ferrocyanide redox probe solution. The normalized impedance variation during immunosensing increased linearly with HSA concentration in the range of 3–300 µg/mL and a highly repeatable response was observed for each concentration. Furthermore, the immunosensor displayed high specificity when tested using spiked sample solutions containing different concentrations of actin protein and J82 cell lysate (a complex fluid containing a multitude of interfering proteins). Consequently, these experimental results confirm the feasibility of the proposed immunosensor for early diagnosis and prognosis of CKD at the point of care.

scholarly journals Evaluation of Three Peptide Immobilization Techniques on a QCM Surface Related to Acetaldehyde Responses in the Gas Phase

Sensors ◽  
2018 ◽  
Vol 18 (11) ◽  
pp. 3942 ◽  
Author(s):  
Tomasz Wasilewski ◽  
Bartosz Szulczyński ◽  
Wojciech Kamysz ◽  
Jacek Gębicki ◽  
Jacek Namieśnik
Keyword(s):  
Spin Coating ◽  
Olfactory Receptors ◽  
High Sensitivity ◽  
Dip Coating ◽  
Label Free ◽  
Optimal Method ◽  
Spin Coating Method ◽  
Whole Cells ◽  
Label Free Detection ◽  
Coating Method

The quartz-crystal microbalance is a sensitive and universal tool for measuring concentrations of various gases in the air. Biochemical functionalization of the QCM electrode allows a label-free detection of specific molecular interactions with high sensitivity and specificity. In addition, it enables a real-time determination of its kinetic rates and affinity constants. This makes QCM a versatile bioanalytical screening tool for various applications, with surface modifications ranging from the detection of single molecular monolayers to whole cells. Various types of biomaterials, including peptides mapping the binding sites of olfactory receptors, can be deposited as a sensitive element on the surface of the electrodes. One of key ways to ensure the sensitivity and accuracy of the sensor is provided by application of an optimal and repeatable method of immobilization. Therefore, effective sensors operation requires development of an optimal method of deposition. This paper reviews popular techniques (drop-casting, spin-coating, dip-coating) for coating peptides on piezoelectric crystals surface. Peptide (LEKKKKDC-NH2) derived from an aldehyde binding site in the HarmOBP7 protein was synthesized and used as a sensing material for the biosensor. The degree of deposition of the sensitive layer was monitoring by variations in the sensors frequency. The highest mass threshold for QCM measurements for peptides was approximately 16.43 µg·mm−2 for spin coating method. Developed sensor exhibited repeatable response to acetaldehyde. Moreover, responses to toluene was observed to evaluate sensors specificity. Calibration curves of the three sensors showed good determination coefficients (R2 > 0.99) for drop casting and dip coating and 0.97 for the spin-coating method. Sensors sensitivity vs. acetaldehyde were significantly higher for the dip-coating and drop-casting methods and lower for spin-coating one.

Mannosyl electrochemical impedance cytosensor for label-free MDA-MB-231 cancer cell detection

2018 ◽  
Vol 116 ◽  
pp. 100-107 ◽  
Author(s):  
Yi-Hsuan Tang ◽  
Han-Chen Lin ◽  
Chiao-Ling Lai ◽  
Po-Yu Chen ◽  
Chian-Hui Lai
Keyword(s):  
Cancer Cell ◽  
Electrochemical Impedance ◽  
Cell Detection ◽  
Label Free ◽  
Cancer Cell Detection

Design of Photonic Crystals Waveguide Using Microfluidic Infiltration

2014 ◽  
Vol 492 ◽  
pp. 301-305 ◽  
Author(s):  
Faida Bougriou ◽  
Touraya Boumaza ◽  
Mohamed Bouchemat
Keyword(s):  
Photonic Crystals ◽  
Fdtd Method ◽  
High Sensitivity ◽  
Slab Waveguide ◽  
Label Free ◽  
Single Chip ◽  
Photonic Crystal Slab ◽  
Sensing Applications ◽  
Highly Sensitive ◽  
Label Free Detection

The use of photonic crystals (PCS) in biosensor applications has lead to the development of highly sensitive and selective microfluidic sensor elements. Two main advantages of these devices for sensing applications are their high sensitivity and their reduced size, which makes it possible, in one hand, to detect very small analytes without the need of markers (label-free detection), and to integrate many of these devices on a single chip to perform a multi-parameter detection on the other hand. In the present paper, we analyze the design of a highly sensitive microfluidic sensors based on 2D photonic crystal slab waveguide formed by increasing the radii of air holes localized at each side of the line defect and filling with homogenous de-ionized water (nc =1.33). The transmission spectrum of the sensor has been obtained with the use of Finite Difference Time Domain (FDTD) method and it has been observed that a 306 nm wavelength position of the lower band edge shift was observed corresponding to a sensitivity of more than 927 nm per refractive index unit (RIU). Development of microfluidic sensor designs that enhance sensitivity is especially important because it allows detection of lower concentrations of analytes.

scholarly journals Label-Free Electrochemical Sensor for Rapid Bacterial Pathogen Detection Using Vancomycin-Modified Highly Branched Polymers

Sensors ◽  
2021 ◽  
Vol 21 (5) ◽  
pp. 1872
Author(s):  
Holger Schulze ◽  
Harry Wilson ◽  
Ines Cara ◽  
Steven Carter ◽  
Edward N. Dyson ◽  
...  
Keyword(s):  
Pathogen Detection ◽  
Electrochemical Impedance ◽  
Point Of Care ◽  
Branched Polymers ◽  
Label Free ◽  
Conformation Change ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
Staphylococcus Carnosus ◽  
Label Free Detection

Rapid point of care tests for bacterial infection diagnosis are of great importance to reduce the misuse of antibiotics and burden of antimicrobial resistance. Here, we have successfully combined a new class of non-biological binder molecules with electrochemical impedance spectroscopy (EIS)-based sensor detection for direct, label-free detection of Gram-positive bacteria making use of the specific coil-to-globule conformation change of the vancomycin-modified highly branched polymers immobilized on the surface of gold screen-printed electrodes upon binding to Gram-positive bacteria. Staphylococcus carnosus was detected after just 20 min incubation of the sample solution with the polymer-functionalized electrodes. The polymer conformation change was quantified with two simple 1 min EIS tests before and after incubation with the sample. Tests revealed a concentration dependent signal change within an OD600 range of Staphylococcus carnosus from 0.002 to 0.1 and a clear discrimination between Gram-positive Staphylococcus carnosus and Gram-negative Escherichia coli bacteria. This exhibits a clear advancement in terms of simplified test complexity compared to existing bacteria detection tests. In addition, the polymer-functionalized electrodes showed good storage and operational stability.

scholarly journals Highly Sensitive Fluorescent Detection of Acetylcholine Based on the Enhanced Peroxidase-Like Activity of Histidine Coated Magnetic Nanoparticles

Nanomaterials ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1207
Author(s):  
Hong Jae Cheon ◽  
Quynh Huong Nguyen ◽  
Moon Il Kim
Keyword(s):  
Magnetic Nanoparticles ◽  
High Sensitivity ◽  
High Specificity ◽  
Choline Oxidase ◽  
Fluorescent Detection ◽  
One Pot ◽  
Site Structure ◽  
Specificity And Sensitivity ◽  
Highly Sensitive ◽  
Peroxidase Mimics

Inspired by the active site structure of natural horseradish peroxidase having iron as a pivotal element with coordinated histidine residues, we have developed histidine coated magnetic nanoparticles (His@MNPs) with relatively uniform and small sizes (less than 10 nm) through one-pot heat treatment. In comparison to pristine MNPs and other amino acid coated MNPs, His@MNPs exhibited a considerably enhanced peroxidase-imitating activity, approaching 10-fold higher in catalytic reactions. With the high activity, His@MNPs then were exploited to detect the important neurotransmitter acetylcholine. By coupling choline oxidase and acetylcholine esterase with His@MNPs as peroxidase mimics, target choline and acetylcholine were successfully detected via fluorescent mode with high specificity and sensitivity with the limits of detection down to 200 and 100 nM, respectively. The diagnostic capability of the method is demonstrated by analyzing acetylcholine in human blood serum. This study thus demonstrates the potential of utilizing His@MNPs as peroxidase-mimicking nanozymes for detecting important biological and clinical targets with high sensitivity and reliability.

scholarly journals Microfluidic Impedance Biosensor Chips Using Sensing Layers Based on DNA-Based Self-Assembled Monolayers for Label-Free Detection of Proteins

Biosensors ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 80
Author(s):  
Khaled Alsabbagh ◽  
Tim Hornung ◽  
Achim Voigt ◽  
Sahba Sadir ◽  
Taleieh Rajabi ◽  
...  
Keyword(s):  
Troponin I ◽  
Electrochemical Impedance ◽  
Specific Binding ◽  
Self Assembled Monolayers ◽  
Significant Shift ◽  
Label Free ◽  
Label Free Detection ◽  
Assembled Monolayers ◽  
Self Assembled ◽  
Biosensor Chip

A microfluidic chip for electrochemical impedance spectroscopy (EIS) is presented as bio-sensor for label-free detection of proteins by using the example of cardiac troponin I. Troponin I is one of the most specific diagnostic serum biomarkers for myocardial infarction. The microfluidic impedance biosensor chip presented here consists of a microscope glass slide serving as base plate, sputtered electrodes, and a polydimethylsiloxane (PDMS) microchannel. Electrode functionalization protocols were developed considering a possible charge transfer through the sensing layer, in addition to analyte-specific binding by corresponding antibodies and reduction of nonspecific protein adsorption to prevent false-positive signals. Reagents tested for self-assembled monolayers (SAMs) on gold electrodes included thiolated hydrocarbons and thiolated oligonucleotides, where SAMs based on the latter showed a better performance. The corresponding antibody was covalently coupled on the SAM using carbodiimide chemistry. Sampling and measurement took only a few minutes. Application of a human serum albumin (HSA) sample, 1000 ng/mL, led to negligible impedance changes, while application of a troponin I sample, 1 ng/mL, led to a significant shift in the Nyquist plot. The results are promising regarding specific detection of clinically relevant concentrations of biomarkers, such as cardiac markers, with the newly developed microfluidic impedance biosensor chip.

Target-induced structure switching of DNA for label-free and ultrasensitive electrochemiluminescent detection of proteins

2014 ◽  
Vol 50 (24) ◽  
pp. 3211-3213 ◽  
Author(s):  
Mengli Yang ◽  
Ying Chen ◽  
Yun Xiang ◽  
Ruo Yuan ◽  
Yaqin Chai
Keyword(s):  
Dna Structure ◽  
Label Free ◽  
Switching Strategy ◽  
Highly Sensitive ◽  
Label Free Detection ◽  
Exo Iii ◽  
Recycling Amplification ◽  
Structure Switching ◽  
Induced Structure

Highly sensitive and label-free detection of thrombin is achieved via a target-induced DNA structure switching strategy and Exo III-assisted recycling amplification.

Photonic crystals: A platform for label-free and enhanced fluorescence biomolecular and cellular assays

2008 ◽  
Vol 1133 ◽  
Author(s):  
Brian T. Cunningham ◽  
Leo Chan ◽  
Patrick C. Mathias ◽  
Nikhil Ganesh ◽  
Sherine George ◽  
...  
Keyword(s):  
Photonic Crystal ◽  
Photonic Crystals ◽  
High Throughput Screening ◽  
High Sensitivity ◽  
Excitation Wavelength ◽  
Label Free ◽  
Crystal Surfaces ◽  
Enhanced Fluorescence ◽  
Preferred Direction ◽  
Label Free Detection

Abstract Photonic crystal surfaces represent a class of resonant optical structures that are capable of supporting high intensity electromagnetic standing waves with near-field and far-field properties that can be exploited for high sensitivity detection of biomolecules and cells. While modulation of the resonant wavelength of a photonic crystal by the dielectric permittivity of adsorbed biomaterials enables label-free detection, the resonance can also be tuned to coincide with the excitation wavelength of common fluorescent tags - including organic molecules and semiconductor quantum dots. Photonic crystals are also capable of efficiently channeling fluorescent emission into a preferred direction for enhanced extraction efficiency. Photonic crystals can be designed to support multiple resonant modes that can perform label free detection, enhanced fluorescence excitation, and enhanced fluorescence extraction simultaneously on the same device. Because photonic crystal surfaces may be inexpensively produced over large surface areas by nanoreplica molding processes, they can be incorporated into disposable labware for applications such as pharmaceutical high throughput screening. In this talk, the optical properties of surface photonic crystals will be reviewed and several applications will be described, including results from screening a 200,000-member chemical compound library for inhibitors of protein-DNA interactions, gene expression microarrays, and high sensitivity of protein biomarkers.

scholarly journals A Highly Sensitive Cell-Based TLR Reporter Platform for the Specific Detection of Bacterial TLR Ligands

2022 ◽  
Vol 12 ◽  
Author(s):  
Katharina Radakovics ◽  
Claire Battin ◽  
Judith Leitner ◽  
Sabine Geiselhart ◽  
Wolfgang Paster ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Ectopic Expression ◽  
High Sensitivity ◽  
High Specificity ◽  
Reporter System ◽  
Specificity And Sensitivity ◽  
Highly Sensitive ◽  
Definition Of ◽  
The Individual ◽  
Allergen Extracts

Toll-like receptors (TLRs) are primary pattern recognition receptors (PRRs), which recognize conserved microbial components. They play important roles in innate immunity but also in the initiation of adaptive immune responses. Impurities containing TLR ligands are a frequent problem in research but also for the production of therapeutics since TLR ligands can exert strong immunomodulatory properties even in minute amounts. Consequently, there is a need for sensitive tools to detect TLR ligands with high sensitivity and specificity. Here we describe the development of a platform based on a highly sensitive NF-κB::eGFP reporter Jurkat JE6-1 T cell line for the detection of TLR ligands. Ectopic expression of TLRs and their coreceptors and CRISPR/Cas9-mediated deletion of endogenously expressed TLRs was deployed to generate reporter cell lines selectively expressing functional human TLR2/1, TLR2/6, TLR4 or TLR5 complexes. Using well-defined agonists for the respective TLR complexes we could demonstrate high specificity and sensitivity of the individual reporter lines. The limit of detection for LPS was below 1 pg/mL and ligands for TLR2/1 (Pam3CSK4), TLR2/6 (Fsl-1) and TLR5 (flagellin) were detected at concentrations as low as 1.0 ng/mL, 0.2 ng/mL and 10 pg/mL, respectively. We showed that the JE6-1 TLR reporter cells have the utility to characterize different commercially available TLR ligands as well as more complex samples like bacterially expressed proteins or allergen extracts. Impurities in preparations of microbial compounds as well as the lack of specificity of detection systems can lead to erroneous results and currently there is no consensus regarding the involvement of TLRs in the recognition of several molecules with proposed immunostimulatory functions. This reporter system represents a highly suitable tool for the definition of structural requirements for agonists of distinct TLR complexes.

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