scholarly journals Use of LSPR Spectroscopy Biosensing for In Situ Identification of Arsenic from Bioleaching of Realgar by Acidithiobacillus ferrooxidans

2018 ◽  
Vol 2018 ◽  
pp. 1-6
Author(s):  
Ruixiang Xu ◽  
Wenbin Zhao ◽  
Shenghui Xu ◽  
Lei Yan ◽  
Zhengrong Wu ◽  
...  
Keyword(s):  
High Throughput ◽  
Low Cost ◽  
Response Speed ◽  
Localized Surface Plasmon ◽  
Enzyme Linked Immunosorbent Assay ◽  
Arsenic Content ◽  
Label Free ◽  
Monitoring Methods ◽  
Specific Selectivity

Localized surface plasmon resonance (LSPR) spectroscopy has received extensive attention as a new method in chemical and biological analysis that can be integrated with nanotechnology. As a new analytical method, LSPR possesses various advantages over the traditional bioanalysis method of enzyme-linked immunosorbent assay (ELISA), including a label-free procedure, low cost, high response speed, simple operation and structure, and ease of storage and transport. Additionally, in situ and high-throughput measurements can be achieved. This study aims to solve the problem of the lack of in situ and highly efficient monitoring methods for current realgar bioleaching processes. An LSPR chip is made to monitor the changes in arsenic content in the process of realgar bioleaching. A convenient, fast, sensitive, and high-throughput method of bioleaching process investigation based on the LSPR spectroscopic in situ monitoring technique is proposed. The LSPR chip provided a highly specific selectivity and a linear detection of arsenic content in the range of 1.0–100.0 μM with detection limit (LOD) 0.898 μM. The developed chip was applied to the quantification of realgar bioleaching sample with satisfactory results.

scholarly journals Integration of an In Situ MALDI-Based High-Throughput Screening Process: A Case Study with Receptor Tyrosine Kinase c-MET

2017 ◽  
Vol 22 (10) ◽  
pp. 1203-1210 ◽  
Author(s):  
Katrin Beeman ◽  
Jens Baumgärtner ◽  
Manuel Laubenheimer ◽  
Karlheinz Hergesell ◽  
Martin Hoffmann ◽  
...  
Keyword(s):  
Drug Discovery ◽  
High Throughput ◽  
High Throughput Screening ◽  
Maldi Ms ◽  
Kinase Assay ◽  
Label Free ◽  
Screening Process ◽  
Label Free Detection ◽  
Ic50 Values

Mass spectrometry (MS) is known for its label-free detection of substrates and products from a variety of enzyme reactions. Recent hardware improvements have increased interest in the use of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS for high-throughput drug discovery. Despite interest in this technology, several challenges remain and must be overcome before MALDI-MS can be integrated as an automated “in-line reader” for high-throughput drug discovery. Two such hurdles include in situ sample processing and deposition, as well as integration of MALDI-MS for enzymatic screening assays that usually contain high levels of MS-incompatible components. Here we adapt our c-MET kinase assay to optimize for MALDI-MS compatibility and test its feasibility for compound screening. The pros and cons of the Echo (Labcyte) as a transfer system for in situ MALDI-MS sample preparation are discussed. We demonstrate that this method generates robust data in a 1536-grid format. We use the MALDI-MS to directly measure the ratio of c-MET substrate and phosphorylated product to acquire IC50 curves and demonstrate that the pharmacology is unaffected. The resulting IC50 values correlate well between the common label-based capillary electrophoresis and the label-free MALDI-MS detection method. We predict that label-free MALDI-MS-based high-throughput screening will become increasingly important and more widely used for drug discovery.

scholarly journals MALDIViz: A Comprehensive Informatics Tool for MALDI-MS Data Visualization and Analysis

2017 ◽  
Vol 22 (10) ◽  
pp. 1246-1252 ◽  
Author(s):  
Kishore Kumar Jagadeesan ◽  
Simon Ekström
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
High Throughput Screening ◽  
Web Application ◽  
Low Cost ◽  
Maldi Ms ◽  
Ionization Mass ◽  
Label Free ◽  
Label Free Detection ◽  
R Shiny

Recently, mass spectrometry (MS) has emerged as an important tool for high-throughput screening (HTS) providing a direct and label-free detection method, complementing traditional fluorescent and colorimetric methodologies. Among the various MS techniques used for HTS, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) provides many of the characteristics required for high-throughput analyses, such as low cost, speed, and automation. However, visualization and analysis of the large datasets generated by HTS MALDI-MS can pose significant challenges, especially for multiparametric experiments. The datasets can be generated fast, and the complexity of the experimental data (e.g., screening many different sorbent phases, the sorbent mass, and the load, wash, and elution conditions) makes manual data analysis difficult. To address these challenges, a comprehensive informatics tool called MALDIViz was developed. This tool is an R-Shiny-based web application, accessible independently of the operating system and without the need to install any program locally. It has been designed to facilitate easy analysis and visualization of MALDI-MS datasets, comparison of multiplex experiments, and export of the analysis results to high-quality images.

Single Cell Interrogation using Optofluidic Platforms for Systems Immunology

MRS Advances ◽  
2016 ◽  
Vol 1 (56) ◽  
pp. 3783-3788 ◽  
Author(s):  
Serap Aksu
Keyword(s):  
Immune System ◽  
Single Cell ◽  
High Throughput ◽  
Individual Cell ◽  
Cytokine Secretion ◽  
Time Dependent ◽  
Label Free ◽  
System Behavior ◽  
Systems Immunology

ABSTRACTThe main objective of this report is to demonstrate novel engineering technologies to investigate regulatory mechanisms of systems immunology in a time-dependent and high-throughput manner. Understanding of immune system behavior is crucial for accurate prognosis of infections and identification of diseases at early stage. An ultimate goal of biomedical engineering is to develop predictive models of immune system behavior in tissue, which necessitates a comprehensive map of dynamic (time-dependent) input-output relationships at the individual cell level. Traditionally, biochemical analysis on the cell signaling is obtained from bulky cell ensembles which average over relevant individual cell response. The response consists firstly of signaling protein (cytokine) secretions which are released during a disease state and which are used to activate the immune system to respond to the disease. We investigate the cytokine secretion dynamics of a single immune cell in response to the stimulant using automated and comprehensive optofluidic platforms. These platforms enable survival and manipulation of single cells in compartments having compatible sizes with cells as well as provide precise control over the type, dose and time-course of the stimulant. The cytokine secretion dynamics of single cell are typically explained by measuring the types, rates, frequencies and concentrations of various cytokines. For the quantitative measurements, label free localized surface plasmon resonance (LSPR) based biosensor can be integrated within the microfluidic device. Microfluidic channels can confine secreted cytokines in compartments, minimize dilution effects and increase detection sensitivity for label free plasmonic biosensing. The direct application of LSPR to in-situ live cell function analysis is still in its infancy and use of such in-situ, real time, and label free biodetection will effortlessly provide high-throughput quantitative bioanalysis for understanding immune system behavior.

Ordered Au Nanodisk and Nanohole Arrays: Fabrication and Applications

2010 ◽  
Vol 1 (3) ◽  
Author(s):  
Yue Bing Zheng ◽  
Bala Krishna Juluri ◽  
Brian Kiraly ◽  
Tony Jun Huang
Keyword(s):  
Surface Plasmon Resonance ◽  
High Throughput ◽  
Localized Surface Plasmon Resonance ◽  
Low Cost ◽  
Localized Surface Plasmon ◽  
Dependent Behavior ◽  
Nanohole Arrays ◽  
Time Observation ◽  
Nanofabrication Technique ◽  
Real Time Observation

We have utilized nanosphere lithography (NSL) to fabricate ordered Au nanodisk and nanohole arrays on substrates and have studied the localized surface plasmon resonance (LSPR) of the arrays. Through these investigations, we demonstrate that the angle-dependent behavior of the LSPR in the Au nanodisk arrays enables real-time observation of exciton-plasmon couplings. In addition, we show that the NSL-fabricated Au nanohole arrays can be applied as templates for patterning micro-/nanoparticles under capillary force. The unique structural and plasmonic characteristics of the Au nanodisk and nanohole arrays, as well as the low-cost and high-throughput NSL-based nanofabrication technique, render these arrays excellent platforms for numerous engineering applications.

scholarly journals High-throughput patterning of photonic structures with tunable periodicity

2015 ◽  
Vol 112 (17) ◽  
pp. 5309-5313 ◽  
Author(s):  
Thomas J. Kempa ◽  
D. Kwabena Bediako ◽  
Sun-Kyung Kim ◽  
Hong-Gyu Park ◽  
Daniel G. Nocera
Keyword(s):  
Cyclic Voltammetry ◽  
High Throughput ◽  
Low Cost ◽  
Optical Elements ◽  
Periodic Arrays ◽  
Sensing Applications ◽  
Scan Rate ◽  
Reactive Interface

A patterning method termed “RIPPLE” (reactive interface patterning promoted by lithographic electrochemistry) is applied to the fabrication of arrays of dielectric and metallic optical elements. This method uses cyclic voltammetry to impart patterns onto the working electrode of a standard three-electrode electrochemical setup. Using this technique and a template stripping process, periodic arrays of Ag circular Bragg gratings are patterned in a high-throughput fashion over large substrate areas. By varying the scan rate of the cyclically applied voltage ramps, the periodicity of the gratings can be tuned in situ over micrometer and submicrometer length scales. Characterization of the periodic arrays of periodic gratings identified point-like and annular scattering modes at different planes above the structured surface. Facile, reliable, and rapid patterning techniques like RIPPLE may enable the high-throughput and low-cost fabrication of photonic elements and metasurfaces for energy conversion and sensing applications.

scholarly journals Label-Free Detection of Chondroitin Sulphate Proteoglycan 4 by a Polyaniline/Graphene Nanocomposite Functionalized Impedimetric Immunosensor

2016 ◽  
Vol 2016 ◽  
pp. 1-8 ◽  
Author(s):  
JingJing Fu ◽  
ZhuanZhuan Shi ◽  
Man Li ◽  
Yangyang Wang ◽  
Ling Yu
Keyword(s):  
Cell Lines ◽  
Chondroitin Sulphate ◽  
Low Cost ◽  
Enzyme Linked Immunosorbent Assay ◽  
Label Free ◽  
Sulphate Proteoglycan ◽  
Chondroitin Sulphate Proteoglycan ◽  
Label Free Detection ◽  
Proteoglycan 4 ◽  
Impedimetric Immunosensor

The chondroitin sulphate proteoglycan 4 (CSPG4), also known as high molecular weight-melanoma associated antigen (HMW-MAA), is a tumor-associated antigen that is expressed in more than 85% of surgically removed melanoma lesions but has restricted distribution in normal tissues. The diagnostic and therapeutic value of CSPG4 drives a need for sensitive and low-cost detection approaches. To this end, we developed a polyaniline/graphene oxide nanocomposite (PANI@GO) that was electrochemically codeposited on indium tin oxide (ITO) electrode. Glutaraldehyde mediated the covalent immobilization of CSPG4 specific antibody mAbD2.8.5 to construct a CSPG4 immunosensor using cell culture media and cell lysate as samples. The fully assembled impedimetric immunosensor was used to detect CSPG4 in CSPG4-positive cell lines M14/CSPG4 and MV3. No impedance signal changes could be observed from CSPG4-negative cell lines M14 and mAbMk2-23 showing the specificity of the CSPG4-impedimetric immunosensor. This low-cost, simple, and label-free analytical method is an alternative to enzyme-linked immunosorbent assay and flow cytometry in screening of CSPG4 in complex biological samples.

scholarly journals Rescue and In Situ Selection and Evaluation (RISE): A Method for High-Throughput Panning of Phage Display Libraries

2005 ◽  
Vol 10 (2) ◽  
pp. 108-117 ◽  
Author(s):  
Thomas Vanhercke ◽  
Christophe Ampe ◽  
Luc Tirry ◽  
Peter Denolf
Keyword(s):  
Phage Display ◽  
High Throughput ◽  
High Throughput Screening ◽  
Enzyme Linked Immunosorbent Assay ◽  
Selective Enrichment ◽  
Phage Display Libraries ◽  
Viral Particles ◽  
Phage Libraries ◽  
Rapid Characterization

Phage display has proven to be an invaluable instrument in the search for proteins and peptides with optimized or novel functions. The amplification and selection of phage libraries typically involve several operations and handling large bacterial cultures during each round. Purification of the assembled phage particles after rescue adds to the labor and time demand. The authors therefore devised a method, termed rescue and in situ selection and evaluation (RISE), which combines all steps from rescue to binding in a single microwell. To test this concept, wells were precoated with different antibodies, which allowed newly formed phage particles to be captured directly in situ during overnight rescue. Following 6 washing steps, the retained phages could be easily detected in an enzyme-linked immunosorbent assay (ELISA), thus eliminating the need for purification or concentration of the viral particles. As a consequence, RISE enables a rapid characterization of phage-displayed proteins. In addition, this method allowed for the selective enrichment of phages displaying a hemagglutinin (HA) epitope tag, spiked in a 104-fold excess of wild-type background. Because the combination of phage rescue, selection, or evaluation in a single microwell is amenable to automation, RISE may boost the high-throughput screening of smaller sized phage display libraries.

scholarly journals DISCA: high-throughput cryo-ET structural pattern mining by deep unsupervised clustering

2021 ◽  
Author(s):  
Xiangrui Zeng ◽  
Anson Kahng ◽  
Liang Xue ◽  
Julia Mahamid ◽  
Yi-Wei Chang ◽  
...  
Keyword(s):  
High Throughput ◽  
Pattern Mining ◽  
Electron Tomography ◽  
Structural Features ◽  
Computer Assisted ◽  
Label Free ◽  
Structural Pattern ◽  
Cryo Electron Tomography ◽  
Homogeneous Structures

Cryo-electron tomography directly visualizes heterogeneous macromolecular structures in complex cellular environments, but existing computer-assisted sorting approaches are low-throughput or inherently limited due to their dependency on available templates and manual labels. We introduce a high-throughput template-and-label-free deep learning approach that automatically discovers subsets of homogeneous structures by learning and modeling 3D structural features and their distributions. Diverse structures emerging from sorted subsets enable systematic unbiased recognition of macromolecular complexes in situ.

scholarly journals Plasmonic Nanopores: Optofluidic Separation of Nano-Bioparticles via Negative Depletion

2021 ◽  
Author(s):  
Xiangchao Zhu ◽  
Ahmet Cicek ◽  
Yixiang Li ◽  
Ahmet Ali Yanik
Keyword(s):  
Refractive Index ◽  
Chemical Composition ◽  
High Throughput ◽  
Light Transmission ◽  
Low Cost ◽  
Light Sources ◽  
Label Free ◽  
Incoherent Light ◽  
Drag Forces ◽  
On Chip

In this chapter, we review a novel “optofluidic” nanopore device enabling label-free sorting of nano-bioparticles [e.g., exosomes, viruses] based-on size or chemical composition. By employing a broadband objective-free light focusing mechanism through extraordinary light transmission effect, our plasmonic nanopore device eliminates sophisticated instrumentation requirements for precise alignment of optical scattering and fluidic drag forces, a fundamental shortcoming of the conventional optical chromatography techniques. Using concurrent optical gradient and radial fluidic drag forces, it achieves self-collimation of nano-bioparticles with inherently minimized spatial dispersion against the fluidic flow. This scheme enables size-based fractionation through negative depletion and refractive-index based separation of nano-bioparticles from similar size particles that have different chemical composition. Most remarkably, its small (4 μm × 4 μm) footprint facilitates on-chip, multiplexed, high-throughput nano-bioparticle sorting using low-cost incoherent light sources.

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