scholarly journals High-Throughput and Label-Free Single Nanoparticle Sizing Based on Time-Resolved On-Chip Microscopy

ACS Nano ◽  
2015 ◽  
Vol 9 (3) ◽  
pp. 3265-3273 ◽  
Author(s):  
Euan McLeod ◽  
T. Umut Dincer ◽  
Muhammed Veli ◽  
Yavuz N. Ertas ◽  
Chau Nguyen ◽  
...  
Keyword(s):  
High Throughput ◽  
Label Free ◽  
Time Resolved ◽  
Single Nanoparticle ◽  
On Chip

High-Throughput and Label-Free Detection of Motile Parasites in Bodily Fluids Using Lensless Time-Resolved Speckle Imaging

2019 ◽  
Author(s):  
Yibo Zhang ◽  
Hatice Ceylan Koydemir ◽  
Michelle M. Shimogawa ◽  
Sener Yalcin ◽  
Alexander Guziak ◽  
...  
Keyword(s):  
High Throughput ◽  
Label Free ◽  
Speckle Imaging ◽  
Time Resolved ◽  
Bodily Fluids ◽  
Label Free Detection

Monitoring On-Chip Pictet-Spengler Reactions by Integrated Analytical Separation and Label-Free Time-Resolved Fluorescence

2011 ◽  
Vol 18 (4) ◽  
pp. 1240-1246 ◽  
Author(s):  
Stefan Ohla ◽  
Reinhild Beyreiss ◽  
Stefanie Fritzsche ◽  
Petra Glaser ◽  
Stefan Nagl ◽  
...  
Keyword(s):  
Free Time ◽  
Label Free ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
On Chip ◽  
Analytical Separation

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.

scholarly journals Lipid-Sensing High-Throughput ApoA-I Assays

2012 ◽  
Vol 17 (8) ◽  
pp. 1050-1061 ◽  
Author(s):  
Anita Niedziela-Majka ◽  
Latesh Lad ◽  
Jeffrey W. Chisholm ◽  
Leanna Lagpacan ◽  
Karen Schwartz ◽  
...  
Keyword(s):  
High Throughput ◽  
Dynamic Range ◽  
Cholesterol Metabolism ◽  
Resonance Energy Transfer ◽  
Density Lipoprotein ◽  
Resonance Energy ◽  
Wavelength Shift ◽  
Label Free ◽  
Time Resolved ◽  
Increased Risk

Apolipoprotein A-I (ApoA-I), a primary protein component of high-density lipoprotein (HDL), plays an important role in cholesterol metabolism mediating the formation of HDL and the efflux of cellular cholesterol from macrophage foam cells in arterial walls. Lipidation of ApoA-I is mediated by adenosine triphosphate (ATP) binding cassette A1 (ABCA1). Insufficient ABCA1 activity may lead to increased risk of atherosclerosis due to reduced HDL formation and cholesterol efflux. The standard radioactive assay for measuring cholesterol transport to ApoA-I has low throughput and poor dynamic range, and it fails to measure phospholipid transfer. We describe the development of two sensitive, nonradioactive high-throughput assays that report on the lipidation of ApoA-I: a homogeneous assay based on time-resolved fluorescence resonance energy transfer (TR-FRET) and a discontinuous assay that uses the label-free Epic platform. The TR-FRET assay employs HiLyte Fluor 647–labeled ApoA-I with N-terminal biotin bound to streptavidin-terbium. When fluorescent ApoA-I was incorporated into HDL, TR-FRET decreased proportionally to the increase in the ratio of lipids to ApoA-I, demonstrating that the assay was sensitive to the amount of lipid bound to ApoA-I. In the Epic assay, biotinylated ApoA-I was captured on a streptavidin-coated biosensor. Measured resonant wavelength shift was proportional to the amount of lipids associated with ApoA-I, indicating that the assay senses ApoA-I lipidation.

scholarly journals Use of High-Throughput Mass Spectrometry to Reduce False Positives in Protease uHTS Screens

2014 ◽  
Vol 20 (2) ◽  
pp. 212-222 ◽  
Author(s):  
Gregory C. Adam ◽  
Juncai Meng ◽  
Joseph M. Rizzo ◽  
Adam Amoss ◽  
Jeffrey W. Lusen ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
High Throughput Screening ◽  
Fluorescent Dyes ◽  
False Positives ◽  
Indirect Detection ◽  
Detection Methods ◽  
Label Free ◽  
Fluorescent Compound ◽  
Time Resolved

As a label-free technology, mass spectrometry (MS) enables assays to be generated that monitor the conversion of substrates with native sequences to products without the requirement for substrate modifications or indirect detection methods. Although traditional liquid chromatography (LC)–MS methods are relatively slow for a high-throughput screening (HTS) paradigm, with cycle times typically ≥60 s per sample, the Agilent RapidFire High-Throughput Mass Spectrometry (HTMS) System, with a cycle time of 5–7 s per sample, enables rapid analysis of compound numbers compatible with HTS. By monitoring changes in mass directly, HTMS assays can be used as a triaging tool by eliminating large numbers of false positives resulting from fluorescent compound interference or from compounds interacting with hydrophobic fluorescent dyes appended to substrates. Herein, HTMS assays were developed for multiple protease programs, including cysteine, serine, and aspartyl proteases, and applied as a confirmatory assay. The confirmation rate for each protease assay averaged <30%, independent of the primary assay technology used (i.e., luminescent, fluorescent, and time-resolved fluorescent technologies). Importantly, >99% of compounds designed to inhibit the enzymes were confirmed by the corresponding HTMS assay. Hence, HTMS is an effective tool for removing detection-based false positives from ultrahigh-throughput screening, resulting in hit lists enriched in true actives for downstream dose response titrations and hit-to-lead efforts.

Time-Resolved Luminescent High-Throughput Screening Platform for Lysosomotropic Compounds in Living Cells

ACS Sensors ◽  
2020 ◽  
Author(s):  
Ke-Jia Wu ◽  
Chun Wu ◽  
Feng Chen ◽  
Sha-Sha Cheng ◽  
Dik-Lung Ma ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Living Cells ◽  
Time Resolved ◽  
Screening Platform

scholarly journals High‐throughput dilution‐based growth method enables time‐resolved exo‐metabolomics of Pseudomonas putida and Pseudomonas aeruginosa

2021 ◽  
Author(s):  
Bjarke H. Pedersen ◽  
Nicolás Gurdo ◽  
Helle Krogh Johansen ◽  
Søren Molin ◽  
Pablo I. Nikel ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Pseudomonas Putida ◽  
High Throughput ◽  
Time Resolved ◽  
Growth Method

Single‐Nanoparticle Coulometry Method with High Sensitivity and High Throughput to Study the Electrochemical Activity and Oscillation of Single Nanocatalysts

Small ◽  
2021 ◽  
Vol 17 (14) ◽  
pp. 2007302
Author(s):  
Mohan Lin ◽  
Yingke Zhou ◽  
Lingzheng Bu ◽  
Chuang Bai ◽  
Muhammad Tariq ◽  
...  
Keyword(s):  
High Throughput ◽  
High Sensitivity ◽  
Electrochemical Activity ◽  
Single Nanoparticle

scholarly journals Development of a Robust High-Throughput Screening Platform for Inhibitors of the Striatal-Enriched Tyrosine Phosphatase (STEP)

2021 ◽  
Vol 22 (9) ◽  
pp. 4417
Author(s):  
Lester J Lambert ◽  
Stefan Grotegut ◽  
Maria Celeridad ◽  
Palak Gosalia ◽  
Laurent JS De Backer ◽  
...  
Keyword(s):  
Drug Discovery ◽  
High Throughput ◽  
Tyrosine Kinases ◽  
High Throughput Screening ◽  
Kinase Inhibitors ◽  
Tyrosine Phosphatase ◽  
Protein Tyrosine Phosphatases ◽  
Synaptic Function ◽  
Label Free ◽  
Protein Tyrosine

Many human diseases are the result of abnormal expression or activation of protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs). Not surprisingly, more than 30 tyrosine kinase inhibitors (TKIs) are currently in clinical use and provide unique treatment options for many patients. PTPs on the other hand have long been regarded as “undruggable” and only recently have gained increased attention in drug discovery. Striatal-enriched tyrosine phosphatase (STEP) is a neuron-specific PTP that is overactive in Alzheimer’s disease (AD) and other neurodegenerative and neuropsychiatric disorders, including Parkinson’s disease, schizophrenia, and fragile X syndrome. An emergent model suggests that the increase in STEP activity interferes with synaptic function and contributes to the characteristic cognitive and behavioral deficits present in these diseases. Prior efforts to generate STEP inhibitors with properties that warrant clinical development have largely failed. To identify novel STEP inhibitor scaffolds, we developed a biophysical, label-free high-throughput screening (HTS) platform based on the protein thermal shift (PTS) technology. In contrast to conventional HTS using STEP enzymatic assays, we found the PTS platform highly robust and capable of identifying true hits with confirmed STEP inhibitory activity and selectivity. This new platform promises to greatly advance STEP drug discovery and should be applicable to other PTP targets.

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