scholarly journals A high-throughput label-free cell-based biosensor (CBB) system

2010 ◽  
Author(s):  
Feng Xu ◽  
SangJun Moon ◽  
Evan Hefner ◽  
Turker Beyazoglu ◽  
Ahmet E. Emre ◽  
...  
Keyword(s):  
High Throughput ◽  
Free Cell ◽  
Label Free

scholarly journals Multi-angle pulse shape detection of scattered light in flow cytometry for label-free cell cycle classification

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Daniel Kage ◽  
Kerstin Heinrich ◽  
Konrad v. Volkmann ◽  
Jenny Kirsch ◽  
Kristen Feher ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
High Throughput ◽  
Pulse Shape ◽  
Single Cells ◽  
Scattered Light ◽  
Free Cell ◽  
Hek293 Cells ◽  
Label Free ◽  
Functional Changes

AbstractFlow cytometers are robust and ubiquitous tools of biomedical research, as they enable high-throughput fluorescence-based multi-parametric analysis and sorting of single cells. However, analysis is often constrained by the availability of detection reagents or functional changes of cells caused by fluorescent staining. Here, we introduce MAPS-FC (multi-angle pulse shape flow cytometry), an approach that measures angle- and time-resolved scattered light for high-throughput cell characterization to circumvent the constraints of conventional flow cytometry. In order to derive cell-specific properties from the acquired pulse shapes, we developed a data analysis procedure based on wavelet transform and k-means clustering. We analyzed cell cycle stages of Jurkat and HEK293 cells by MAPS-FC and were able to assign cells to the G1, S, and G2/M phases without the need for fluorescent labeling. The results were validated by DNA staining and by sorting and re-analysis of isolated G1, S, and G2/M populations. Our results demonstrate that MAPS-FC can be used to determine cell properties that are otherwise only accessible by invasive labeling. This approach is technically compatible with conventional flow cytometers and paves the way for label-free cell sorting.

scholarly journals Label-free cell cycle analysis for high-throughput imaging flow cytometry

2016 ◽  
Vol 7 (1) ◽  
Author(s):  
Thomas Blasi ◽  
Holger Hennig ◽  
Huw D. Summers ◽  
Fabian J. Theis ◽  
Joana Cerveira ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
High Throughput ◽  
Free Cell ◽  
Cell Cycle Analysis ◽  
Label Free ◽  
Imaging Flow Cytometry ◽  
High Throughput Imaging

Label-free cell phenotypic study of opioid receptors and discovery of novel mu opioid ligands from natural products

2021 ◽  
Vol 270 ◽  
pp. 113872
Author(s):  
Tao Hou ◽  
Fangfang Xu ◽  
Xingrong Peng ◽  
Han Zhou ◽  
Xiuli Zhang ◽  
...  
Keyword(s):  
Natural Products ◽  
Opioid Receptors ◽  
Free Cell ◽  
Label Free ◽  
Mu Opioid ◽  
Opioid Ligands

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.

scholarly journals Development of a Novel Label-Free and High-Throughput Arginase-1 Assay Using Self-Assembled Monolayer Desorption Ionization Mass Spectrometry

2021 ◽  
pp. 247255522110006
Author(s):  
Michael D. Scholle ◽  
Zachary A. Gurard-Levin
Keyword(s):  
Mass Spectrometry ◽  
Drug Discovery ◽  
High Throughput ◽  
Ionization Mass Spectrometry ◽  
Ionization Mass ◽  
Label Free ◽  
Desorption Ionization ◽  
Arginase 1 ◽  
Self Assembled ◽  
High Throughput Assay

Arginase-1, an enzyme that catalyzes the reaction of L-arginine to L-ornithine, is implicated in the tumor immune response and represents an interesting therapeutic target in immuno-oncology. Initiating arginase drug discovery efforts remains a challenge due to a lack of suitable high-throughput assay methodologies. This report describes the combination of self-assembled monolayers and matrix-assisted laser desorption ionization mass spectrometry to enable the first label-free and high-throughput assay for arginase activity. The assay was optimized for kinetically balanced conditions and miniaturized, while achieving a robust assay (Z-factor > 0.8) and a significant assay window [signal-to-background ratio > 20] relative to fluorescent approaches. To validate the assay, the inhibition of the reference compound nor-NOHA (Nω-hydroxy-nor-L-arginine) was evaluated, and the IC50 measured to be in line with reported results (IC50 = 180 nM). The assay was then used to complete a screen of 175,000 compounds, demonstrating the high-throughput capacity of the approach. The label-free format also eliminates opportunities for false-positive results due to interference from library compounds and optical readouts. The assay methodology described here enables new opportunities for drug discovery for arginase and, due to the assay flexibility, can be more broadly applicable for measuring other amino acid–metabolizing enzymes.

Clog-free cell filtration using resettable cell traps

Lab on a Chip ◽  
2014 ◽  
Vol 14 (15) ◽  
pp. 2657-2665 ◽  
Author(s):  
William Beattie ◽  
Xi Qin ◽  
Lin Wang ◽  
Hongshen Ma
Keyword(s):  
High Throughput ◽  
Cell Separation ◽  
Free Cell ◽  
Separation Mechanism ◽  
Cell Filtration ◽  
Capture And Release

A microfluidic cell separation mechanism created using constrictions with adjustable size that can selectively capture and release cells, thereby enabling high throughput size and deformability based cell separation without clogging.

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