scholarly journals High-Throughput Cell Trapping in the Dentate Spiral Microfluidic Channel

Micromachines ◽  
2021 ◽  
Vol 12 (3) ◽  
pp. 288
Author(s):  
Jiawei Lu ◽  
Bo Dai ◽  
Kan Wang ◽  
Yan Long ◽  
Zhuoqing Yang ◽  
...  

Cell trapping is a very useful technique in a variety of cell-based assays and cellular research fields. It requires a high-throughput, high-efficiency operation to isolate cells of interest and immobilize the captured cells at specific positions. In this study, a dentate spiral microfluidic structure is proposed for cell trapping. The structure consists of a main spiral channel connecting an inlet and an out and a large number of dentate traps on the side of the channel. The density of the traps is high. When a cell comes across an empty trap, the cell suddenly makes a turn and enters the trap. Once the trap captures enough cells, the trap becomes closed and the following cells pass by the trap. The microfluidic structure is optimized based on the investigation of the influence over the flow. In the demonstration, 4T1 mouse breast cancer cells injected into the chip can be efficiently captured and isolated in the different traps. The cell trapping operates at a very high flow rate (40 μL/s) and a high trapping efficiency (>90%) can be achieved. The proposed high-throughput cell-trapping technique can be adopted in the many applications, including rapid microfluidic cell-based assays and isolation of rare circulating tumor cells from a large volume of blood sample.

Author(s):  
Hayato Nishizaki ◽  
Taisuke Fukuda ◽  
Kyohei Terao ◽  
Hidekuni Takao ◽  
Fumikazu Oohira ◽  
...  

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
R. Brearton ◽  
L. A. Turnbull ◽  
J. A. T. Verezhak ◽  
G. Balakrishnan ◽  
P. D. Hatton ◽  
...  

AbstractMagnetic skyrmions are topologically non-trivial, swirling magnetization textures that form lattices in helimagnetic materials. These magnetic nanoparticles show promise as high efficiency next-generation information carriers, with dynamics that are governed by their topology. Among the many unusual properties of skyrmions is the tendency of their direction of motion to deviate from that of a driving force; the angle by which they diverge is a materials constant, known as the skyrmion Hall angle. In magnetic multilayer systems, where skyrmions often appear individually, not arranging themselves in a lattice, this deflection angle can be easily measured by tracing the real space motion of individual skyrmions. Here we describe a reciprocal space technique which can be used to determine the skyrmion Hall angle in the skyrmion lattice state, leveraging the properties of the skyrmion lattice under a shear drive. We demonstrate this procedure to yield a quantitative measurement of the skyrmion Hall angle in the room-temperature skyrmion system FeGe, shearing the skyrmion lattice with the magnetic field gradient generated by a single turn Oersted wire.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Andrew T. Meek ◽  
Nils M. Kronenberg ◽  
Andrew Morton ◽  
Philipp Liehm ◽  
Jan Murawski ◽  
...  

AbstractImportant dynamic processes in mechanobiology remain elusive due to a lack of tools to image the small cellular forces at play with sufficient speed and throughput. Here, we introduce a fast, interference-based force imaging method that uses the illumination of an elastic deformable microcavity with two rapidly alternating wavelengths to map forces. We show real-time acquisition and processing of data, obtain images of mechanical activity while scanning across a cell culture, and investigate sub-second fluctuations of the piconewton forces exerted by macrophage podosomes. We also demonstrate force imaging of beating neonatal cardiomyocytes at 100 fps which reveals mechanical aspects of spontaneous oscillatory contraction waves in between the main contraction cycles. These examples illustrate the wider potential of our technique for monitoring cellular forces with high throughput and excellent temporal resolution.


2016 ◽  
Vol 22 (S3) ◽  
pp. 1080-1081
Author(s):  
Alberto Estevez ◽  
Colin Garvey ◽  
Claudio Ciferri

2008 ◽  
Vol 78 (2) ◽  
pp. A48
Author(s):  
William Severson ◽  
Joseph Maddry ◽  
Xi Chen ◽  
Subramaniam Ananthan ◽  
Adrian Poffenberger ◽  
...  

2015 ◽  
Vol 124 ◽  
pp. 20-29 ◽  
Author(s):  
Zongyi Hu ◽  
Xin Hu ◽  
Shanshan He ◽  
Hyung Joon Yim ◽  
Jingbo Xiao ◽  
...  

2016 ◽  
Vol 22 (1) ◽  
pp. 77-85 ◽  
Author(s):  
Aleksandra R. Dukic ◽  
David W. McClymont ◽  
Kjetil Taskén

Connexin 43 (Cx43), the predominant gap junction (GJ) protein, directly interacts with the A-kinase-anchoring protein (AKAP) Ezrin in human cytotrophoblasts and a rat liver epithelial cells (IAR20). The Cx43-Ezrin–protein kinase (PKA) complex facilitates Cx43 phosphorylation by PKA, which triggers GJ opening in cytotrophoblasts and IAR20 cells and may be a general mechanism regulating GJ intercellular communication (GJIC). Considering the importance of Cx43 GJs in health and disease, they are considered potential pharmaceutical targets. The Cx43-Ezrin interaction is a protein-protein interaction that opens possibilities for targeting with peptides and small molecules. For this reason, we developed a high-throughput cell-based assay in which GJIC can be assessed and new compounds characterized. We used two pools of IAR20 cells, calcein loaded and unloaded, that were mixed and allowed to attach. Next, GJIC was monitored over time using automated imaging via the IncuCyte imager. The assay was validated using known GJ inhibitors and anchoring peptide disruptors, and we further tested new peptides that interfered with the Cx43-Ezrin binding region and reduced GJIC. Although an AlphaScreen assay can be used to screen for Cx43-Ezrin interaction inhibitors, the cell-based assay described is an ideal secondary screen for promising small-molecule hits to help identify the most potent compounds.


2018 ◽  
Vol 19 (11) ◽  
pp. 3569 ◽  
Author(s):  
Lilas Courtot ◽  
Jean-Sébastien Hoffmann ◽  
Valérie Bergoglio

Genome stability requires tight regulation of DNA replication to ensure that the entire genome of the cell is duplicated once and only once per cell cycle. In mammalian cells, origin activation is controlled in space and time by a cell-specific and robust program called replication timing. About 100,000 potential replication origins form on the chromatin in the gap 1 (G1) phase but only 20–30% of them are active during the DNA replication of a given cell in the synthesis (S) phase. When the progress of replication forks is slowed by exogenous or endogenous impediments, the cell must activate some of the inactive or “dormant” origins to complete replication on time. Thus, the many origins that may be activated are probably key to protect the genome against replication stress. This review aims to discuss the role of these dormant origins as safeguards of the human genome during replicative stress.


2011 ◽  
Vol 16 (8) ◽  
pp. 925-931 ◽  
Author(s):  
Amy Emery ◽  
David A. Sorrell ◽  
Stacy Lawrence ◽  
Emma Easthope ◽  
Mark Stockdale ◽  
...  

Aurora A kinase is a key regulator of mitosis, which is upregulated in several human cancers, making it a potential target for anticancer therapeutics. Consequently, robust medium- to high-throughput cell-based assays to measure Aurora A kinase activity are critical for the development of small-molecule inhibitors. Here the authors compare measurement of the phosphorylation of two Aurora A substrates previously used in high-content screening Aurora A assays, Aurora A itself and TACC3, with a novel substrate Lats2. Using antibodies directed against phosphorylated forms of Aurora A (pThr288), P-TACC3 (pSer558), and P-Lats2 (pSer83), the authors investigate their suitability in parallel for development of a cell-based assay using several reference Aurora inhibitors: MLN8054, VX680, and AZD1152-HQPA. They validate a combined assay of target-specific phosphorylation of Lats2 at the centrosome and an increase in mitotic index as a measure of Aurora A activity. The assay is both sensitive and robust and has acceptable assay performance for high-throughput screening or potency estimation from concentration–response assays. It has the advantage that it can be carried out using a commercially available monoclonal antibody against phospho-Lats2 and the widely available Cellomics ArrayScan HCS reader and thus represents a significant addition to the tools available for the identification of Aurora A specific inhibitors.


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