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High throughput microfluidics

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https://www.readbyqxmd.com/read/28520400/femtomole-scale-high-throughput-screening-of-protein-ligands-with-droplet-based-thermal-shift-assay
#1
Wen-Wen Liu, Ying Zhu, Qun Fang
There is a great demand to measure protein-ligand interactions in rapid and low cost way. Here we developed a microfluidic droplet-based thermal shift assay (dTSA) system for high throughput screening of small-molecule protein ligands. The system is composed of a nanoliter droplet array chip, a microfluidic droplet robot, and a real-time fluorescence detection system. Total 324 assays could be performed in parallel in a single chip with an 18 × 18 droplet array. The consumption of dTSA for each protein or ligand sample was only 5 nL (femtomole scale), which is significantly reduced by over 3 orders of magnitude compared with those in 96 or 384-well plate-based systems...
May 18, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28515430/a-microfluidic-chip-for-screening-individual-cancer-cells-via-eavesdropping-on-autophagy-inducing-crosstalk-in-the-stroma-niche
#2
Hacer Ezgi Karakas, Junyoung Kim, Juhee Park, Jung Min Oh, Yongjun Choi, Devrim Gozuacik, Yoon-Kyoung Cho
Autophagy is a cellular homeostatic mechanism where proteins and organelles are digested and recycled to provide an alternative source of building blocks and energy to cells. The role of autophagy in cancer microenvironment is still poorly understood. Here, we present a microfluidic system allowing monitoring of the crosstalk between single cells. We used this system to study how tumor cells induced autophagy in the stromal niche. Firstly, we could confirm that transforming growth factor β1 (TGFβ1) secreted from breast tumor cells is a paracrine mediator of tumor-stroma interaction leading to the activation of autophagy in the stroma component fibroblasts...
May 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28513735/highly-efficient-genome-editing-of-human-hematopoietic-stem-cells-via-a-nano-silicon-blade-delivery-approach
#3
Yuan Ma, Xin Han, Oscar Quintana Bustamante, Ricardo Bessa de Castro, Kai Zhang, Pengchao Zhang, Ying Li, Zongbin Liu, Xuewu Liu, Mauro Ferrari, Zhongbo Hu, José Carlos Segovia, Lidong Qin
Recently, the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 bacterial immunity system has opened a promising avenue to treat genetic diseases that affect the human hematopoietic stem cells (HSCs). Therefore, finding a highly efficient delivery method capable of modifying the genome in the hard-to-transfect HSCs, combined with the advanced CRISPR-Cas9 system, may meet the challenges for dissecting the hematologic disease mechanisms and facilitate future clinical applications. Here, we developed an effective HSC-specified delivery microfluidic chip to disrupt the cell membrane transiently by inducing rapid mechanical deformation that allowed the delivery of biomaterials into the cytoplasm from the surrounding matrix...
May 17, 2017: Integrative Biology: Quantitative Biosciences From Nano to Macro
https://www.readbyqxmd.com/read/28512313/a-rapid-capillary-pressure-driven-micro-channel-to-demonstrate-newtonian-fluid-behavior-of-zebrafish-blood-at-high-shear-rates
#4
Juhyun Lee, Tzu-Chieh Chou, Dongyang Kang, Hanul Kang, Junjie Chen, Kyung In Baek, Wei Wang, Yichen Ding, Dino Di Carlo, Yu-Chong Tai, Tzung K Hsiai
Blood viscosity provides the rheological basis to elucidate shear stress underlying developmental cardiac mechanics and physiology. Zebrafish is a high throughput model for developmental biology, forward-genetics, and drug discovery. The micro-scale posed an experimental challenge to measure blood viscosity. To address this challenge, a microfluidic viscometer driven by surface tension was developed to reduce the sample volume required (3μL) for rapid (<2 min) and continuous viscosity measurement. By fitting the power-law fluid model to the travel distance of blood through the micro-channel as a function of time and channel configuration, the experimentally acquired blood viscosity was compared with a vacuum-driven capillary viscometer at high shear rates (>500 s(-1)), at which the power law exponent (n) of zebrafish blood was nearly 1 behaving as a Newtonian fluid...
May 16, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28509918/generation-and-manipulation-of-hydrogel-microcapsules-by-droplet-based-microfluidics-for-mammalian-cell-culture
#5
REVIEW
Haishui Huang, Yin Yu, Yong Hu, Xiaoming He, O Berk Usta, Martin L Yarmush
Hydrogel microcapsules provide miniaturized and biocompatible niches for three-dimensional (3D) in vitro cell culture. They can be easily generated by droplet-based microfluidics with tunable size, morphology, and biochemical properties. Therefore, microfluidic generation and manipulation of cell-laden microcapsules can be used for 3D cell culture to mimic the in vivo environment towards applications in tissue engineering and high throughput drug screening. In this review of recent advances mainly since 2010, we will first introduce general characteristics of droplet-based microfluidic devices for cell encapsulation with an emphasis on the fluid dynamics of droplet breakup and internal mixing as they directly influence microcapsule's size and structure...
May 16, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/28509913/optimization-of-a-polydopamine-pd-based-coating-method-and-polydimethylsiloxane-pdms-substrates-for-improved-mouse-embryonic-stem-cell-esc-pluripotency-maintenance-and-cardiac-differentiation
#6
Jiayin Fu, Yon Jin Chuah, Wee Tong Ang, Nan Zheng, Dong-An Wang
Myocardiocyte derived from pluripotent stem cells, such as induced pluripotent stem cells (iPSCs) and embryonic stem cells (ESCs), is a promising cell source for cardiac tissue engineering. Combined with microfluidic technologies, a heart-on-a-chip is very likely to be developed and function as a platform for high throughput drug screening. Polydimethylsiloxane (PDMS) silicone elastomer is a widely-used biomaterial for the investigation of cell-substrate interactions and biochip fabrication. However, the intrinsic PDMS surface hydrophobicity inhibits cell adhesion on the PDMS surface, and PDMS surface modification is required for effective cell adhesion...
May 16, 2017: Biomaterials Science
https://www.readbyqxmd.com/read/28492663/a-versatile-microfluidic-device-for-high-throughput-production-of-microparticles-and-cell-microencapsulation
#7
Samin Akbari, Tohid Pirbodaghi, Roger D Kamm, Paula T Hammond
Biocompatible microparticles are valuable tools in biomedical research for applications such as drug delivery, cell transplantation therapy, and analytical assays. However, their translation into clinical research and the pharmaceutical industry has been slow due to the lack of techniques that can produce microparticles with controlled physicochemical properties at high throughput. We introduce a robust microfluidic platform for the production of relatively homogeneous microdroplets at a generation frequency of up to 3...
May 11, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/28481895/succinct-workflows-for-circulating-tumor-cells-after-enrichment-from-systematic-counting-to-mutational-profiling
#8
Victor Chun-Lam Wong, Josephine Mun-Yee Ko, Chi-Tat Lam, Maria Li Lung
PURPOSE: This study aims to establish a highly adaptable workflow downstream of microfluidic enrichment for facilitating systematic CTC enumeration and genetic discovery. METHODS: To facilitate CTC enumeration, we established a CK/EPCAM-combined immunostaining strategy and an automated CTC analytical pipeline using an open-source image analyzer. By virtue of this workflow, we conducted a pilot study of 56 cancer patients and 21 healthy individuals using a high-throughput spiral microfluidic chip system...
2017: PloS One
https://www.readbyqxmd.com/read/28481437/effective-light-directed-assembly-of-building-blocks-with-microscale-control
#9
Ngoc-Duy Dinh, Rongcong Luo, Maria Tankeh Asuncion Christine, Weikang Nicholas Lin, Wei-Chuan Shih, James Cho-Hong Goh, Chia-Hung Chen
Light-directed forces have been widely used to pattern micro/nanoscale objects with precise control, forming functional assemblies. However, a substantial laser intensity is required to generate sufficient optical gradient forces to move a small object in a certain direction, causing limited throughput for applications. A high-throughput light-directed assembly is demonstrated as a printing technology by introducing gold nanorods to induce thermal convection flows that move microparticles (diameter = 40 µm to several hundreds of micrometers) to specific light-guided locations, forming desired patterns...
May 8, 2017: Small
https://www.readbyqxmd.com/read/28469762/x-ray-transparent-microfluidic-chips-for-high-throughput-screening-and-optimization-of-in-meso-membrane-protein-crystallization
#10
Jeremy M Schieferstein, Ashtamurthy S Pawate, Chang Sun, Frank Wan, Paige N Sheraden, Jana Broecker, Oliver P Ernst, Robert B Gennis, Paul J A Kenis
Elucidating and clarifying the function of membrane proteins ultimately requires atomic resolution structures as determined most commonly by X-ray crystallography. Many high impact membrane protein structures have resulted from advanced techniques such as in meso crystallization that present technical difficulties for the set-up and scale-out of high-throughput crystallization experiments. In prior work, we designed a novel, low-throughput X-ray transparent microfluidic device that automated the mixing of protein and lipid by diffusion for in meso crystallization trials...
March 2017: Biomicrofluidics
https://www.readbyqxmd.com/read/28467352/screening-genotoxicity-chemistry-with-microfluidic-electrochemiluminescent-arrays
#11
REVIEW
Itti Bist, Kiran Bano, James F Rusling
This review describes progress in the development of electrochemiluminescent (ECL) arrays aimed at sensing DNA damage to identify genotoxic chemistry related to reactive metabolites. Genotoxicity refers to chemical or photochemical processes that damage DNA with toxic consequences. Our arrays feature DNA/enzyme films that form reactive metabolites of test chemicals that can subsequently react with DNA, thus enabling prediction of genotoxic chemical reactions. These high-throughput ECL arrays incorporating representative cohorts of human metabolic enzymes provide a platform for determining chemical toxicity profiles of new drug and environmental chemical candidates...
May 3, 2017: Sensors
https://www.readbyqxmd.com/read/28467055/microfluidic-technology-uncovering-the-mechanisms-of-nanocrystal-nucleation-and-growth
#12
Ioannis Lignos, Richard Maceiczyk, Andrew J deMello
The controlled and reproducible formation of colloidal semiconductor nanocrystals (or quantum dots) is of central importance in nanoscale science and technology. The tunable size- and shape-dependent properties of such materials make them ideal candidates for the development of efficient and low-cost displays, solar cells, light-emitting devices, and catalysts. The formidable difficulties associated with the macroscale preparation of semiconductor nanocrystals (possessing bespoke optical and chemical properties) result from the fact that underlying reaction mechanisms are complex and that the reactive environment is difficult to control...
May 3, 2017: Accounts of Chemical Research
https://www.readbyqxmd.com/read/28466284/a-microfluidic-diode-for-sorting-and-immobilization-of-caenorhabditis-elegans
#13
Lijie Yang, Tao Hong, Yin Zhang, Jose Guillermo Sanchez Arriola, Brian L Nelms, Richard Mu, Deyu Li
Caenorhabditis elegans (C. elegans) is a powerful model organism extensively used in studies of human aging and diseases. Despite the numerous advantages of C. elegans as a model system, two biological characteristics may introduce complexity and variability to most studies: 1. it exhibits different biological features, composition and behaviors at different developmental stages; 2. it has very high mobility. Therefore, synchronization and immobilization of worm populations are often required. Conventionally, these processes are implemented through manual and chemical methods, which can be laborious, time-consuming and of low-throughput...
June 2017: Biomedical Microdevices
https://www.readbyqxmd.com/read/28465615/high-throughput-single-cell-counting-in-droplet-based-microfluidics
#14
Heng Lu, Ouriel Caen, Jeremy Vrignon, Eleonora Zonta, Zakaria El Harrak, Philippe Nizard, Jean-Christophe Baret, Valérie Taly
Droplet-based microfluidics is extensively and increasingly used for high-throughput single-cell studies. However, the accuracy of the cell counting method directly impacts the robustness of such studies. We describe here a simple and precise method to accurately count a large number of adherent and non-adherent human cells as well as bacteria. Our microfluidic hemocytometer provides statistically relevant data on large populations of cells at a high-throughput, used to characterize cell encapsulation and cell viability during incubation in droplets...
May 2, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28462964/synthesis-of-new-hydrophilic-rhodamine-based-enzymatic-substrates-compatible-with-droplet-based-microfluidic-assays
#15
Johan Fenneteau, Dany Chauvin, Andrew D Griffiths, Clément Nizak, Janine Cossy
Here we report the conception, synthesis and evaluation of new hydrophilic rhodamine-based enzymatic substrates for detection of peptidase activity compatible with high-throughput screening using droplet-based microfluidics.
May 11, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28448040/determination-of-high-affinity-antibody-antigen-binding-kinetics-using-four-biosensor-platforms
#16
Danlin Yang, Ajit Singh, Helen Wu, Rachel Kroe-Barrett
Label-free optical biosensors are powerful tools in drug discovery for the characterization of biomolecular interactions. In this study, we describe the use of four routinely used biosensor platforms in our laboratory to evaluate the binding affinity and kinetics of ten high-affinity monoclonal antibodies (mAbs) against human proprotein convertase subtilisin kexin type 9 (PCSK9). While both Biacore T100 and ProteOn XPR36 are derived from the well-established Surface Plasmon Resonance (SPR) technology, the former has four flow cells connected by serial flow configuration, whereas the latter presents 36 reaction spots in parallel through an improvised 6 x 6 crisscross microfluidic channel configuration...
April 17, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28448036/using-microfluidic-devices-to-measure-lifespan-and-cellular-phenotypes-in-single-budding-yeast-cells
#17
Ke Zou, Diana S Ren, Qi Ou-Yang, Hao Li, Jiashun Zheng
Budding yeast Saccharomyces cerevisiae is an important model organism in aging research. Genetic studies have revealed many genes with conserved effects on the lifespan across species. However, the molecular causes of aging and death remain elusive. To gain a systematic understanding of the molecular mechanisms underlying yeast aging, we need high-throughput methods to measure lifespan and to quantify various cellular and molecular phenotypes in single cells. Previously, we developed microfluidic devices to track budding yeast mother cells throughout their lifespan while flushing away newborn daughter cells...
March 30, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28447031/microfluidic-concentric-gradient-generator-design-for-high-throughput-cell-based-studies
#18
Elishai Ezra Tsur, Michal Zimerman, Idan Maor, Avner Elrich, Yaakov Nahmias
Gradients of diffusible signaling molecules play important role in various processes, ranging from cell differentiation to toxicological evaluation. Microfluidic technology provides an accurate control of tempospatial conditions. However, current microfluidic platforms are not designed to handle multiple gradients and cell populations simultaneously. Here, we demonstrate a rapidly adaptable microfluidic design able to expose multiple cell populations to an array of chemical gradients. Our design is based on pressure-equilibrated concentric channels and a pressure-dissipating control layer, facilitating the seeding of multiple cell populations in a single device...
2017: Frontiers in Bioengineering and Biotechnology
https://www.readbyqxmd.com/read/28446009/applications-of-microfluidic-devices-for-urology
#19
REVIEW
Se Jik Han, Hun-Kuk Park, Kyung Sook Kim
Microfluidics is considered an important technology that is suitable for numerous biomedical applications, including cancer diagnosis, metastasis, drug delivery, and tissue engineering. Although microfluidics is still considered to be a new approach in urological research, several pioneering studies have been reported in recent years. In this paper, we reviewed urological research works using microfluidic devices. Microfluidic devices were used for the detection of prostate and bladder cancer and the characterization of cancer microenvironments...
April 2017: International Neurourology Journal
https://www.readbyqxmd.com/read/28445605/high-throughput-cell-mechanical-phenotyping-for-label-free-titration-assays-of-cytoskeletal-modifications
#20
Stefan Golfier, Philipp Rosendahl, Alexander Mietke, Maik Herbig, Jochen Guck, Oliver Otto
The mechanical fingerprint of cells is inherently linked to the structure of the cytoskeleton and can serve as a label free marker for cell homeostasis or pathologic states. How cytoskeletal composition affects the physical response of cells to external loads has been intensively studied with a spectrum of techniques, yet quantitative and statistically powerful investigations in the form of titration assays are hampered by the low throughput of most available methods. In this study, we employ real-time deformability cytometry (RT-DC), a novel microfluidic tool to examine the effects of biochemically modified F-actin and microtubule stability and nuclear chromatin structure on cell deformation in a human leukemia cell line (HL60)...
April 26, 2017: Cytoskeleton
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