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High throughput microfluidics

Zhao Yu, Lu Zhou, Ting Zhang, Rui Shen, Chenxi Li, Xu Fang, Gareth Griffiths, Jian Liu
Matrix metalloproteinases (MMPs) are typically up-regulated in cancer cells, and play a critical role in assisting metastasis by the breakdown of the extracellular matrix. Here we report an effective strategy for cell invasiveness assay by integrating MMP9 functional activity analysis with single cell-encapsulated microdroplets. A flow focusing capillary microfluidic device has been assembled using "off-the-shelf" fluidic components for high-throughput generation of microdroplets. Tumor cells, MMP9 specific peptides, and other cofactors can be loaded into the device and encapsulated into individual droplets as dynamic microreactors for proteolytic cleavage of the substrate...
May 26, 2017: ACS Sensors
S Etcheverry, A Faridi, H Ramachandraiah, T Kumar, W Margulis, F Laurell, A Russom
Flow cytometry is currently the gold standard for analysis of cells in the medical laboratory and biomedical research. Fuelled by the need of point-of-care diagnosis, a significant effort has been made to miniaturize and reduce cost of flow cytometers. However, despite recent advances, current microsystems remain less versatile and much slower than their large-scale counterparts. In this work, an all-silica fibre microflow cytometer is presented that measures fluorescence and scattering from particles and cells...
July 17, 2017: Scientific Reports
Anson H L Tang, Queenie T K Lai, Bob M F Chung, Kelvin C M Lee, Aaron T Y Mok, G K Yip, Anderson H C Shum, Kenneth K Y Wong, Kevin K Tsia
Scaling the number of measurable parameters, which allows for multidimensional data analysis and thus higher-confidence statistical results, has been the main trend in the advanced development of flow cytometry. Notably, adding high-resolution imaging capabilities allows for the complex morphological analysis of cellular/sub-cellular structures. This is not possible with standard flow cytometers. However, it is valuable for advancing our knowledge of cellular functions and can benefit life science research, clinical diagnostics, and environmental monitoring...
June 28, 2017: Journal of Visualized Experiments: JoVE
Joshua Heinemann, Brigit Noon, Daniel Willems, Katherine Budeski, Brian Bothner
Elucidation and monitoring of biomarkers continues to expand because of their medical value and potential to reduce healthcare costs. For example, biomarkers are used extensively to track physiology associated with drug addiction, disease progression, aging, and industrial processes. While longitudinal analyses are of great value from a biological or healthcare perspective, the cost associated with replicate analyses is preventing the expansion of frequent routine testing. Frequent testing could deepen our understanding of disease emergence and aid adoption of personalized healthcare...
January 21, 2017: Analytical Methods: Advancing Methods and Applications
Xiaonan Yang, Zhuofa Chen, Jun Miao, Liwang Cui, Weihua Guan
This work reports a high throughput and label-free microfluidic cell deformability sensor for quantitative parasitemia measurement and stage determination for Plasmodium falciparum-infected red blood cells (Pf-iRBCs). The sensor relies on differentiating the RBC deformability (a mechanical biomarker) that is highly correlated with the infection status. The cell deformability is measured by evaluating the transit time when each individual RBC squeezes through a microscale constriction (cross-section ~5µm×5µm)...
July 8, 2017: Biosensors & Bioelectronics
Masahito Hosokawa, Yohei Nishikawa, Masato Kogawa, Haruko Takeyama
Massively parallel single-cell genome sequencing is required to further understand genetic diversities in complex biological systems. Whole genome amplification (WGA) is the first step for single-cell sequencing, but its throughput and accuracy are insufficient in conventional reaction platforms. Here, we introduce single droplet multiple displacement amplification (sd-MDA), a method that enables massively parallel amplification of single cell genomes while maintaining sequence accuracy and specificity. Tens of thousands of single cells are compartmentalized in millions of picoliter droplets and then subjected to lysis and WGA by passive droplet fusion in microfluidic channels...
July 12, 2017: Scientific Reports
Anastasia Ioanna Skilitsi, Timothé Turko, Damien Cianfarani, Sophie Barre, Wilfried Uhring, Ulrich Hassiepen, Jérémie Léonard
Time-resolved fluorescence detection for robust sensing of biomolecular interactions is developed by implementing Time Correlated Single Photon Counting in high-throughput conditions. Droplet microfluidics is used as a promising platform for the very fast handling of low-volume samples. We illustrate the potential of this very sensitive and cost-effective technology in the context of an enzymatic activity assay based on fluorescently-labeled biomolecules. Fluorescence lifetime detection by Time Correlated Single Photon Counting is shown to enable reliable discrimination between positive and negative control samples at a throughput as high as several hundred samples per second...
July 12, 2017: Methods and Applications in Fluorescence
Maira Shakeel Syed, Mehdi Rafeie, Rita Henderson, Dries Vandamme, Mohsen Asadnia, Majid Ebrahimi Warkiani
The separation of micro-sized particles in a continuous flow is crucial part of many industrial processes, from biopharmaceutical manufacturing to water treatment. Conventional separation techniques such as centrifugation and membrane filtration are largely limited by factors such as clogging, processing time and operation efficiency. Microfluidic based techniques have been gaining great attention in recent years as efficient and powerful approaches for particle-liquid separation. Yet the production of such systems using standard micro-fabrication techniques is proven to be tedious, costly and have cumbersome user interfaces, which all render commercialization difficult...
July 11, 2017: Lab on a Chip
Shinya Sakuma, Yusuke Kasai, Takeshi Hayakawa, Fumihito Arai
Although researchers have proposed various methods of on-chip cell sorting, high-throughput sorting of large cells remains hampered by the difficulty of controlling high-speed flow over a wide sorting area. To overcome this problem, we proposed high-speed local-flow control using dual membrane pumps driven by piezoelectric actuators placed on the outside of a microfluidic chip in this paper. We evaluated the controllability of shifting the flow profile by the local-flow. The results indicated that we could sort large cells up to approximately 150 μm in size with an equivalent throughput of 31 kHz...
July 7, 2017: Lab on a Chip
Jiang Xu, Shusheng Zhang, Anais Machado, Sébastien Lecommandoux, Olivier Sandre, Frank Gu, Annie Colin
We present a versatile continuous microfluidic flow-focusing method for the production of Doxorubicin (DOX) or Tamoxifen (TAM)-loaded poly(D,L-lactic-co-glycolic acid) (PLGA) nanoparticles (NPs). We use a partially water-miscible solvent mixture (dimethyl sulfoxide DMSO+ dichloromethane DCM) as precursor drug/polymer solution for NPs nucleation. We extrude this partially water-miscible solution into an aqueous medium and synthesized uniform PLGA NPs with higher drug loading ability and longer sustained-release ability than conventional microfluidic or batch preparation methods...
July 6, 2017: Scientific Reports
Brant Axt, Yi-Fan Hsieh, Divya Nalayanda, Tza-Huei Wang
Droplet microfluidics has found use in many biological assay applications as a means of high-throughput sample processing. One of the challenges of the technology, however, is the ability to control and merge droplets on-demand as they flow through the microdevices. It is in the interest of developing lab-on-chip devices to be able to combinatorically program additive mixing steps for more complex multistep and multiplex assays. Existing technologies to merge droplets are either passive in nature or require highly predictable droplet movement for feedforward control, making them vulnerable to errors during high throughput operation...
September 2017: Biomedical Microdevices
David N Carruthers, Chang Kyu Byun, Bradley J Cardinale, Xiaoxia Nina Lin
Algae are ubiquitous in natural ecosystems and have been studied extensively for biofuel production due to their unique metabolic capabilities. Most studies to date have approached biofuel optimization through synthetic biology and process engineering with few industrial scale projects considering algal community interactions. Such interactions can potentially lead to increased productivity and reduced community invasability, both important characteristics for scalable algal biofuel production. It is estimated that over a million species of algae exist such that elucidating the interactions that might be beneficial for biofuel production remains extremely resource and time intensive...
July 3, 2017: Integrative Biology: Quantitative Biosciences From Nano to Macro
Jie Zhang, Xiaofeng Wei, Rui Zeng, Feng Xu, XiuJun Li
Microfluidic lab-on-a-chip provides a new platform with unique advantages to mimic complex physiological microenvironments in vivo and has been increasingly exploited to stem cell research. In this review, we highlight recent advances of microfluidic devices for stem cell culture and differentiation toward the development of organ-on-a-chip, especially with an emphasis on vital innovations within the last 2 years. Various aspects for improving on-chip stem-cell culture and differentiation, particularly toward organ-on-a-chip, are discussed, along with microenvironment control, surface modification, extracellular scaffolds, high throughput and stimuli...
June 2017: Future Science OA
Michael M Soniat, Logan R Myler, Jeffrey M Schaub, Yoori Kim, Ignacio F Gallardo, Ilya J Finkelstein
Homologous recombination (HR) is a universally conserved DNA double-strand break repair pathway. Single-molecule fluorescence imaging approaches have revealed new mechanistic insights into nearly all aspects of HR. These methods are especially suited for studying protein complexes because multicolor fluorescent imaging can parse out subassemblies and transient intermediates that associate with the DNA substrates on the millisecond to hour timescales. However, acquiring single-molecule datasets remains challenging because most of these approaches are designed to measure one molecular reaction at a time...
2017: Methods in Enzymology
Rose J Brazilek, Francisco J Tovar-Lopez, Angus K T Wong, Huyen Tran, Amanda S Davis, James D McFadyen, Zane Kaplan, Sanjeev Chunilal, Shaun P Jackson, Harshal Nandurkar, Arnan Mitchell, Warwick S Nesbitt
Von Willebrand's disease (VWD) is the most common inherited bleeding disorder caused by either quantitative or qualitative defects of von Willebrand factor (VWF). Current tests for VWD require relatively large blood volumes, have low throughput, are time-consuming, and do not incorporate the physiologically relevant effects of haemodynamic forces. We developed a microfluidic device incorporating micro-contractions that harnesses well-defined haemodynamic strain gradients to initiate platelet aggregation in citrated whole blood...
June 29, 2017: Lab on a Chip
Yongmin Cho, Daniel A Porto, Hyundoo Hwang, Laura J Grundy, William R Schafer, Hang Lu
C. elegans is a useful genetic model system for investigating mechanisms involved in sensory behavior which are potentially relevant to human diseases. While utilities of advanced techniques such as microfluidics have accelerated some areas of C. elegans sensory biology such as chemosensation, studies of mechanosensation conventionally require immobilization by glue and manual delivery of stimuli, leading to low experimental throughput and high variability. Here we present a microfluidic platform that precisely and robustly delivers a wide range of mechanical stimuli and can also be used in conjunction with functional imaging and optical interrogation techniques...
June 29, 2017: Lab on a Chip
Ya Gai, Minkyu Kim, Ming Pan, Sindy K Y Tang
This paper describes the break-up behavior of a concentrated emulsion comprising drops stabilized by amphiphilic silica nanoparticles flowing in a tapered microchannel. Such geometry is often used in serial droplet interrogation and sorting processes in droplet microfluidics applications. When exposed to high viscous stresses, drops can undergo break-up and compromise their physical integrity. As these drops are used as micro-reactors, such compromise leads to a loss in the accuracy of droplet-based assays...
May 2017: Biomicrofluidics
Lucas R Blauch, Ya Gai, Jian Wei Khor, Pranidhi Sood, Wallace F Marshall, Sindy K Y Tang
Wound repair is a key feature distinguishing living from nonliving matter. Single cells are increasingly recognized to be capable of healing wounds. The lack of reproducible, high-throughput wounding methods has hindered single-cell wound repair studies. This work describes a microfluidic guillotine for bisecting single Stentor coeruleus cells in a continuous-flow manner. Stentor is used as a model due to its robust repair capacity and the ability to perform gene knockdown in a high-throughput manner. Local cutting dynamics reveals two regimes under which cells are bisected, one at low viscous stress where cells are cut with small membrane ruptures and high viability and one at high viscous stress where cells are cut with extended membrane ruptures and decreased viability...
June 26, 2017: Proceedings of the National Academy of Sciences of the United States of America
Paola Occhetta, Roberta Visone, Marco Rasponi
The design of innovative tools for generating physiologically relevant three-dimensional (3D) in vitro models has been recently recognized as a fundamental step to study cell responses and long-term tissue functionalities thanks to its ability to recapitulate the complexity and the dimensional scale of the cellular microenvironment, while directly integrating high-throughput and automatic screening capabilities.This chapter addresses the development of a poly(dimethylsiloxane)-based microfluidic platform to (1) generate and culture 3D cellular microaggregates under continuous flow perfusion while (2) conditioning them with different combinations/concentrations of soluble factors (i...
2017: Methods in Molecular Biology
Wenming Liu, Jinyi Wang
Three-dimensional (3D) tumor culture miniaturized platforms are of importance to biomimetic model construction and pathophysiological studies. Controllable and high-throughput production of 3D tumors is desirable to make cell-based manipulation dynamic and efficient at micro-scale. Moreover, the 3D culture platform being reusable is convenient to research scholars. In this chapter, we describe a dynamically controlled 3D tumor manipulation and culture method using pneumatic microstructure-based microfluidics, which has potential applications in the fields of tissue engineering, tumor biology, and clinical medicine in a high-throughput way...
2017: Methods in Molecular Biology
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