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Martyna Adamowicz, Klio Maratou, Timothy J Aitman
Whole genome shotgun bisulfite sequencing is a method used to generate genome-wide methylation profiles. There are many available protocols to validate the results of this genome-wide method, but they mostly share the limitation of measuring methylation at a small number of CpG positions in small numbers of samples. We developed a multiplexed DNA methylation analysis protocol, which allows for the simultaneous quantitative measurement of cytosine methylation at single nucleotide resolution in 48 PCR amplicons and 48 samples utilizing the microfluidic system established by Fluidigm...
2018: Methods in Molecular Biology
Reza Kowsari-Esfahan, Azadeh Jahanbakhsh, Mohammad Said Saidi, Shahin Bonakdar
Microfluidic devices are beneficial in miniaturizing and multiplexing various cellular assays in a single platform. Chondrogenesis is known to pertain to chemical, topographical, and mechanical cues in the microenvironment. Mechanical cues themselves have numerous parameters such as strain magnitude, frequency, and stimulation time. Effects of different strain magnitudes on the chondrogenic differentiation of adult stem cells have not been explored thoroughly. Here, a new multilayer microdevice is presented for the unidirectional compressive stimulation of cells in a three-dimensional cell culture...
December 5, 2017: Journal of the Mechanical Behavior of Biomedical Materials
J Nagashima, R El Assal, N Songsasen, U Demirci
The ability to grow oocytes from immature ovarian follicles in vitro has significant potential for fertility preservation; yet, it has proved challenging in large mammalian species due to the complex metabolic needs and long-term culture requirements. Currently, follicular incubations are based on a 'static' system with manual exchange of medium. Despite the numerous advantages of conventional culturing approaches, recapitulating the native microenvironment and supporting the survival of ovarian follicles still represent challenges...
December 8, 2017: Journal of Tissue Engineering and Regenerative Medicine
Alessio Lugnan, Joni Dambre, Peter Bienstman
The computational power required to classify cell holograms is a major limit to the throughput of label-free cell sorting based on digital holographic microscopy. In this work, a simple integrated photonic stage comprising a collection of silica pillar scatterers is proposed as an effective nonlinear mixing interface between the light scattered by a cell and an image sensor. The light processing provided by the photonic stage allows for the use of a simple linear classifier implemented in the electric domain and applied on a limited number of pixels...
November 27, 2017: Optics Express
Jiyun Jeong, Yeolin Lee, Yeongeun Yoo, Myung Kyu Lee
Agarose gel can be used for three dimensional (3D) cell culture because it prevents cell attachment. The dried agarose film coated on a culture plate also protected cell attachment and allowed 3D growth of cancer cells. We developed an efficient method for agarose film coating on an oxygen-plasma treated micropost polystyrene chip prepared by an injection molding process. The agarose film was modified to maleimide or Ni-NTA groups for covalent or cleavable attachment of photoactivatable Fc-specific antibody binding proteins (PFcBPs) via their N-terminal cysteine residues or 6xHis tag, respectively...
December 5, 2017: Colloids and Surfaces. B, Biointerfaces
Peng Hu, Emily Fabyanic, Deborah Y Kwon, Sheng Tang, Zhaolan Zhou, Hao Wu
Massively parallel single-cell RNA sequencing can precisely resolve cellular diversity in a high-throughput manner at low cost, but unbiased isolation of intact single cells from complex tissues such as adult mammalian brains is challenging. Here, we integrate sucrose-gradient-assisted purification of nuclei with droplet microfluidics to develop a highly scalable single-nucleus RNA-seq approach (sNucDrop-seq), which is free of enzymatic dissociation and nucleus sorting. By profiling ∼18,000 nuclei isolated from cortical tissues of adult mice, we demonstrate that sNucDrop-seq not only accurately reveals neuronal and non-neuronal subtype composition with high sensitivity but also enables in-depth analysis of transient transcriptional states driven by neuronal activity, at single-cell resolution, in vivo...
December 7, 2017: Molecular Cell
Flavia Vitale, Daniel Vercosa, Alexander V Rodriguez, Sushma Sri Pamulapati, Frederik Seibt, Eric Lewis, Jiaxi Stephen Yan, Krishna Badhiwala, Mohammed Adnan, Gianni Royer-Carfagni, Michael Beierlein, Caleb Kemere, Matteo Pasquali, Jacob Robinson
Soft and conductive nanomaterials like carbon nanotubes, graphene, and nanowire scaffolds have expanded the family of ultra-flexible microelectrodes that can bend and flex with the natural movement of the brain, reduce the inflammatory response and improve the stability of long-term neural recordings. However, current methods to implant these highly flexible electrodes rely on temporary stiffening agents that temporarily increase the electrode size and stiffness thus aggravating neural damage during implantation, which can lead to cell loss and glial activation that persists even after the stiffening agents are removed or dissolve...
December 8, 2017: Nano Letters
Xudong Zhu, Ju Chu, Yonghong Wang
The field of microbiology has traditionally been concerned with and focused on studies at the population level. Microfluidic platforms have emerged as important tools for biology research at a small scale, even down to a single cell level. The spatial and temporal control of cells and stimuli transported by microfluidic channels in well-designed microsystems realized the studies of specific cells in a controlled microenvironment. The true cellular physiology responses, which were obtained mostly by inference from population-level data, could be revealed in this way...
December 8, 2017: Biotechnology Journal
Jinsoo Park, Jin Ho Jung, Kwangseok Park, Ghulam Destgeer, Husnain Ahmed, Raheel Ahmad, Hyung Jin Sung
On-chip droplet splitting is one of the fundamental droplet-based microfluidic unit operations to control droplet volume after production and increase operational capability, flexibility, and throughput. Various droplet splitting methods have been proposed, and among them the acoustic droplet splitting method is promising because of its label-free operation without any physical or thermal damage to droplets. Previous acoustic droplet splitting methods faced several limitations: first, they employed a cross-type acoustofluidic device that precluded multichannel droplet splitting; second, they required irreversible bonding between a piezoelectric substrate and a microfluidic chip, such that the fluidic chip was not replaceable...
December 8, 2017: Lab on a Chip
Joshua Ricouvier, Romain Pierrat, Rémi Carminati, Patrick Tabeling, Pavel Yazhgur
We study long range density fluctuations (hyperuniformity) in two-dimensional jammed packings of bidisperse droplets. Taking advantage of microfluidics, we systematically span a large range of size and concentration ratios of the two droplet populations. We identify various defects increasing long range density fluctuations mainly due to organization of local particle environment. By choosing an appropriate bidispersity, we fabricate materials with a high level of hyperuniformity. Interesting transparency properties of these optimized materials are established based on numerical simulations...
November 17, 2017: Physical Review Letters
Céleste Odier, Bertrand Levaché, Enric Santanach-Carreras, Denis Bartolo
We establish a comprehensive description of the patterns formed when a wetting liquid displaces a viscous fluid confined in a porous medium. Building on model microfluidic experiments, we evidence four imbibition scenarios all yielding different large-scale morphologies. Combining high-resolution imaging and confocal microscopy, we show that they originate from two liquid-entrainment transitions and a Rayleigh-Plateau instability at the pore scale. Finally, we demonstrate and explain the long-time coarsening of the resulting patterns...
November 17, 2017: Physical Review Letters
Niek Welkenhuysen, Caroline B Adiels, Mattias Goksör, Stefan Hohmann
Microfluidic systems in combination with microscopy (e.g., fluorescence) can be a powerful tool to study, at single-cell level, the behavior and morphology of biological cells after uptake of glucose. Here, we briefly discuss the advantages of using microfluidic systems. We further describe how microfluidic systems are fabricated and how they are utilized. Finally, we discuss how the large amount of data can be analyzed in a "semi-automatic" manner using custom-made software. In summary, we provide a guide to how to use microfluidic systems in single-cell studies...
2018: Methods in Molecular Biology
Carlos Busatto, Juan Pesoa, Ignacio Helbling, Julio Luna, Diana Estenoz
Poly(lactic-co-glycolic acid) (PLGA) microparticles containing progesterone were prepared by the solvent extraction/evaporation and microfluidic techniques. Microparticles were characterized by their size distribution, encapsulation efficiency, morphology and thermal properties. The effect of particle size, polydispersity and polymer degradation on the in vitro release of the hormone was studied. A triphasic release profile was observed for larger microparticles, while smaller microspheres showed a biphasic release profile...
December 4, 2017: International Journal of Pharmaceutics
Iván Hernández-Neuta, Iago Pereiro, Annika Ahlford, Davide Ferraro, Qiongdi Zhang, Jean-Louis Viovy, Stéphanie Descroix, Mats Nilsson
Magnetic solid phase substrates for biomolecule manipulation have become a valuable tool for simplification and automation of molecular biology protocols. However, the handling of magnetic particles inside microfluidic chips for miniaturized assays is often challenging due to inefficient mixing, aggregation, and the advanced instrumentation required for effective actuation. Here, we describe the use of a microfluidic magnetic fluidized bed approach that enables dynamic, highly efficient and simplified magnetic bead actuation for DNA analysis in a continuous flow platform with minimal technical requirements...
November 26, 2017: Biosensors & Bioelectronics
Kyung Nim Noh, Sung Il Park, Raza Qazi, Zhanan Zou, Aaron D Mickle, Jose G Grajales-Reyes, Kyung-In Jang, Robert W Gereau, Jianliang Xiao, John A Rogers, Jae-Woong Jeong
Combination of optogenetics and pharmacology represents a unique approach to dissect neural circuitry with high specificity and versatility. However, conventional tools available to perform these experiments, such as optical fibers and metal cannula, are limited due to their tethered operation and lack of biomechanical compatibility. To address these issues, a miniaturized, battery-free, soft optofluidic system that can provide wireless drug delivery and optical stimulation for spatiotemporal control of the targeted neural circuit in freely behaving animals is reported...
December 7, 2017: Small
Bee Luan Khoo, Gianluca Grenci, Ying Bena Lim, Soo Chin Lee, Jongyoon Han, Chwee Teck Lim
The development of personalized cancer therapy depends on a robust system to monitor the patient's individual response to anticancer treatment. Anticancer drug efficacy has been tested on circulating tumor cells (CTCs) derived from patient blood samples after ex vivo expansion into CTC clusters. Current attempts to culture these primary cancer cells focus on long-term maintenance under growth factor supplements into cell lines, which usually takes >6 months and results in a CTC expansion efficiency of <20%...
January 2018: Nature Protocols
Anke Urbansky, Pelle Ohlsson, Andreas Lenshof, Fabio Garofalo, Stefan Scheding, Thomas Laurell
Effective separation methods for fractionating blood components are needed for numerous diagnostic and research applications. This paper presents the use of acoustophoresis, an ultrasound based microfluidic separation technology, for label-free, gentle and continuous separation of mononuclear cells (MNCs) from diluted whole blood. Red blood cells (RBCs) and MNCs behave similar in an acoustic standing wave field, compromising acoustic separation of MNC from RBC in standard buffer systems. However, by optimizing the buffer conditions and thereby changing the acoustophoretic mobility of the cells, we were able to enrich MNCs relative to RBCs by a factor of 2,800 with MNC recoveries up to 88%...
December 7, 2017: Scientific Reports
Lucie A Low, Danilo A Tagle
The scientific and technological development of microphysiological systems (MPS) modeling organs-on-chips, or "tissue chips" (TCs), has progressed rapidly over the past decade. Stem cell research and microfluidic concepts have combined to lead to the development of microphysiological platforms representing an ever-expanding list of different human organ systems. In the context of rare diseases, these bioengineered microfluidics platforms hold promise for modeling of disorders and could prove useful in the screening and efficacy testing of existing therapeutics...
2017: Advances in Experimental Medicine and Biology
Xiangpeng Li, Christopher J Easley
Recent breakthroughs in organ-on-a-chip and related technologies have highlighted the extraordinary potential for microfluidics to not only make lasting impacts in the understanding of biological systems but also to create new and important in vitro culture platforms. Adipose tissue (fat), in particular, is one that should be amenable to microfluidic mimics of its microenvironment. While the tissue was traditionally considered important only for energy storage, it is now understood to be an integral part of the endocrine system that secretes hormones and responds to various stimuli...
December 6, 2017: Analytical and Bioanalytical Chemistry
I Rodríguez-Ruiz, V Babenko, S Martínez-Rodríguez, J A Gavira
Lab-on-a-Chip (LoC), or micro-Total Analysis Systems (μTAS), is recognized as a powerful analytical technology with high capabilities, though end-user products for protein purification are still far from being available on the market. Remarkable progress has been achieved in the separation of nucleic acids and proteins using electrophoretic microfluidic devices, while pintsize devices have been developed for protein isolation according to miniaturized chromatography principles (size, charge, affinity, etc...
December 7, 2017: Analyst
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