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Gossett Daniel R - - 2010
Cell separation and sorting are essential steps in cell biology research and in many diagnostic and therapeutic methods. Recently, there has been interest in methods which avoid the use of biochemical labels; numerous intrinsic biomarkers have been explored to identify cells including size, electrical polarizability, and hydrodynamic properties. This review ...
Castillo Brandi - - 2010
TRPM7 is a ubiquitously expressed and constitutively active divalent cation channel essential for cell survival and proliferation because it provides a mechanism for Mg2+ entry. This makes the channel an attractive target for proliferative diseases. In keeping with its role in Mg2+ homeostasis, TRPM7 is inhibited by intracellular Mg2+ and ...
McQuade Lindsey E - - 2010
We report the visualization of NO production using fluorescence in tissue slices of the mouse main olfactory bulb. This discovery was possible through the use of a novel, cell-trappable probe for intracellular nitric oxide detection based on a symmetric scaffold with two NO-reactive sites. Ester moieties installed onto the fluorescent ...
Zeng Yong - - 2010
High-throughput genetic and phenotypic analysis at the single cell level is critical to advance our understanding of the molecular mechanisms underlying cellular function and dysfunction. Here we describe a high-performance single cell genetic analysis (SCGA) technique that combines high-throughput microfluidic emulsion generation with single cell multiplex polymerase chain reaction (PCR). ...
Xu Wei - - 2010
Arrays of transparent, releasable micrometer-scale structures termed "microcups" were created for the purpose of patterning and isolating viable cells from small cell samples. Cells were captured by the microcups without the need for barriers or walls on the intervening substrate. Furthermore, in contrast to prior methods for creating cell arrays ...
Jardillier Ludwig L Department of Biological Sciences, University of Warwick, Coventry, - - 2010
Global estimates indicate the oceans are responsible for approximately half of the carbon dioxide fixed on Earth. Organisms < or =5 microm in size dominate open ocean phytoplankton communities in terms of abundance and CO(2) fixation, with the cyanobacterial genera Prochlorococcus and Synechococcus numerically the most abundant and more extensively ...
Polzer Hans - - 2010
Assessment of cell fate is indispensable to evaluate cell-based therapies in regenerative medicine. Therefore, a widely used technique is fluorescence labeling. A major problem still is the standardized, noninvasive, and reliable quantification of fluorescence intensity of adherent cell populations on single-cell level, since total fluorescence intensity must be correlated to ...
Cho Sung Hwan - - 2010
We demonstrate a high performance microfabricated FACS system with highly integrated microfluidics, optics, acoustics, and electronics. Single cell manipulation at a high speed is made possible by the fast response time (approximately 0.1 ms) of the integrated PZT actuator and the nozzle structure at the sorting junction. A Teflon AF-coated ...
Nakata Eiji - - 2010
We have successfully developed a new cell-permeable SNARF derivative that is activated inside the cell by ester hydrolysis and is notable for having reduced background fluorescence.
Brasen Jens Christian JC Neutrophil Signalling Group, School of Medicine, Cardiff University, Heath Park, Cardiff, United - - 2010
Photolytic uncaging of biologically-active molecules within cells is a powerful technique. However, the delivery of uncaging light into the cytosol can vary between cell types, individual cells of the same type, and different loci within an individual cell because of optical differences in absorbance and light-scattering properties of the cytoplasm. ...
Oshige Hideyuki - - 2010
Temozolomide (TMZ) has been accepted as a standard antitumor drug for glioma worldwide. Regarding its mechanism of action, there are quite a few analyses. In the present study, we investigated the cell-killing effect and mechanism of action of TMZ with flow cytometry using glioblastoma cell lines. Each cell line was ...
Lovchik Robert D - - 2010
We present a method for depositing cells in the microchambers of a sealed microfluidic device and establishing flow across the chambers independently and serially. The device comprises a transparent poly(dimethylsiloxane) (PDMS) microfluidic network (MFN) having 2 cell chambers with a volume of 0.49 microL, 6 microchannels for servicing the chambers, ...
Bryce Steven M - - 2010
A flow cytometric method for analyzing suspension cell cultures for micronucleus content has been previously reported (Avlasevich et al. [2006]: Environ Mol Mutagen 47: 56-66). The experiments described herein were undertaken to evaluate the compatibility of this method (In Vitro MicroFlow) with attachment cells. Initially, CHO-K1 cells were studied in ...
Zhao Hong - - 2010
We have recently reported an innovative approach to use charged fluorochromes such as propidium iodide (PI) in the real-time, dynamic cell viability assays. This study was designed to provide a mechanistic rationale for the kinetic assays using cell permeability markers. Uptake of PI by live cells, effect on the cell ...
Hoffman Gregory R - - 2010
The mTORC1 pathway is a central regulator of cell growth, and defective mTORC1 regulation plays a causative role in a variety of human diseases, including cancer, tumor syndromes such as the tuberous sclerosis complex (TSC) and lymphangioleiomyomatosis (LAM), and metabolic diseases such as diabetes and obesity. Given the importance of ...
Martin Elizabeth I - - 2010
Corticotropin-releasing factor (CRF) orchestrates the mammalian endocrine, autonomic, and behavioral stress response and has been implicated in the pathophysiology of illnesses ranging from irritable bowel syndrome to mood and anxiety disorders. CRF is produced and released from a variety of cell types, making it difficult to distinguish the specific role ...
Kato Tatsuya - - 2010
A stably transformed insect cell expression system is superior to a baculovirus expression system, since the expression system is sustained and there is no cell lysis, but the isolation of cell lines producing recombinant proteins is time-consuming and laborious. In this study, we developed a technique for the rapid isolation ...
Gautrot Julien E - - 2010
The control of the cell microenvironment on model patterned substrates allows the systematic study of cell biology in well defined conditions, potentially using automated systems. The extreme protein resistance of poly(oligo(ethylene glycol methacrylate)) (POEGMA) brushes is exploited to achieve high fidelity patterning of single cells. These coatings can be patterned ...
Gabriele Sylvain - - 2010
This article describes a simple and low-tech microfluidic method for single-cell experimentation, which permits cell selection without stress, cell manipulation with fine control, and passive self-exclusion of all undesired super-micronic particles. The method requires only conventional soft lithography microfabrication techniques and is applicable to any microfluidic single-cell circuitry. The principle ...
Ishii Satoshi - - 2010
Various single-cell isolation techniques, including dilution, micromanipulation, flow cytometry, microfluidics, and compartmentalization, have been developed. These techniques can be used to cultivate previously uncultured microbes, to assess and monitor cell physiology and function, and to screen for novel microbiological products. Various other techniques, such as viable staining, in situ hybridization, ...
Sintes Eva - - 2010
Heterotrophic nanoflagellates (HNFs) are an essential component of all aquatic microbial food webs, and yet the exploration of the numerical and single-cell responses of these organisms in mixed assemblages still represents a major technical challenge. LysoTracker Green staining combined with flow cytometry was recently proposed for the enumeration of aquatic ...
He Peng - - 2010
In this paper, the amount of ascorbic acid (AA) in single rat peritoneal mast cell was determined by the method of capillary electrophoresis (CE) with electrochemical detection (ED) at a carbon fiber microdisk bundle electrode. The CE-ED system and the single-cell injection system were rearranged to make the operation more ...
Seitz Patricia M - - 2010
Superoxide affects many normal and pathogenic cellular processes, and the detection of superoxide produced by cells is therefore of interest for potential therapeutic applications. To develop a high-throughput cell-based assay for the detection of extracellular superoxide production that could be run in a 384-well or 1536-well format, 2 luminescent reagents, ...
Lassailly Francois - - 2010
Determining how normal and leukemic stem cells behave in vivo, in a dynamic and noninvasive way, remains a major challenge. Most optical tracking technologies rely on the use of fluorescent or bioluminescent reporter genes, which need to be stably expressed in the cells of interest. Because gene transfer in primary ...
Yuan Yufeng Y Division of Hepatobiliary Surgery, Department of Surgery, Zhongnan Hospital, Wuhan University, Wuhan, - - 2010
Recurrence and metastasis are the major factors associated with the poor prognosis of hepatocellular carcinoma (HCC). It was confirmed that multiple chemokines and their receptors are related to the progression and metastasis of HCC. The aim of this research was to conduct an investigation into whether macrophage inflammatory protein-1alpha/CCL3, and ...
Passante E - - 2010
OBJECTIVE: RBL-2H3 cells express Toll-like receptors, including TLR4. This study aims to assess various aspects of the TLR4 pathway. METHODS: RBL-2H3 cells were indirectly stained for cell surface TLR4, 25 CD14 and intracellular MyD88 proteins and analysed through flow cytometry for single-colour staining. RESULTS: While TLR4-receptors are expressed in RBL-2H3 ...
Takao Daisuke - - 2010
In sea urchin spermatozoa, the energy required for flagellar motility depends only on the diffusional supply from proximal mitochondria, and thus the diffusion rate inside flagella is one of the most crucial factors limiting the practical size and design of the motile machinery. To determine the diffusion rates of materials ...
Tölle Angelika - - 2010
Prerequisites for successful flow cytometry investigations are specific antibodies labeled with appropriate fluorochromes and negligible autofluorescence of the untreated cells at the wavelength of interest. The aim of this study was (a) to characterize frequently used urological carcinoma cell lines with regard to their autofluorescence properties, (b) to demonstrate the ...
Rath Amber L - - 2010
Osteocytes compose 90-95% of all bone cells and are the mechanosensors of bone. In this study, the strain experienced by individual osteocytes resulting from an applied fluid flow shear stress was quantified and correlated to two biological responses measured in real-time within the same individual osteocytes: (1) the upregulation of ...
Kenny Daniel J - - 2010
A new type of mass analyzer is described, which allows low-resolution axial ion ejection to be obtained from a traveling wave based, stacked ring collision cell. Linking this ejection temporally with the scanning of the second quadrupole of a tandem quadrupole mass spectrometer provides an improvement in sampling duty cycle, ...
Chen Hong - - 2010
We have developed a microfluidic method to construct an array on a circular disk for staining assays. In this method, convenient centrifugal liquid pumping has been achieved within the spiral microchannels by disk rotation or spinning. Moreover, the liquids flowing in spiral channels effectively interact with the along-channel intercepted cell ...
Ebert Berit - - 2010
Metabolic phenotyping at cellular resolution may be considered one of the challenges in current plant physiology. A method is described which enables the cell type-specific metabolic analysis of epidermal cell types in Arabidopsis thaliana pavement, basal, and trichome cells. To achieve the required high spatial resolution, single cell sampling using ...
Alvarez Marco - - 2011
When cultured cells are treated with fluorescent organelle probes or photosensitizer agents, a characteristic redistribution of fluorescence in cell structures occurs frequently after light irradiation. It is currently assumed that such changes, referred to as relocalizations of the fluorescent compounds, represent an important aspect of the photodynamic process, which is ...
Müller Susann - - 2010
The still poorly explored world of microbial functioning is about to be uncovered by a combined application of old and new technologies. Bacteria, especially, are still in the dark with respect to their phylogenetic affiliations as well as their metabolic capabilities and functions. However, with the advent of sophisticated flow ...
Falzone N - - 2010
Evaluation of the acrosome reaction can shed light on the fertilising competence of spermatozoa. To eliminate false-positive results when evaluating the acrosome status of human sperm cells, two viability probes propidium iodide (PI) and 7-amino-actinomycin D (7-AAD) were compared for their ability to stain nonviable cells post-fixation and permeabilisation. Both ...
Chen Pu - - 2010
Microfluidic cell sorter allows efficient separation of small number of cells, which is beneficial in handling cells, especially primary cells that cannot be expanded to large populations. Here, we demonstrate a microfluidic fluorescence-activated cell sorter (muFACS) with a novel sorting mechanism, in which automatic on-chip sorting is realized by turning ...
Nascimento Camila F - - 2010
Migration, invasion and protease activity are essential for tumor progression and metastasis. Metastatic cells rely on invadopodia to degrade and invade extracellular matrix (ECM). Invadopodia are membrane protrusions with enzymes required for ECM degradation. These protrusions contain cortactin and membrane type 1 matrix metalloproteinase (MT1-MMP) superimposed to areas of digested ...
Puccinelli John P - - 2010
Screening biological readouts in cell culture are increasing in frequency and throughput. In such assays, cell types may be rare and reagents or compounds may be expensive often resulting in a reduced number of conditions and/or replicates. "Tubeless" microfluidics offers a method to reduce this burden, as has been previously ...
Park Joong Yull - - 2010
Conventional cell trapping methods using microwells with small dimensions (10-20 mum) are useful for examining the instantaneous cell response to reagents; however, such wells have insufficient space for longer duration screening tests that require observation of cell attachment and division. Here we describe a flow method that enables single cell ...
Alley Jennifer - - 2010
The chemokine receptor CXCR5 is predominantly expressed on mature B cells and follicular T-helper cells. CXCR5 and its ligand CXCL13 participate in ectopic germinal center formation at the inflammatory sites of multiple immune diseases such as rheumatoid arthritis, multiple sclerosis, and Sjogren's syndrome. Therefore, disrupting CXCL13-induced chemotaxis may be a ...
Swearingen Kristian E - - 2010
A common method for quantifying cell death is measuring the concentration of lactate dehydrogenase (LDH) released by cells as their membranes become unstable. In cells expressing green fluorescent protein (GFP), degradation of the cell membrane also results in the release of GFP into the surrounding supernatant. In this study, we ...
Bajaj Avinash - - 2010
A family of conjugated fluorescent polymers was used to create an array for cell sensing. Fluorescent conjugated polymers with pendant charged residues provided multivalent interactions with cell membranes, allowing the detection of subtle differences between different cell types on the basis of cell surface features. Highly reproducible characteristic patterns were ...
Yang Yang - - 2010
We propose and demonstrate a simple, ultra sensitive radio frequency (RF) sensor to detect a single yeast cell and distinguish its viability in a microfluidic channel. On-chip interference is used to cancel background probing signals to improve sensor sensitivity. Individual viable and nonviable yeast cells (approximately 5.83 +/- 0.85 microm ...
Landrum Marie - - 2010
There is a continuing need for bioprobes that are target-specific and combine speed of delivery with maintenance of normal cell behaviour. Towards this end, we are developing small pro-fluorescent molecules that provide such specificity through chemical activation by biomolecules. We have generated a set of BODIPY (boron dipyrromethane) fluorophores, including ...
Karshafian Raffi - - 2010
This work investigates whether the application of sonoporation is limited by the size of a macromolecule being delivered and by the ability of cells to proliferate following uptake. KHT-C cells in suspension were exposed to variations in ultrasound pressure (0-570 kPa) and microbubble shell-type (lipid and protein) at fixed settings ...
Fong Dunne - - 2010
The phytochemical curcumin, from the Indian spice turmeric, has many biological properties, including anti-inflammatory and anti-carcinogenic activities. We have examined the effects of curcumin on the rat C6 glioma cell line. Treated and control cells were analyzed by Hoechst 33342 dye and flow cytometry. We observed a decrease in the ...
Randlev Britta - - 2010
GVAX immunotherapy for prostate cancer is comprised of two genetically modified prostate cancer cell lines, CG1940 and CG8711, engineered to secrete granulocyte macrophage-colony-stimulating factor. As part of the matrix of potency assays, CG1940 and CG8711 are tested for the expression level of cell surface HER-2/neu using a quantitative flow cytometer ...
Franke T - - 2010
We describe a novel microfluidic cell sorter which operates in continuous flow at high sorting rates. The device is based on a surface acoustic wave cell-sorting scheme and combines many advantages of fluorescence activated cell sorting (FACS) and fluorescence activated droplet sorting (FADS) in microfluidic channels. It is fully integrated ...
Vincent Meghan E - - 2010
Rare cells can be difficult to analyze because they either occur in low numbers or coexist with a more abundant cell type, yet their detection is crucial for diagnosing disease and maintaining human health. In this tutorial review, we introduce the concept of microfluidic stochastic confinement for use in detection ...
Mohammadpour H A - - 2010
AIMS: Mycoplasma agassizii can cause upper respiratory tract disease in the threatened desert tortoise of the Southwestern United States. Two technical challenges have impeded critical microbiological studies of this microorganism: (i) its small size limits the use of light microscopy for cell counting and (ii) its extremely slow growth in ...
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