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Lulevich Valentin V Department of Chemistry, University of California, Davis, California 95616, - - 2009
Cell tracing dyes are very frequently utilized in cellular biology research because they provide highly sensitive fluorescent tags that do not compromise cellular functions such as growth and proliferation. In many investigations concerning cellular adhesion and mechanics, fluorescent dyes have been employed with the assumption of little impact on the ...
Fooksman David Robert - - 2009
Little is known about the signaling that occurs in an APC during contact with a T cell. In this article we report the concentration of the signaling lipid phosphatidylinositol 4,5-bisphosphate (PI(4,5)P(2)) at the APC side of the immunological synapse. In both human and mouse cells, a PI(4,5)P(2)-specific fluorescent reporter, PH-GFP ...
Buckmaster Ryan - - 2009
We report on a cell-based biosensor application that utilizes patterned single-cell arrays combined with confocal Raman spectroscopy to observe the time-dependent drug response of individual cells in real time. The patterned single-cell platform enables individual cells to be easily located and continuously addressable for Raman spectroscopy characterization of biochemical compositional ...
Granieri Lucia - - 2009
OBJECTIVES: To develop a high throughput screening-compatible assay for the selection of species-specific antibiotics that do not harm human cells. METHODS: Staphylococcus aureus and human reporter cells continuously generating a fluorescence signal were competitively co-cultivated. The fluorescence signals were determined in the presence and absence of the specific antibiotic streptomycin ...
Pina Ana - - 2009
One factor that contributes to a successful fruit tree grafting is the establishment of symplasmic contacts in the graft interface to facilitate the transfer of compounds between scion and stock. Using novel experimental and theoretical approaches we investigated whether the localized incompatibility, experienced in some Prunus grafts, could be related ...
Lähdesmäki Ilkka - - 2009
This work introduces, for the first time, simultaneous monitoring of fluorescence and absorbance using Bead Injection in a Lab-on-valve format. The aim of the paper is to show that when the target species, cells immobilized on a stationary phase, are exposed to reagents under well-controlled reaction conditions, dual monitoring yields ...
Baret Jean-Christophe - - 2009
We describe a highly efficient microfluidic fluorescence-activated droplet sorter (FADS) combining many of the advantages of microtitre-plate screening and traditional fluorescence-activated cell sorting (FACS). Single cells are compartmentalized in emulsion droplets, which can be sorted using dielectrophoresis in a fluorescence-activated manner (as in FACS) at rates up to 2000 droplets ...
Luo Chunxiong - - 2009
We present a simple and high-throughput microfluidic system for diffusion-based monolayer yeast cell culture monitoring. Yeast cells are patterned into the micro-cavity array with a suitable height (4 mum) that keeps the cells fixed in monolayer during the cell division. Different sizes of cavities and different repeating times of injection ...
Strovas Tim J - - 2009
Heterogeneity of cells within exponentially growing populations was addressed in a bacterium, the facultative methylotroph Methylobacterium extorquens AM1. A transcriptional fusion between a well-characterized methanol-inducible promoter (P(mxaF)) and gfp(uv) was used with flow cytometry to analyse the distribution of gene expression in populations grown on either succinate or methanol, correlated ...
Sankin Georgy - - 2009
Coupled oscillation of two laser generated microbubbles (maximum radius = 28 mum) and associated shear stresses are investigated experimentally. Bubble-bubble interaction in a microchannel of 25 mum height is observed using high-speed video cameras and muPIV technique. Two liquid microjets moving in opposite directions can be generated when the second ...
Tajiri Kazuto - - 2009
The analysis of single cells with multiple parameters in flow cytometry or microscopy requires suitable combinations of fluorophores and optical filters. The growing demands for the multiplex analysis of cells increase the requirements for developing new fluorophores and techniques. We have developed a novel method of analyzing a large number ...
Hsieh Ming-Fa - - 2009
We report the fluorescent labeling of osteoblast cells using the biocompatible hydroxyapatite (HA) grown with nucleating seed of hydrophilic CdSe/ZnS quantum dots (QDs) allowing the real-time observation of cell under confocal microscope. We found that the MC3T3-E1 osteoblast cells can engulf HA with surface-tailored QDs showing fluorescent spots in the ...
Hennig Christian - - 2009
Analysis of the immense complexity of the immune system is increasingly hampered by technical limitations of current methodologies, especially for multiparameter- and functional analysis of samples containing small numbers of cells. We here present a method, which is based on the stepwise functional manipulation and analysis of living immune cells ...
Crow Matthew J - - 2009
OBJECTIVE: The goal of this study was to assess whether immunolabeled nanoparticle biomarkers are comparable to fluorescent marker imaging in measuring epidermal growth factor receptor (EGFR) expression. MATERIALS AND METHODS: EGFR expression was quantified using both imaging methods in four cell lines: A431 human epidermoid carcinoma cells, which are known ...
Mukai Rie - - 2009
Flavonoids are widely distributed in the plant kingdom and show various biological activities. The bioavailability of flavonoids in biological samples has conventionally been quantified by high-performance liquid chromatography and mass spectrometry, but with these analytical techniques it is difficult to estimate the subcellular localization of flavonoids in intact cells. In ...
Schütz Christian - - 2009
Novel immunosuppressive strategies are targeting for an antigen-specific deletion of T cells responsible for organ damage in autoimmunity and allograft rejection. Here, we present a new flow cytometry-based assay that allows the reliable and efficient detection of T cells that were eliminated in an antigen-specific fashion. A stable cell-labelling technique ...
Vandewoestyne Mado - - 2009
Laser microdissection is a valuable tool for isolating specific cells from mixtures, such as male cells in a mixture with female cells, e.g., in cases of sexual assault. These cells can be stained with Y-chromosome-specific probes. We developed an automatic screening method to detect male cells after fluorescence in situ ...
Jiang Shan - - 2009
Near-infrared (NIR)-to-visible upconversion fluorescent nanoparticles were synthesized and used for imaging and targeted delivery of small interfering RNA (siRNA) to cancer cells. Silica-coated NaYF(4) upconversion nanoparticles (UCNs) co-doped with lanthanide ions (Yb/Er) were synthesized. Folic acid and anti-Her2 antibody conjugated UCNs were used to fluorescently label the folate receptors of ...
Schmidt Enrico K - - 2009
We developed a nonradioactive fluorescence-activated cell sorting-based assay, called surface sensing of translation (SUnSET), which allows the monitoring and quantification of global protein synthesis in individual mammalian cells and in heterogeneous cell populations. We demonstrate here, using mouse dendritic and T cells as a model, that SUnSET offers a technical ...
Wlodkowic Donald D Department of Electronics and Electrical Engineering, The Bioelectronics Research Center, University of Glasgow, Glasgow, UK. - - 2009
Cell death is a stochastic process, often initiated and/or executed in a multi-pathway/multi-organelle fashion. Therefore, high-throughput single-cell analysis platforms are required to provide detailed characterization of kinetics and mechanisms of cell death in heterogeneous cell populations. However, there is still a largely unmet need for inert fluorescent probes, suitable for ...
Song Er-Qun - - 2009
BACKGROUND: Luminescent nanobioprobes with cell-targeting specificity are likely to find important applications in bioanalysis, biomedicine, and clinical diagnosis. Quantum dots (QDs) are unique and promising materials for such a purpose because of their fluorescence and large surface area for attaching cell-targeting molecules. METHODS: We produced water-dispersible QDs by coating hydrophobic ...
Pijanka Jacek Klaudiusz - - 2009
Single-cell studies have important implications in biomedicine. An accurate investigation of biochemical behaviour and status requires a biomolecular probe such as vibrational microscopy. Amongst other approaches, synchrotron infrared microspectroscopy is an appropriate analytical tool for single-cell investigation. However, it is important to understand the precise origin of spectral differences as ...
Malone Cheryl L - - 2009
With the emergence of Staphylococcus aureus as a prominent pathogen in community and healthcare settings, there is a growing need for effective reporter tools to facilitate physiology and pathogenesis studies. Fluorescent proteins are ideal as reporters for their convenience in monitoring gene expression, performing host interaction studies, and monitoring biofilm ...
Dobson Joseph T - - 2009
The zebrafish is a robust animal model for studying vertebrate haematopoiesis and immune cell interactions. However, fluorescence activated cell sorting (FACS) has been limited due to a paucity of available functional zebrafish antibodies. We have developed a technique combining FACS with whole mount in situ hybridization (WISH) that enables the ...
Roth Thomas - - 2009
I(SC)-V(OC) curves measured by the suns-V(OC) method are widely used for solar cell characterization due to its being unaffected by series resistance effects. A common setup for this measurement system uses a xenon photoflash for illumination purposes, resulting in a fast acquisition of the suns-V(OC) measurement data during the decaying ...
Olofsson Jessica - - 2009
Methods that can control and vary the solution environment around single cells are abundant. In contrast, methods that offer direct access to the intracellular proteome and genome in single cells with the control, flexibility, and convenience given by microfluidic methods are both scarce and in great demand. Here, we present ...
Maki Chad B - - 2009
BACKGROUND: Knowledge about the identity and characteristics of spermatogonial stem cells (SSCs) in human is very limited. Here, Rhesus monkey was used as an animal model to investigate molecular and phenotypic characteristics of SSCs in the adult testes. METHODS: A variety of immunohistological, molecular biological and functional assays were used ...
Wilke, Lars Martin
An algorithm is presented which fills contours described by a set of pixels whose locations are given in unsorted linear quadtree (LQT) form. Each pixel requires an associated blocking code which indicates locally in which direction the region should grow. The technique takes advantage of certain characteristics of LQT location ...
Lovchik Robert D - - 2009
We present a method for depositing cells in a sealed microfluidic device. The device consists of a poly(dimethylsiloxane) (PDMS) microfluidic network (MFN) sealed with a Si chip. The Si chip has vias and ports that are connected to high-precision motorized pumps. The surfaces of the PDMS MFN are homogeneously coated ...
Murata Masaya - - 2009
We investigated properties of cells affecting their optical trapping force and successfully established a novel cell separation method based on the combined use of optical trapping force and microfluidics on a microchip. Our investigations reveal that the morphology, size, light absorption, and refractive index of cells are important factors affecting ...
Da Silveira M G - - 2009
AIMS: To study the effect of ethanol on Oenococcus oeni activity at the single cell level. METHODS AND RESULTS: The active extrusion of the fluorescent probe carboxy fluorescein (cF) was used to assess the metabolic activity of ethanol-stressed O. oeni cells. Subsequent flow cytometric analysis revealed that O. oeni cells ...
Fritzsche Michael - - 2009
A novel cell-based fluorometric sensor system for toxicity monitoring is described, which uses functional spontaneously contracting cardiomyocytes (HL-1 cell line) as the biological recognition element. Based on these highly specialized cells, it has the potential of providing a sensitive and relevant analytical in vitro toxicity testing method. The system was ...
Mahringer Anne - - 2009
Purpose: To establish a fluorescence-based assay for drug interactions with the ABC-export-protein BCRP (ABCG2). Methods: BCRP expression was verified by immunostaining and Western blots in intact porcine brain capillaries, isolated endothelial cells (PBCECs) and MDCKII-cells over-expressing human wild-type BCRP (MDCKII-hBCRP). Transport of fluorescent mitoxantrone across cells was determined to assess ...
Duhamel Solange - - 2009
ELF97 phosphate (ELF-P) is a useful compound for assessing the phosphorus-related status of planktonic aquatic populations. The technique has been successfully applied to phytoplankton and more recently to heterotrophic prokaryotes in both freshwater and marine samples. We have used a recently developed protocol that enables the detection by flow cytometry ...
Misiewicz-KrzemiƄska I - - 2009
The ability of protoporphyrin IX (PP IX) to enter neoplastic cells is used for diagnosis and photodynamic therapy. We present here the effects of PP IX and three amino acid derivatives on MCF-7 cell viability. The fluorescence of each substance was examined using confocal microscopy. Our results show that diarginine ...
Xie Wenjun - - 2009
A capillary electrophoresis (CE) method based on an enzymatic cycling reaction is developed to determine both NAD(+) and NADH in a single cell in a single run. The detection limit can reach down to 0.2 amol of NAD(+) and 1 amol of NADH with a homemade capillary electrophoresis laser-induced fluorescence ...
Harz M - - 2009
Rapid microbial detection and identification with a high grade of sensitivity and selectivity is a great and challenging issue in many fields, primarily in clinical diagnosis, pharmaceutical, or food processing technology. The tedious and time-consuming processes of current microbiological approaches call for faster ideally on-line identification techniques. The vibrational spectroscopic ...
Kortmann Hendrik - - 2009
Single cell analysis is mainly limited to single time-point measurements, without the possibility to track behavioral changes of a single cell or its descendents. Here, the integration of a spatiotemporal single cell lab-on-a-chip system with an automated cultivation device allows single cell analysis under defined growth conditions and, especially, semiautomated ...
Phillips Joseph A - - 2009
This work describes the development and investigation of an aptamer modified microfluidic device that captures rare cells to achieve a rapid assay without pretreatment of cells. To accomplish this, aptamers are first immobilized on the surface of a poly(dimethylsiloxane) microchannel, followed by pumping a mixture of cells through the device. ...
Barhoumi Rola - - 2009
Quantification of polycyclic aromatic hydrocarbons (PAH) and their metabolites within living cells and tissues in real time using fluorescence methods is complicated due to overlaping excitation and/or emission spectra of metabolites. In this study, simultaneous analysis of several metabolites of a prototype carcinogenic PAH, benzo[a]pyrene (BaP) in undifferentiated (MCF10A) and ...
Fong Chui Yee - - 2009
A major concern in human embryonic stem cell (hESC)-derived cell replacement therapy is the risk of tumorigenesis from undifferentiated hESCs residing in the population of hESC-derived cells. Separation of these undifferentiated hESCs from the differentiated derivatives using cell sorting methods may be a plausible approach in overcoming this problem. We ...
Yan Jun - - 2009
The integration of sensing elements with small groups of cells is a critical step towards miniaturization of cell cultivation and analysis. This paper describes the development of an optical, enzyme-based biosensor for local detection of hydrogen peroxide (H(2)O(2)) secreted by stimulated macrophages. Photolithographic patterning of horseradish peroxidase (HRP)-containing poly (ethylene ...
Nyland Jennifer F - - 2009
Engineered nanoparticles (NPs) possess a range of biological activity. In vitro methods for assessing toxicity and efficacy would be enhanced by simultaneous quantitative information on the behavior of NPs in culture systems and signals of cell response. We have developed a method for visualizing NPs within cells using standard flow-cytometric ...
Vandewoestyne, Mado
Laser microdissection is a valuable tool for isolating specific cells from mixtures, such as male cells in a mixture with female cells, e.g., in cases of sexual assault. These cells can be stained with Y-chromosome specific probes. We developed an automatic screening method to detect male cells after fluorescence in ...
Hart Traver - - 2009
Here we describe human spotted cell chips, a technology for determining cellular state across arrays of cells subjected to chemical or genetic perturbation. Cells are grown and treated under standard tissue culture conditions before being fixed and printed onto replicate glass slides, effectively decoupling the experimental conditions from the assay ...
An W Frank - - 2009
Fluorescence-based assays are widely used in high-throughput screening due to their high sensitivity, diverse selection of fluorophores, ease of operation, and various readout modes. As a result, fluorescence-based assays have been applied to monitor a broad range of activities in life-science research such as molecular dynamics and interactions, enzymatic activities, ...
Dustin Michael L - - 2009
Recently implemented fluorescence imaging techniques, such as total internal reflection fluorescence microscopy and two-photon laser scanning microscopy, have made possible multiscale analysis of the immune response from single molecules in an interface to cells moving in lymphoid tissues and tumors. In this review, we consider components of T cell sensitivity: ...
Padfield Dirk - - 2009
A growing number of screening applications require the automated monitoring of cell populations in a high-throughput, high-content environment. These applications depend on accurate cell tracking of individual cells that display various behaviors including mitosis, occlusion, rapid movement, and entering and leaving the field of view. We present a tracking approach ...
Stephen Michael J - - 2009
Neonatal and adult cardiomyocytes were isolated from rat hearts. Some of the adult myocytes were cultured to allow for cell dedifferentiation, a phenomenon thought to mimic cell changes that occur in stressed myocardium, with myocytes regressing to a fetal pattern of metabolism and stellate neonatal shape.Using fluorescence deconvolution microscopy, cells ...
Nisato Riccardo E - - 2009
This chapter describes detailed methods for the isolation of primary human lymphatic endothelial cells from neonatal foreskin. We also provide protocols and information for their characterization and propagation. Isolation of primary human lymphatic endothelial cells requires a two-step process: mechanical and enzymatic digestion of human foreskins and cell sorting by ...
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