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Hirano Masahiko - - 2010
We have developed a regulator-free device that enables long-term incubation of mammalian cells for epi-fluorescence imaging, based on a concept that the size of sample to be gassed and heated is reduced to observation scale. A poly(dimethylsiloxane) block stamped on a coverslip works as a long-lasting supplier of CO(2)-rich gas ...
Shrestha Bindesh - - 2010
Analysis of biochemicals in single cells is important for understanding cell metabolism, cell cycle, adaptation, disease states, etc. Even the same cell types exhibit heterogeneous biochemical makeup depending on their physiological conditions and interactions with the environment. Conventional methods of mass spectrometry (MS) used for the analysis of biomolecules in ...
Savina Ariel - - 2010
Phagosomes are complex organelles that form after ingestion by phagocytic cells of pathogens, dying cells, or cell debris. Highly dynamic interactions of phagosomes first with endosomes and then with lysosomes lead to the maturation of phagosomes into phagolysosomes. Contrary to other phagocytes, which degrade ingested particles to amino acids, dendritic ...
Sage Daniel - - 2010
Fluorescent and bioluminescent time-lapse microscopy approaches have been successfully used to investigate molecular mechanisms underlying the mammalian circadian oscillator at the single cell level. However, most of the available software and common methods based on intensity-threshold segmentation and frame-to-frame tracking are not applicable in these experiments. This is due to ...
Geisinger A - - 2010
Mammalian spermatogenesis is nowadays still poorly understood at the molecular level, mainly due to the heterogeneous nature of testes, which contain a high number of different cell types, and to the lack of spermatogenic cell culture systems for in vitro studies. As a consequence, the development and/or application of methodological ...
Est?vez M Carmen - - 2010
Aptamers are DNA or RNA oligonucleotide sequences that selectively bind to their target with high affinity and specificity. They are obtained using an iterative selection protocol called SELEX. Several small molecules and proteins have been used as targets. Recently, a variant of this methodology, known as cell-SELEX, has been developed ...
Terazono Hideyuki - - 2010
A reversible cell labelling method has been developed for non-destructive and non-invasive cell labelling and purification. Our method uses high affinity single strand DNA (ssDNA) aptamers against surface exposed target molecules on cells. The aptamers are subsequently removed from the cell surface using DNase nuclease treatment. We exemplified our method ...
Amano Maho - - 2010
It has known that the glycosylation plays an important role in the biological states, such as development, aging, and diseases. Although genomic and proteomic approaches have been intensively studied for diagnosis and disease treatment, glycomics have been laggard compared to them due to the hardness of the purification procedure from ...
Youk Hyun - - 2010
Yeast cells in an isogenic population do not all display the same phenotypes. To study such variation within a population of cells, we need to perform measurements on each individual cell instead of measurements that average out the behavior of a cell over the entire population. Here, we provide the ...
Akasaka Eri - - 2010
BACKGROUND: The recent availability of alpha-1,3-galatosyltransferase knockout pigs has eliminated anti-Gal antibodies to the galalpha1-3gal (alphagal epitope) as the major barrier to xenotransplantation. These alphagal epitope-negative animals can also be produced by somatic cell nuclear transfer of cells overexpressing endo-beta-galactosidase (EndoGalC), an enzyme capable of digesting the alphagal epitope. For ...
Zwirglmaier Katrin - - 2010
Fluorescence in situ hybridization with rRNA targeted oligonucleotide probes is nowadays one of the core techniques in microbial ecology, allowing the identification and quantification of microbial cells in environmental samples in situ. Next to the classic FISH protocol, which uses fluorescently monolabelled probes, the more sensitive CARD-FISH (also known as ...
Kentner David - - 2010
Following the introduction of fluorescent protein tags, the application of fluorescence microscopy in microbial cell biology has advanced the field dramatically. We now understand that bacterial cells are not simple bags of enzymes but have complex internal structures, and that specific intracellular organization plays an important role in a number ...
Lamm Monica H - - 2010
Cell trafficking of carbon nanotubes (CNTs) is an area of scientific inquiry that has great implications in medicine, biosensing, and environmental science and engineering. The essence of this inquiry resides in the interaction of carbon nanostructures and cell membranes, regulated by the laws of molecular cell biology and the physiochemical ...
Hayashi Tetsutaro - - 2010
To achieve an integrated understanding of the stem cell system of planarians at both the cellular and molecular levels, we developed a new method by combining "fluorescent activated cell sorting (FACS) index sorting" analysis and single-cell reverse transcription-polymerase chain reaction (RT-PCR) to detect the gene expression and cell cycle state ...
Park Joong Yull - - 2010
Slow interstitial flow can lead to large changes in cell morphology. Since conventional biological assays are adapted to two-dimensional culture protocols, there is a need to develop a microfluidic system that can generate physiological levels of interstitial flow. Here we developed a system that uses a passive osmotic pumping mechanism ...
Arnold Larry W - - 2010
Modern flow cytometric cell sorters are all capable of so-called "high-speed sorting." However, there is confusion about exactly how fast a "high-speed" cell sorter can sort cells. There are many considerations in achieving the fastest sorting speed, as well as the highest quality sort results--cell recovery, purity, and functionality. This ...
Lee Dennis - - 2010
The need for new pharmacological agents is unending. Yet the drug discovery process has changed substantially over the past decade and continues to evolve in response to new technologies. There is presently a high demand to reduce discovery time by improving specific lab disciplines and developing new technology platforms in ...
Nielsen Jeppe Lund - - 2010
Fluorescence in situ hybridization (FISH) can be combined with a number of staining techniques to reveal the relationships between the microorganisms and their function in complex microbial systems with a single-cell resolution. In this chapter, we have focused on staining methods for intracellular storage compounds (polyhydroxyalkanoates, polyphosphate) and a measure ...
Basu Sreemanti - - 2010
Experimental and clinical studies often require highly purified cell populations. FACS is a technique of choice to purify cell populations of known phenotype. Other bulk methods of purification include panning, complement depletion and magnetic bead separation. However, FACS has several advantages over other available methods. FACS is the preferred method ...
Chiavaroli Stefano - - 2010
A BARRIER IN SCALING LABORATORY PROCESSES INTO AUTOMATED MICROFLUIDIC DEVICES HAS BEEN THE TRANSFER OF LABORATORY BASED ASSAYS: Where engineering meets biological protocol. One basic requirement is to reliably and accurately know the distribution and number of biological cells being dispensed. In this study, a novel optical counting technique to ...
Pearson Jeffrey P - - 2010
The colonic mucus barrier is the first line of defence that the underlying mucosa has against the wide range of potentially damaging agents of microbial, endogenous, and dietary origin that occur within the colonic lumen. The functional component of mucus is the secreted, polymeric glycoprotein mucin. The mucus barrier can ...
Xiao Yan - - 2010
The rapid growth of the nanotechnology industry and the wide application of various nanomaterials have raised concerns over their impact on the environment and human health. Yet little is known about the mechanism of cellular uptake and cytotoxicity of nanoparticles. An array of nanomaterials has recently been introduced into cancer ...
Valero Ana - - 2010
There is great interest in highly sensitive separation methods capable of quickly isolating a particular cell type within a single manipulation step prior to their analysis. We present a cell sorting device based on the opposition of dielectrophoretic forces that discriminates between cell types according to their dielectric properties, such ...
Nash Katherine - - 2010
Fluorescence microscopy of cells loaded with fluorescent, Ca(2+)-sensitive dyes is used for measurement of spatial and temporal aspects of Ca(2+) signaling in live cells. Here we describe the method used in our laboratories for loading suspensions of human sperm with Ca(2+)-reporting dyes and measuring the fluorescence signal during physiological stimulation. ...
Huang Wei E - - 2010
Raman microspectroscopy is a noninvasive, label-free, and single-cell technology for biochemical analysis of individual mammalian cells, organelles, bacteria, viruses, and nanoparticles. Chemical information derived from a Raman spectrum provides comprehensive and intrinsic information (e.g., nucleic acids, protein, carbohydrates, and lipids) of single cells without the need of any external labeling. ...
Dharmasiri Udara - - 2010
Efficient selection and enumeration of low-abundance biological cells are highly important in a variety of applications. For example, the clinical utility of circulating tumor cells (CTCs) in peripheral blood is recognized as a viable biomarker for the management of various cancers, in which the clinically relevant number of CTCs per ...
Sasaki Akira - - 2010
Conventional biochemical assays are performed via an averaging procedure with the lysate of a large number of target cells; however, the averaged data lose information regarding the heterogeneity of individual cells. For quantitative assay of single cells, it is necessary to isolate single cells, extract cellular components and detect extremely ...
Wu Chunhui - - 2010
The application of quantum dots (QDs) in various biomedical areas requires detailed studies of their toxicity. We report a new strategy for probing the biocompatibility of these nanocrystals, namely, a dynamic investigation of cellular uptake images, cell growth curves, metabolic activity changes, and apoptosis aspects of cadmium telluride QDs capped ...
Emre Nil - - 2010
Due to the inherent sensitivity of human embryonic stem cells (hESCs) to manipulations, the recovery and survival of hESCs after fluorescence-activated cell sorting (FACS) can be low. Additionally, a well characterized and robust methodology for performing FACS on hESCs using multiple-cell surface markers has not been described. The p160-Rho-associated coiled ...
Wagner Michael - - 2010
A test system for cell viability based on colony formation has been established and applied to high resolution fluorescence microscopy and single molecule detection. Living cells were irradiated either by epi-illumination or by total internal reflection (TIR) of a laser beam, and light doses where at least 90% of irradiated ...
Radtke Andrea L - - 2010
The prevailing paradigm of Salmonella enteropathogenesis based on monolayers asserts that Salmonella pathogenicity island-1 Type Three Secretion System (SPI-1 T3SS) is required for bacterial invasion into intestinal epithelium. However, little is known about the role of SPI-1 in mediating gastrointestinal disease in humans. Recently, SPI-1 deficient nontyphoidal Salmonella strains were ...
Sun Deping - - 2010
Quantum dots (QDs) have shown great development potential in noninvasive imaging and monitoring of cancer cells in vivo because of their unique optical properties. However, the key issue of whether or not QDs-labeled cancer cells affect the proliferation, apoptosis and in vivo tumorigenicity ability has not been reported. The primary ...
Ariga Junko - - 2010
High-resolution time-lapse imaging of living zebrafish larvae can be utilized to visualize how biological processes unfold (for review see (1)). Compound transgenic fish which express different fluorescent reporters in neighboring cell types provide a means of following cellular interactions (2) and/or tissue-level responses to experimental manipulations over time. In this ...
Lu Zhe - - 2010
Integrating single-cell manipulation techniques in traditional and emerging biological culture systems is challenging. Microfabricated devices for single cell studies in particular often require cells to be spatially positioned at specific culture sites on the device surface. This paper presents a robotic micromanipulation system for pick-and-place positioning of single cells. By ...
Quah Benjamin J C - - 2010
Carboxyfluorescein succinimidyl ester (CFSE) is an effective and popular means to monitor lymphocyte division. CFSE covalently labels long-lived intracellular molecules with the fluorescent dye, carboxyfluorescein. Thus, when a CFSE-labeled cell divides, its progeny are endowed with half the number of carboxyfluorescein-tagged molecules and thus each cell division can be assessed ...
Schneider Marlon R - - 2010
E-cadherin is a major component of adherens junctions. Impaired expression of E-cadherin in the small intestine and colon has been linked to a disturbed intestinal homeostasis and barrier function. Down-regulation of E-cadherin is associated with the pathogenesis of infections with enteropathogenic bacteria and Crohn's disease. To genetically clarify the function ...
Langlois Marie-Josée - - 2010
The PTEN phosphatase acts on phosphatidylinositol 3,4,5-triphosphates resulting from phosphatidylinositol 3-kinase (PI3K) activation. PTEN expression has been shown to be decreased in colorectal cancer. Little is known however as to the specific cellular role of PTEN in human intestinal epithelial cells. The aim of this study was to investigate the ...
Dolowschiak Tamas - - 2010
The epithelium efficiently attracts immune cells upon infection despite the low number of pathogenic microbes and moderate levels of secreted chemokines per cell. Here we examined whether horizontal intercellular communication between cells may contribute to a coordinated response of the epithelium. Listeria monocytogenes infection, transfection, and microinjection of individual cells ...
Bassnett Steven - - 2010
The anterior face of the mouse lens is covered by a layer of epithelial cells. The epithelial cells serve a barrier function at the lens surface and as a progenitor population from which lens fiber cells, the predominant cell type of the lens, are derived. Decreased epithelial cell density is ...
Zhang Xiaoxiao X University of Hawaii at Manoa (UHM), Honolulu, HI 96822, USA. - - 2010
A microfluidic device was designed having the ability to continuously produce monodisperse microcapsules with controlled cell loading. The design included stages of inertial focusing, droplet generation, and photopolymerization. Prototype microfluidic devices were fabricated in polydimethylsiloxane (PDMS) to demonstrate each stage using poly(ethylene-glycol)-diacrylate (PEGDA) as the encapsulating material and oil as ...
Song Min Jae MJ Department of Biomedical Engineering, Case Western Reserve University, Cleveland, Ohio, - - 2010
A major hurdle to understanding and exploiting interactions between the stem cell and its environment is the lack of a tool for precise delivery of mechanical cues concomitant to observing sub-cellular adaptation of structure. These studies demonstrate the use of microscale particle image velocimetry (μ-PIV) for in situ spatiotemporal mapping ...
Bargmann Bastiaan O R BO Department of Biology, New York University, Center for Genomics and Systems Biology, - - 2010
High-resolution, cell type-specific analysis of gene expression greatly enhances understanding of developmental regulation and responses to environmental stimuli in any multicellular organism. In situ hybridization and reporter gene visualization can to a limited extent be used to this end but for high resolution quantitative RT-PCR or high-throughput transcriptome-wide analysis the ...
Zhang Jing - - 2010
Fluorophore-labeled bioprobes are the key for fluorescent-labeled imaging technology. In the present work, mouse liver hepatoma cell line BNL 1ME A.7R.1 (MEAR)-specific ssDNA aptamer TLS9a was used to fabricate quantum dot-labeled aptamer bioprobe (QD-Apt), which was obtained by conjugating streptavidin-modified quantum dots (SA-QDs) with biotin-derived aptamer via the interaction between ...
Saint-Ruf Claude - - 2010
We have developed a new method for accurate quantification of dead microbial cells. This technique employs the simultaneous use of fluorescent hydrazides and nucleic acid dyes. Fluorescent hydrazides allow detection of cells that cannot be detected with currently used high-affinity nucleic acid dyes. This is particularly important for nongrowing bacterial ...
Iijima Sou - - 2010
A new method for isolating targeted live bacterial cells was established with the use of cell sorting by flow cytometry (FCM) based on the fluorescence of the intermediate metabolite of biphenyl degradation. During biphenyl degradation, a PCB degrader, Comamonas testosteroni TK102, produces a meta-cleavage intermediate metabolite, 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid (HOPDA), which ...
Mogoa Emerancienne - - 2010
Chlorination is a well-known disinfection method, used in water treatment to inactivate various microorganisms, it induces numerous cellular changes. Even though Acanthamoebae are frequently found in water, the cellular changes induced in Acanthamoebae have not been described in the literature. Acanthamoebae are pathogenic amoebae and may provide a reservoir for ...
Wang Hu - - 2010
Cell penetrating peptides (CPPs) are promising tools for transducing presynthesized therapeutic molecules which possess low membrane permeability. The poor efficiency of cellular uptake and unexpected cellular localization are still the main obstacles to the development of drug delivery by using CPPs. In this study, we investigated the effect of a ...
Nyk?nen Marjukka - - 2010
We assessed morphology and ERalpha protein and gene expression of two breast cancer cell lines after three different fixatives: neutral-buffered 10% formaldehyde, LN-FIX and FineFIX and varying fixation times. We found that the cell morphology was best preserved in cells fixed with LN-FIX. Two commercial fixatives used in this study ...
Gassei Kathrin - - 2010
BACKGROUND: Isolation of spermatogonial stem cells (SSCs) could enable in vitro approaches for exploration of spermatogonial physiology and therapeutic approaches for fertility preservation. SSC isolation from adult testes is difficult due to low cell numbers and lacking cell surface markers. Glial cell-derived neurotrophic factor family receptor alpha-1 (GFRalpha1) plays a ...
de la Escosura-Muñiz Alfredo - - 2009
There is a high demand for simple, rapid, efficient, and user-friendly alternative methods for the detection of cells in general and, in particular, for the detection of cancer cells. A biosensor able to detect cells would be an all-in-one dream device for such applications. The successful integration of nanoparticles into ...
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