Relevant bibliographies by topics / Yeast fluorescence

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  1. Journal articles
  2. Dissertations / Theses
  3. Books
  4. Book chapters
  5. Conference papers

Academic literature on the topic 'Yeast fluorescence'

Author: Grafiati
Published: 4 June 2021
Last updated: 1 February 2022

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Journal articles on the topic "Yeast fluorescence"

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Karreman, Robert J., and George G. Lindsey. "A Rapid Method to Determine the Stress Status of Saccharomyces cerevisiae by Monitoring the Expression of a Hsp12:Green Fluorescent Protein (GFP) Construct under the Control of the Hsp12 Promoter." Journal of Biomolecular Screening 10, no. 3 (April 2005): 253–59. http://dx.doi.org/10.1177/1087057104273485.

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The gene for the green fluorescent protein (GFP) was fused in-frame to the 3′ end of HSP12. This construct was regulated by the HSP12 promoter in a pYES2 yeast expression vector. No fluorescence was observed in yeast growing exponentially in glucose-containing medium, but fluorescence was observed when the yeast entered the stationary phase. Fluorescence microscopy indicated that the fusion protein was localized to the peripheral regions of the cell as well as to the cytoplasm and the tonoplast. Subjecting the yeast to a variety of stresses known to induce HSP12 transcription, including salt, osmotic, ethanol, and heat stress, resulted in a time-dependent increase in GFP fluorescence. The use of this system as a method to assess the general stress status of yeast growing in an industrial application is proposed.
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Jones, Laura M., Danielle Dunham, Monique Y. Rennie, Jeffrey Kirman, Andrea J. Lopez, Klara C. Keim, William Little, et al. "In vitro detection of porphyrin-producing wound bacteria with real-time fluorescence imaging." Future Microbiology 15, no. 5 (March 2020): 319–32. http://dx.doi.org/10.2217/fmb-2019-0279.

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Aim: Fluorescence imaging can visualize polymicrobial populations in chronic and acute wounds based on porphyrin fluorescence. We investigated the fluorescent properties of specific wound pathogens and the fluorescence detected from bacteria in biofilm. Methods: Utilizing Remel Porphyrin Test Agar, 32 bacterial and four yeast species were examined for red fluorescence under 405 nm violet light illumination. Polymicrobial biofilms, supplemented with δ-aminolevulinic acid, were investigated similarly. Results: A total of 28/32 bacteria, 1/4 yeast species and polymicrobial biofilms produced red fluorescence, in agreement with their known porphyrin production abilities. Conclusion: These results identify common wound pathogens capable of producing porphyrin-specific fluorescence and support clinical observations using fluorescence imaging to detect pathogenic bacteria in chronic wounds.
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Rosseau, S., W. Seeger, H. Pralle, and J. Lohmeyer. "Phagocytosis of viable Candida albicans by alveolar macrophages: flow cytometric quantification." American Journal of Physiology-Lung Cellular and Molecular Physiology 267, no. 2 (August 1, 1994): L211—L217. http://dx.doi.org/10.1152/ajplung.1994.267.2.l211.

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The phagocytic capacity of blood leukocytes may be assessed by flow cytometric techniques using fluorochrome-labeled particles including viable microorganisms. Application of this approach to alveolar macrophages (AM) is hampered or even rendered impossible by the strong autofluorescence of this cell type, superimposing the fluorescence intensity of the labeled phagocytic targets. Viable Candida albicans were loaded with the membrane-permeable fluorescent dye carboxy-seminaphtorhodafluor 2/acetoxymethylester (carboxy-SNARF 2-AM), which is cleaved intracellularly to generate the membrane-impermeable derivative carboxy-SNARF 2. Fluorescence was excited with the 488-nm line of an argon-ion laser, and the emission peak at 633 nm was used for quantification of dye-associated fluorescence. Rabbit and human AM were labeled with fluorescein isothiocyanate-coupled monoclonal mouse anti-macrophage antibodies. After coincubation of macrophages and yeast, 4% paraformaldehyde plus 0.5% EDTA in phosphate-buffered saline was used to stop the phagocytic process and detach adherent yeast from the AM surface. Macrophages loaded with yeast displayed a shift from monochromatic (green) to dual (green and red) fluorescence. The percentage of yeast-positive AM and red fluorescence intensity of phagocytosing macrophages were quantified. Yeast opsonization with serum or anti-Candida immunoglobulins was a prerequisite for phagocytosis. Under optimized conditions (0.5-10% serum; 60 min yeast-AM incubation; yeast-AM ratio 8:1 to 12:1), 71-91% of the AM were involved in the phagocytic process. Yeast engulfment was completely inhibited by N-ethylmaleimide and iodoacetic acid.(ABSTRACT TRUNCATED AT 250 WORDS)
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Thiebault, F., and J. Coulon. "Influence of carbon source and surface hydrophobicity on the aggregation of the yeastKluyveromyces bulgaricus." Canadian Journal of Microbiology 51, no. 1 (January 1, 2005): 91–94. http://dx.doi.org/10.1139/w04-106.

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Aggregation of the yeast Kluyveromyces bulgaricus is mediated by the galactose-specific lectin KbCWL1. This lectin contains hydrophobic amino acids and its activity is calcium dependent. A specific fluorescent probe, 1-anilinonaphthalene-8-sulfonic acid in the free acid form (ANS; Sigma Chemical Co., St. Louis, Missouri), was used to study the hydrophobic areas on the cellular surface of K. bulgaricus. Changes in surface hydrophobicity during the growth and aggregation of yeast cells were studied. Surface hydrophobicity increased during growth and depended on the amount of yeast cells in the culture medium. During growth, the size of the hydrophobic areas on the cell surface was measured using ANS and was found to increase with the percentage of flocculating yeasts. Our results strongly suggest that the hydrophobic areas of the cell walls of yeast cells are involved in the aggregation of K. bulgaricus.Key words: aggregation, carbon source, fluorescence probe, hydrophobicity, yeast.
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Skruzny, Pohl, and Abella. "FRET Microscopy in Yeast." Biosensors 9, no. 4 (October 11, 2019): 122. http://dx.doi.org/10.3390/bios9040122.

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Förster resonance energy transfer (FRET) microscopy is a powerful fluorescence microscopy method to study the nanoscale organization of multiprotein assemblies in vivo. Moreover, many biochemical and biophysical processes can be followed by employing sophisticated FRET biosensors directly in living cells. Here, we summarize existing FRET experiments and biosensors applied in yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe, two important models of fundamental biomedical research and efficient platforms for analyses of bioactive molecules. We aim to provide a practical guide on suitable FRET techniques, fluorescent proteins, and experimental setups available for successful FRET experiments in yeasts.
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Audus, K. L., M. R. Tavakoli-Saberi, H. Zheng, and E. N. Boyce. "Chlorhexidine Effects on Membrane Lipid Domains of Human Buccal Epithelial Cells." Journal of Dental Research 71, no. 6 (June 1992): 1298–303. http://dx.doi.org/10.1177/00220345920710060601.

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The effect of chlorhexidine gluconate on the adherence of Candida albicans to human buccal epithelial cells (BEC) and drug-induced alterations in BEC membrane-lipid packing order were examined. Treatment of BEC with attached yeasts with 0.1 and 0.2% chlorhexidine resulted in significant yeast detachment after 90 and 60 min, respectively. Following pre-treatment of BEC with > 0.1% chlorhexidine, yeast adherence was inhibited by > 80%. In parallel experiments, the fluorescence anisotropy of BEC labeled with fluorescent membrane probes-diphenylhexatriene (DPH) and trimethylammonium DPH-was assessed following exposure to chlorhexidine. The fluorescence anisotropy decreased with increasing concentrations of chlorhexidine, which indicated that the drug decreased epithelial-cell membrane-lipid packing order. Chlorhexidine concentrations that altered epithelial-cell membrane-lipid packing order, particularly in superficial regions, were similar to those drug concentrations required for detachment of adherent yeasts. Similar results were obtained with a second antifungal, nystatin A. While the effects of chlorhexidine on the buccal-cell membrane-lipid packing order were not reversed by multiple washings, the opposite situation occurred with nystatin A. The results suggest that chlorhexidine-induced alterations ofBEC membrane-lipid order may be involved in the antifungal actions of the drug.
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Guenther, Margarita, Falko Altenkirch, Kai Ostermann, Gerhard Rödel, Ingo Tobehn-Steinhäuser, Steffen Herbst, Stefan Görlandt, and Gerald Gerlach. "Optical and impedimetric study of genetically modified cells for diclofenac sensing." Journal of Sensors and Sensor Systems 8, no. 1 (May 21, 2019): 215–22. http://dx.doi.org/10.5194/jsss-8-215-2019.

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Abstract. Whole-cell biosensors, based on genetically modified yeast cells, were employed to detect anthropogenic micropollutants (e.g. drugs). Specific stimuli, e.g. traces of drugs, lead to the induction of fluorescence in the respective cells. Receptors of the cells detect specific signal molecules and induce the formation of fluorescent proteins. In this work, genetically modified cells of the yeast Saccharomyces cerevisiae BY4741 were confined in a four-chamber microfluidic cell, providing an optical monitoring of the cell behaviour and their supply with the nutrients. The measurements of the time-dependent fluorescence intensity were performed with different concentrations of the drug diclofenac, and the sensitivity of yeast cells to diclofenac was demonstrated. Cell viability was monitored by simultaneous impedance recording.
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Vanek, Martin, Filip Mravec, Martin Szotkowski, Dana Byrtusova, Andrea Haronikova, Milan Certik, Volha Shapaval, and Ivana Marova. "Fluorescence lifetime imaging of red yeast Cystofilobasidium capitatum during growth." EuroBiotech Journal 2, no. 2 (April 1, 2018): 114–20. http://dx.doi.org/10.2478/ebtj-2018-0015.

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AbstractRed yeast Cystofilobasidium capitatum autofluorescence was studied by means of confocal laser scanning microscopy (CLSM) to reveal distribution of carotenoids inside the cells. Yeasts were cultivated in 2L fermentor on glucose medium at permanent light exposure and aeration. Samples were collected at different times for CLSM, gravimetric determination of biomass and HPLC determination of pigments. To compare FLIM (Fluorescence Lifetime Imaging Microscopy) images and coupled data (obtained by CLSM) with model systems, FLIM analysis was performed on micelles of SDS:ergosterol and SDS:coenzyme Q with different content of ergosterol and coenzyme Q, respectively, and with constant addition of beta-carotene. Liposomes lecithin:ergosterol:beta-carotene were investigated too. Two different intracellular forms of carotenoids were observed during most of cultivations, with third form appeared at the beginning of stationary phase. Observed behavior is probably due to formation of some kind of carotenoid protective system in membranes of different compartments of yeast cell, especially cytoplasmic membrane.
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Stasyuk, Natalia Ye, Galina Z. Gayda, Roman Ja Serkiz, and Mykhailo V. Gonchar. "Cell Imaging with Fluorescent Bi-Metallic Nanoparticles." JOURNAL OF ADVANCES IN CHEMISTRY 11, no. 4 (March 9, 2015): 3499–511. http://dx.doi.org/10.24297/jac.v11i4.6694.

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Last decades various imaging techniques have been applied in biological and biomedical research, such as magnetic resonance imaging, different types of tomography, fluorescence/bioluminescence, ultrasound, as well as multimodality approaches. Fluorescence imaging, especially in combination with nanoscale materials, is a very prospective tool for experiments in vivo and clinical applications due to its high temporal and spatial resolutions. Fluorescent nanoparticles (NPs), having ability to interact with biomolecules both on the surface of and inside the cells, may revolutionize the cell imaging approaches for diagnostics and therapy. In our investigation we report about new method of cell imaging with fluorescent bi-metallic NPs synthesized by chemical reduction of the relevant ions. As the model of living organism, the cells of yeast Hansenula polymorpha were used. All NPs in minimal concentration (up to 0.05 mM) was proved to be non-toxic for yeast cells. The NPs and NPs-modified cells were characterized with the methods of UV-VIS spectroscopy, scanning electron microscopy, atom force microscopy, transmission electron microscopy and fluorescence microscopy. The bimetallic NPs, possessing the stable fluorescence in solution and inside the cells, allow to observe the phenomenon of NPs transferring from parental to daughter cells through at least three generations followed by releasing from the modified cells. The fluorescent NPs synthesized being small, non-toxic and fluorescent was shown to be perspective tool for cell imaging.
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Monosov, E. Z., T. J. Wenzel, G. H. Lüers, J. A. Heyman, and S. Subramani. "Labeling of peroxisomes with green fluorescent protein in living P. pastoris cells." Journal of Histochemistry & Cytochemistry 44, no. 6 (June 1996): 581–89. http://dx.doi.org/10.1177/44.6.8666743.

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We exploited the light-activated fluorescent properties of the green fluorescent protein (GFP) of the jellyfish Aequorea victoria for studies on the peroxisomal sorting of polypeptides. GFP and GFP-SKL (containing a C-terminal, tripeptide peroxisomal targeting signal, SKL) were expressed from a methanol-inducible, alcohol oxidase (AOX1) promoter in the methylotrophic yeast Pichia pastoris. GFP was cytosolic, whereas the GFP-SKL fusion protein was targeted to peroxisomes, as demonstrated by biochemical fractionation of organelles on Nycodenz gradients. Neither GFP nor GFP-SKL affected the viability of yeast cells but both were fluorescent on excitation with 395-nm UV light. The subcellular locations of GFP and GFP-SKL in living yeast cells were monitored by fluorescence microscopy and their fluorescence was coupled to photo-oxidation of diaminobenzidine (DAB), resulting in the deposition of electron-dense oxidized DAB at intracellular locations of GFP derivatives. This photooxidation procedure permitted facile ultrastructural localization of GFP in cells by electron microscopy, and provided further evidence that GFP produced in P. pastoris is cytosolic, whereas GFP-SKL is peroxisomal. The GFP-SKL fusion protein is therefore a versatile reporter for the peroxisomal compartment, with many applications for studies involving peroxisomal import and biogenesis.
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Dissertations / Theses on the topic "Yeast fluorescence"

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Bakker, Elco. "Quantitative fluorescence microscopy methods for studying transcription with application to the yeast GAL1 promoter." Thesis, University of Edinburgh, 2016. http://hdl.handle.net/1842/20403.

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The advent and establishment of systems biology has cemented the idea that real understanding of biological systems requires quantitative models, that can be integrated to provide a complete description of the cell and its complexities. At the same time, synthetic biology attempts to leverage such quantitative models to efficiently engineer novel, predictable behaviour in biological systems. Together, these advances indicate that the future understanding and application of biology will require the ability to create, parameterise and discriminate between quantitative models of cellular processes in a rigorous and statistically sound manner. In this thesis we take the regulation of GAL1 expression in Saccharomyces cerevisiae as a test case and look at all aspects of this process: from reporter selection to data acquisition and statistical analysis. In chapter B we will discuss optimal fluorescent reporter selection and construction for investigating transcriptional dynamics, as well as procedures for quantifying and correcting the various sources of error in our microscope system. In chapter 3 we will describe software developed to analyse fluorescent microscopy images and convert them to gene expression data. A number of iterations of the software are tested against manually curated data sets, and the measurement error produced by its imperfections is quantified and discussed. In chapter 4 a method, based on fluctuations in photobleaching, is developed for quantifying both measurement error and the relationship between protein concentration and measured fluorescence. The method is refined and its efficacy discussed. In the last section I make a preliminary application of these methods to investigating the regulatory effect of the GAL10-lncRNA. Interesting phenomena are observed and further investigated using two new strains: genetic variants expressing a fluorescent reporter from the GAL1 promoter, one harbouring a wild type GAL1 promoter and one in which the binding site for the Gal10 noncoding RNA has been removed. The methods developed throughout the thesis are applied and the data analysed. A heterogeneous response, distinguishable between the two strains, is observed and related to cell-to-cell variations in growth rate.
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Jeřábková, Petra. "Studium vlastností biologického materiálu pomocí metod obrazové analýzy." Doctoral thesis, Vysoké učení technické v Brně. Fakulta chemická, 2010. http://www.nusl.cz/ntk/nusl-233311.

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Within the dissertation thesis “Study of Biological Material Attributes by Using Image Analysis Methods”, attention is focused on monitoring of the application of image analysis methods, mostly a fractal analysis, in studying the properties of various yeast species. The thesis includes determining the number of yeast cells and vegetative propagation of yeast using fractal parameters – fractal measure D and fractal dimension K. Attention is also paid not only to the application of the existing image analysis methods, but also to their renovation. The obtained images were evaluated using the box counting method specified by implementation of wavelet transformation. To monitor yeast cells for a longer time, it was first necessary to prepare a suitable microscopic preparation. To distinguish live and dead cells, the following fluorescent dyes were used: acridine orange, fluorescein diacetate, FUN-1, and Calcofluor White M2R. The images of yeast cells were recorded using a still camera or a CCD camera and microscope. Clips of the same size were obtained from the acquired digital photographs and processed by the HarFA program developed at the Faculty of Chemistry, Brno University of Technology. On the results it is possible to see a change in the fractal dimension depending on time, i.e. on the change of a budding cell structure, or to determine the number and radius of yeast cells upon predefined conditions.
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Lichten, Catherine Anne. "Quantitative fluorescence methods for studying cellular protein networks, with applications to the yeast galactose pathway." Thesis, McGill University, 2013. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=114175.

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Biology in the past two decades has shifted away from qualitative observation towards quantitative data, and away from reductionism towards viewing the phenomena under study as systems. Developments in fluorescent reporters, experimental instrumentation, and computational power have been integral to this change. A key feature of this transformation has been a substantial increase in the use of mathematical models to explore and verify our understanding of complicated biological mechanisms. This thesis looks at three different aspects of the systems modelling approach: data acquisition, model creation and evaluation, and measurement of parameter values. The system under study for the first two parts is the GAL pathway of the yeast Saccharomyces cerevisiae, a well-known model for genetic regulation. Despite this pathway being relative simple and well studied, various questions remain about the details of its regulatory mechanism. In the first part, I describe methodology for acquiring dynamic protein expression data. I use the green fluorescent protein (GFP) as a reporter and make measurements from cell populations growing in a microplate reader. Of particular interest is a technique I introduce for removing the autofluorescence that contaminates the GFP fluorescence signal. My technique makes it possible to detect expression even from very weakly expressed proteins. In the second part, I have developed a basic, deterministic model of the GAL pathway, which I then fit to dynamic protein expression data. I use the model to demonstrate that based on available data, the current understanding of the GAL pathway does capture a wide range of GAL system behaviours. I also identify and explore the GAL network's sensitivity to the relative levels of inducer and repressor proteins and discuss future directions for experiments and model development. In the third part, I present theoretical work that addresses the challenge of measuring protein-protein interactions in vivo from Förster resonance energy transfer (FRET) fluorescence data. I designed a computational tool that uses a Bayesian statistical framework to infer the dissociation constant and FRET efficiency from FRET data. One key advantage of this approach is that it produces probability distributions for these parameters of interest, revealing the uncertainty in the estimates obtained. I also demonstrate the experimental conditions, such as variations in levels of FRET donors and acceptors, that are necessary for the inference of the dissociation constant and FRET efficiency to be possible.
Au cours des deux dernières décennies, la biologie est passée d'une science réductionniste fondée sur l'observation qualitative à une science quantitative traitant les phénomènes biologiques comme des systèmes. Le développement de gènes rapporteurs fluorescents, d'instruments expérimentaux sophistiqués et de la puissance de calcul des ordinateurs ont été des éléments garants de cette transformation. Plus particulièrement, un élément clé de cette transformation a été l'utilisation croissante de modèles mathématiques pour explorer et vérifier notre compréhension de mécanismes biologiques complexes. Cette thèse traite de trois aspects de la modélisation des systèmes: l'acquisition de données, la création et l'évaluation d'un modèle et la mesure des valeurs de ses paramètres.Le système étudié dans les deux premières parties de cette thèse est la voie métabolique du galactose (GAL) chez la levure Saccharomyces cerevisiae. Cette voie est un exemple bien connu de régulation génétique. Bien qu'elle soit relativement simple et déjà bien étudiée, plusieurs questions subsistent quant aux détails reliés à son mécanisme de régulation. Dans la première partie, je décris la méthodologie pour l'acquisition de données dynamiques d'expression de protéines. J'utilise la protéine fluorescente verte (GFP) comme rapporteur et effectue des mesures sur des populations de cellules croissant dans un lecteur de microplaques. Notamment, je présente une technique permettant d'éliminer l'autofluorescence qui contamine le signal de fluorescence de la GFP. Cette technique permet même la détection de l'expression de protéines très faiblement exprimées. Dans la deuxième partie, je développe un modèle déterministe de base de la voie GAL, que je raccorde par la suite à des données dynamiques d'expression de protéines. J'utilise ce modèle pour démontrer qu'en se basant sur les données disponibles la compréhension théorique actuelle de la voie GAL capture une grande partie des comportements réels du système. Je discute aussi de possibilités pour l'élaboration subséquente du modèle. Dans la troisième partie, je présente un travail théorique qui traite de la difficulté de mesurer in vivo les interactions entre protéines à partir de données fluorescentes résultant du transfert d'énergie par résonance de type Förster (FRET). Je conçois un outil de calcul qui utilise les statistiques bayésiennes pour déduire la constante de dissociation et l'efficacité du FRET de données FRET. Un avantage clé de cette approche est qu'elle produit des distributions de probabilité pour ces paramètres d'intérêt, révélant l'incertitude des estimations obtenues. Je démontre aussi les conditions expérimentales, telles que les variations dans les concentrations de donneurs et d'accepteurs du FRET, requises pour permettre l'inférence de la constante de dissociation et de l'efficacité du FRET.
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Wang, You. "Development of yeast-based methods to screen for plant cytokinin-binding proteins." Access electronically, 2004. http://www.library.uow.edu.au/adt-NWU/public/adt-NWU20060123.141512/index.html.

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Petosa, Adamo. "Isolation of human scFv expressing cells from a yeast library using magnetic and fluorescence activated cell sorting." Thesis, McGill University, 2006. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=101733.

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The effective and efficient generation of both antibodies and antibody fragments to proteins of interest is vital, as antibodies and antibody fragments are required for an ever-increasing variety of therapeutic, diagnostic and analytical applications. The single chain variable fragment (scFv) is an antibody fragment consisting of a heavy chain variable region (VH) and a light chain variable region (VL) joined together by a flexible polypeptide linker. In 2003, Feldhaus et al. developed a nonimmune human scFv surface display library, in Saccharomyces cerevisiae , containing 109 different scFvs. Cells in the library expressing scFvs of interest can be isolated using magnetic cell sorting (MCS) and fluorescence activated cell sorting (FACS).
The reduced size of the scFv relative to the intact IgG allows it to penetrate tissue with greater ease and therefore reach epitopes within both tissue and cells that would otherwise remain inaccessible. As a result, one possible scFv application is the study of cartilage destruction by proteases that occurs in both normal joint development and arthritis. Antibody fragments would allow for cartilage degradative processes to be studied in vivo . Fluorescently tagged scFvs could penetrate intact cartilage tissue, bind to epitopes and then be localized using techniques such as dual photon confocal microscopy. This would not be possible using IgG molecules.
The yeast library developed by Feldhaus et al. was obtained for the potential isolation of cells expressing scFvs to cartilage neoepitopes. While found to possess an inherent Candida parapsilosis contamination, the surface display library was screened using three peptide-ovalbumin-biotin complexes. Peptides corresponding to observed cartilage neoepitopes were bound to biotinylated ovalbumin and added to the library for screening. Excess unlabelled ovalbumin was also added to the library to prevent the isolation of ovalbumin binding cells.
In all, two rounds of MCS and two rounds of FACS with all three antigens were used to screen the library for binders. A portion of the remaining library cells was then screened by MCS with a single antigen and eight individual clones were isolated. The affinity of these clones was determined and the scFv region of one clone was sequenced. Despite preventative measures, all eight clones isolated and analyzed were found to have an affinity to undetermined ovalbumin complex regions other than the peptides of interest. Still, cells expressing scFvs binding to a portion of the antigen complexes presented to the library were clearly enriched and subsequently isolated using MCS and FACS.
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He, Susu. "Functional localization study of acid trehalase (Ath1) and its secretion mechanism in the yeast Saccharomyces cerevisiae." Toulouse, INSA, 2009. http://eprint.insa-toulouse.fr/archive/00000334/.

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Des études précédentes sur la levure Saccharomyces cerevisiae ont proposé une vacuolaire localisation pour Ath1, qui est difficile à concilier avec sa capacité à hydrolyser tréhalose exogènes. Dans notre étude, nous avons utilisé la microscopie fluorescente à montrer que Ath1 est bien localisées dans la vacuole, mais aussi à la surface cellulaire. Néanmoins, que Ath1 à la surface cellulaire est responsable pour la croissance sur tréhalose, et Ath1 dans la vacuole est enclin à protéolyse. Deuxième partie sur l’étude de domaine protéique, nous avons montré que les N-terminales 131 acides aminés de Ath1 sont le domaine potentiel pour l’adressage à la surface, parmi le domaine transmembranaire est indispensable. Enfin, la voie de ciblage de Ath1 dans la cellule de levure a été étudié. En utilisant différent mutants impliqués dans la voie de ciblage à la vacuole, nous avons prouvé que le ciblage d'Ath1 vers la vacuole emprunte une voie intracellulaire, indépendant de l’endocytosis. De plus, le ciblage à la surface probablement emprunte la voie "classique" de sécrétion en aide d’un group de Sec protéines. Ces études sont en cours
Trehalose (alpha-D-glucopyranosyl (1→1) alpha-D-gluocopyranoside) is a non-reducing disaccharide found in many organisms including yeasts, fungi, bacteria, plants and insects. In the yeast Saccharomyces cerevisiae, trehalose is one of the major storage carbohydrates, accounting for more than 25% of cell dry mass depending on growth conditions and stage of the yeast life cycle (Hottiger et al. , 1987a; Jules et al. , 2008; Lillie and Pringle, 1980). The accumulation of intracellular trehalose has two potential functions. First, it constitutes an endogenous storage of carbon and energy during spore germination and in resting cells. Second, trehalose acts as a stabilizer of cellular membranes and proteins (Francois and Parrou, 2001; Simola et al. , 2000; Singer and Lindquist, 1998). In the yeast S. Cerevisiae, trehalose is hydrolyzed into glucose by the action of two types of trehalases: the ‘neutral trehalases’ encoded by NTH1 and NTH2 (Jules et al. , 2008; Mittenbuhler and Holzer, 1988), which are optimally active at pH 7, and the ‘acid trehalase’ encoded by ATH1, showing optimal activity at pH 4. 5 (Destruelle et al. , 1995). Neutral trehalase has been well studied and is known to hydrolyze trehalose in the cytosol. While fungal acid trehalases, including the yeast Candida albicans (Pedreno et al. , 2004) and Kluyveromyces lactis (Swaim et al. , 2008) enzymes, have been reported to be localized at the cell surface, the localization of the S. Cerevisiae acid trehalase is still a matter of controversy. In 1982, Wiemken and coworkers (Keller et al. , 1982) first identified this protein in vacuole-enriched fraction obtained by density gradient centrifugation of a yeast protoplast preparation. Vacuolar localization of acid trehalase was very recently supported by in vivo imaging analyses using GFP-Ath1 fusion constructs under the strong and constitutive TPI1 promoter (Huang et al. , 2007). Furthermore, these authors employed various trafficking mutants to show that this acid trehalase reaches its vacuolar destination through the multivesicular body (MVB) pathway. However, this localization contrasts with the fact that this enzyme allows yeast to grow on exogenous trehalose (Nwaka et al. , 1995b), and with a measurable Ath1 activity at the cell surface (Jules et al. , 2004)
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Assawajaruwan, Supasuda [Verfasser], and Bernd [Akademischer Betreuer] Hitzmann. "Development of an on-line process monitoring for yeast cultivations via 2D-fluorescence spectroscopy / Supasuda Assawajaruwan ; Betreuer: Bernd Hitzmann." Hohenheim : Kommunikations-, Informations- und Medienzentrum der Universität Hohenheim, 2019. http://d-nb.info/1177881446/34.

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Groß, Annett. "Genetically Tailored Yeast Strains for Cell-based Biosensors in White Biotechnology." Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2017. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-83341.

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This work was performed in the framework of two application-oriented research projects that focus on the generation and evaluation of fluorescent Saccharomyces (S.) cerevisiae-based sensor and reporter cells for white biotechnology as well as the extension of the conventional single-cell/single-construct principle of ordinary yeast biosensor approaches. Numerous products are currently generated by biotechnological processes which require continuous and precise process control and monitoring. These demands are only partially met by physical or physiochemical sensors since they measure parameters off-line or use surrogate parameters that consequently provide only indirect information about the actual process performance. Biosensors, in particular whole cell-based biosensors, have the unique potential to near-line and long-term monitor parameters such as nutrient availability during fermentation processes. Moreover, they allow for the assessment of an analyte’s biological relevance. Prototype yeast sensor and reporter strains derived from common laboratory strains were transformed with multicopy expression plasmids that mediate constitutive or inducible expression of a fluorescence reporter gene. Performance of these cells was examined by various qualitative and quantitative detection methods – representative of putative transducer technologies. Analyses were performed on the population level by microplate reader-based fluorometry and Western blot as well as on the single-cell level by fluorescence microscopy and flow cytometry. ‘Signature’ promoters that are activated or repressed during particular nutrient-limited growth conditions were selected in order to generate yeast nutrient sensor strains for monitoring the biological availability of nitrogen, phosphorus or sulphur. For each category, at least one promoter mediating at least threefold changed green fluorescence levels between sensor cells in non-limited and nutrient-limited conditions was identified. Sensor strains were evaluated in detail regarding sensitivity, analyte selectivity and the ability to restore basic fluorescence after shift from nutrient-limited to non-limited conditions (regeneration). The applicability for bioprocess monitoring purposes was tested by growth of yeast nutrient sensor cells in microalgae media and supernatants. Despite successful proof of principle, numerous challenges still need to be solved to realise prospective implementation in this field of white biotechnology. The major drawback of plasmid-borne detection constructs is a high fluorescence variance between individual cells. By generation of a nitrogen sensor strain with a genome-integrated detection construct, uniform expression on the single-cell level and simultaneous maintenance of basic properties (ability of fluorescence induction/regeneration and lack of cross-reactivity) was achieved. However, due to the singular detection construct per cell, significantly weaker overall fluorescence was observed. The traditional single-cell/single-construct approach was expanded upon in two ways. Firstly, a practical dual-colour sensor strain was created by simultaneous, constitutive expression of a red fluorescence reporter gene in green fluorescent nitrogen sensor cells. Secondly, an innovative cellular communication and signal amplification system inspired by the natural S. cerevisiae pheromone system and mating response was established successfully. It features the yeast pheromone alpha-factor as a trigger and alpha-factor-responsive reporter cells which express a fluorescence reporter gene from the pheromone-inducible FIG1 promoter as an output signal. The system was functional both with synthetic and cell-secreted alpha-factor, provided that recombinant cells were deleted for the alpha-factor protease Bar1p. Integration of amplifier cells which secrete alpha-factor in response to stimulation with the pheromone itself could increase the system\'s sensitivity further. Signal amplification was demonstrated for phosphorus sensor cells as a proof of concept. Therefore, the alpha-factor-based cellular communication and signal amplification system might be useful in applications that suffer from poor signal yield. Due to its modular design, the system could be applied in basically any cell-based biosensor or sensor-actor system. Immobilisation of the generated sensor and reporter cells in transparent natural polymers can be beneficial considering biosensor fabrication. Functionality of sensor and reporter cells in calcium-alginate beads or nano-printed arrays was successfully demonstrated. For the latter setup, fluorescence scanning and software-assisted fluorescence quantification was applied as a new detection method. In an experiment using an agarose-based two-compartment setup proposed by Jahn, 2011, properties of the alpha-factor-based cellular communication and signal amplification system after immobilisation were tested. These studies provide an initial experimental basis for an appropriate geometry of miniaturised immobilisation matrices with fluorescent yeast sensor and reporter cells in prospective biosensor designs.
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Korkmaz, Nuriye. "Self-assembly and Structure Investigation of Recombinant S-layer Proteins Expressed in Yeast for Nanobiotechnological Applications." Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2011. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-64317.

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In numerous Gram-negative and Gram-positive bacteria as well as in Archaea SL proteins form the outermost layer of the cell envelope. SL (glyco)monomers self-assemble with oblique (p2), tetragonal (p4), or hexagonal (p3, p6) symmetries [12]. SL subunits interact with each other and with the underlying cell surface by relatively weak non-covalent forces such as hydrogen-bonds, ionic bonds, salt-bridges or hydrophobic interactions. This makes them easy to isolate by applying chaotropic agents like urea and guanidine hydrochloride (GuHCl), chelating chemicals, or by changing the pH of the environment [10]. Upon dialysis in an ambient buffer monomers recrystallize into regular arrays that possess the forms of flat sheets, open ended cylinders, or spheres on solid substrates, at air-water intefaces and on lipid films, making them appealing for nanobiotechnological applications [3, 18]. The aim of this study was to investigate the structure, thermal stability, in vivo self-assembly process, recrystallization and metallization of three different recombinant SL proteins (SslA-eGFP, mSbsC-eGFP and S13240-eGFP) expressed in yeast S. cerevisiae BY4741 which could be further used in nanobiotechnological applications. In order to fulfill this aim, I investigated the in vivo expression of SL proteins (SslA, SbsC, S13240) tagged with eGFP (SL-eGFP) in the yeast S. cerevisiae BY4141. First, I characterized the heterologous expression of SL fusion constructs with growth and fluorescence measurements combined with Western blot analyses. Fluorescence microscopy investigations of overnight grown cultures showed that SslA-eGFP fusion protein was expressed as fluorescent patches, mSbsC-eGFP as tubular networks, and S13240-eGFP as hollow-like fibrillar network structures, while eGFP did not show any distinct structure Thermal stability of in vivo expressed SL-eGFP fusion proteins were investigated by fluorescence microscopy and immunodetection. In vivo self-assembly kinetics during mitosis and meiosis was the second main issue. In parallel, association of in vivo mSbsC-eGFP structures with the cellular components was of interest. A network of tubular structures in the cytosol of the transformed yeast cells that did not colocalize with microtubules or the actin cytoskeleton was observed. Time-resolved analysis of the formation of these structures during vegetative growth and sporulation was investigated by live fluorescence microscopy. While in meiosis ascospores seemed to receive assembled structures from the diploid cells, during mitosis surface layer structures were formed de novo in the buds. Surface layer assembly always started with the appearance of a dot-like structure in the cytoplasm, suggesting a single nucleation point. In order to get these in vivo SL assemblies stably outside the cells (in situ), cell distruption experiments were conducted. The tubular structures formed by the protein in vivo were retained upon bursting the cells by osmotic shock; however their average length was decreased. During dialysis, monomers obtained by treatment with chaotropic agents recrystallized again to form tube-like structures. This process was strictly dependent on calcium ions, with an optimal concentration of 10 mM. Further increase of the Ca2+ concentration resulted in multiple non-productive nucleation points. It was further shown that the lengths of the S-layer assemblies increased with time and could be controlled by pH. After 48 hours the average length at pH 9.0 was 4.13 µm compared to 2.69 µm at pH 5.5. Successful chemical deposition of platinum indicates the potential of recrystallized mSbsC-eGFP structures for nanobiotechnological applications. For example, such metalized protein nanotubes could be used in conductive nanocircuit technologies as nanowires.
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Sjöstrand, Linda. "Method Development for Thermal Stability Analysis by Circular Dichroism : Application to the Abp1p SH3 domain from yeast." Thesis, Linköpings universitet, Kemi, 2018. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-148286.

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Thermal stability is an important and interesting physical property of proteins. A common method to study it by is circular dichroism (CD) spectroscopy. The aim of this study was to test methods to improve thermal stability analysis by CD spectroscopy. Experiments were performed using the Abp1p SH3 domain from yeast as a model protein. Thermal denaturation was monitored at multiple wavelengths. It was concluded that for data sets of reasonable quality the choice of wavelength does not affect the results. An approach to estimate stability of thermophilic proteins was tested where thermal stability was measured at different concentrations of the denaturant GuHCl. The thermochemical data was used to estimate the stability in absence of GuHCl by extrapolation. The results were compared to those obtained from CD spectroscopy and differential scanning calorimetry. It was found that a stabilizing effect from low concentrations of GuHCl complicated the extrapolation. It is likely that this method is more successful if there is no stabilizing effect. The effect of ΔCp in stability parameter calculations was investigated with an experimentally and theoretically determined ΔCp. This was further investigated with synthetic data sets. The ΔCp used in calculations had no notable effect, as long as there was no cold denaturation. Although ΔCp is not necessary in calculations, it is an interesting parameter itself. ΔCp can be calculated from the thermochemical data used for extrapolation. The results in this study demonstrate robustness in thermal stability analysis by CD spectroscopy and a potential for development.
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Books on the topic "Yeast fluorescence"

1

Grigson, C. M. D. A microcontroller based flurometer for the detection of green fluorescent protein in genetically altered yeast. Manchester: UMIST, 1996.

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Book chapters on the topic "Yeast fluorescence"

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Denksteinová, B., D. Gášková, P. Heřman, J. Večeř, K. Sigler, J. Plášek, and J. Malínský. "Speed of Accumulation of the Membrane Potential Indicator diS-C3(3) in Yeast Cells." In Fluorescence Microscopy and Fluorescent Probes, 151–55. Boston, MA: Springer US, 1996. http://dx.doi.org/10.1007/978-1-4899-1866-6_21.

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Swayne, Theresa C., Anna C. Gay, and Liza A. Pon. "Fluorescence Imaging of Mitochondria in Yeast." In Methods in Molecular Biology, 433–59. Totowa, NJ: Humana Press, 2007. http://dx.doi.org/10.1007/978-1-59745-365-3_31.

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Learmonth, Robert P. "Membrane Fluidity in Yeast Adaptation: Insights from Fluorescence Spectroscopy and Microscopy." In Reviews in Fluorescence 2010, 67–93. New York, NY: Springer New York, 2011. http://dx.doi.org/10.1007/978-1-4419-9828-6_4.

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Kumar, Arun, and Manuel Mendoza. "Time-Lapse Fluorescence Microscopy of Budding Yeast Cells." In Methods in Molecular Biology, 1–8. New York, NY: Springer New York, 2016. http://dx.doi.org/10.1007/978-1-4939-3145-3_1.

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Scherthan, Harry. "FISH Targeting of Chromosomes and Subchromosomal Regions in Yeast." In Fluorescence In Situ Hybridization (FISH) — Application Guide, 347–59. Berlin, Heidelberg: Springer Berlin Heidelberg, 2009. http://dx.doi.org/10.1007/978-3-540-70581-9_30.

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Tramier, M., O. Holub, J. C. Croney, T. Ishi, S. E. Seifried, and D. M. Jameson. "Binding of Ethidium to Yeast tRNAPhe: A New Perspective on an Old Bromide." In Fluorescence Spectroscopy, Imaging and Probes, 111–21. Berlin, Heidelberg: Springer Berlin Heidelberg, 2002. http://dx.doi.org/10.1007/978-3-642-56067-5_6.

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Skarp, Kari-Pekka, Xueqiang Zhao, Marion Weber, and Jussi Jäntti. "Use of Bimolecular Fluorescence Complementation in Yeast Saccharomyces cerevisiae." In Membrane Trafficking, 165–75. Totowa, NJ: Humana Press, 2008. http://dx.doi.org/10.1007/978-1-59745-261-8_12.

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Learmonth, R. P., and E. Gratton. "Assessment of Membrane Fluidity in Individual Yeast Cells by Laurdan Generalised Polarisation and Multi-photon Scanning Fluorescence Microscopy." In Fluorescence Spectroscopy, Imaging and Probes, 241–52. Berlin, Heidelberg: Springer Berlin Heidelberg, 2002. http://dx.doi.org/10.1007/978-3-642-56067-5_14.

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Nagumo, Sachiyo, and Koji Okamoto. "Investigation of Yeast Mitophagy with Fluorescence Microscopy and Western Blotting." In Mitophagy, 71–83. New York, NY: Springer New York, 2017. http://dx.doi.org/10.1007/7651_2017_11.

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Lipatova, Zhanna, Jane J. Kim, and Nava Segev. "Ypt1 and TRAPP Interactions: Optimization of Multicolor Bimolecular Fluorescence Complementation in Yeast." In Methods in Molecular Biology, 107–16. New York, NY: Springer New York, 2015. http://dx.doi.org/10.1007/978-1-4939-2569-8_9.

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Conference papers on the topic "Yeast fluorescence"

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Bhatta, H., E. M. Goldys, and J. Ma. "Fluorescence and fluorescence-lifetime imaging microscopy (FLIM) to characterize yeast strains by autofluorescence." In Biomedical Optics 2006, edited by Daniel L. Farkas, Dan V. Nicolau, and Robert C. Leif. SPIE, 2006. http://dx.doi.org/10.1117/12.645354.

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PUCHKOV, EVGENY. "Subcellular Level Resolution Fluorescence Measurements in Yeast Cells by Image Analysis." In Eighth International Conference on Advances in Applied Science and Environmental Engineering - ASEE 2018. Institute of Research Engineers and Doctors, 2018. http://dx.doi.org/10.15224/978-1-63248-143-6-01.

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Wood, Christopher, Joseph Huff, Will Marshall, Elden Qingfeng Yu, Jay Unruh, Brian Slaughter, and Winfried Wiegraebe. "Fluorescence correlation spectroscopy as tool for high-content-screening in yeast (HCS-FCS)." In SPIE BiOS, edited by Jörg Enderlein, Zygmunt K. Gryczynski, and Rainer Erdmann. SPIE, 2011. http://dx.doi.org/10.1117/12.873947.

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"IMAGE ANALYSIS COMBINED FLUORESCENCE MICROSCOPY - Examples of ImageJ Software Application in Yeast Studies." In International Conference on Bioinformatics Models, Methods and Algorithms. SciTePress - Science and and Technology Publications, 2011. http://dx.doi.org/10.5220/0003120902580261.

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Chandrasekar, S. N., Abhishek Rao G, Sai Muthukumar V, Venketesh S, and R. Raghunatha Sarma. "Detection of Hotspots in Fluorescence Imaging of Yeast Cell Model used in Neuro-Degenerative Research." In 2021 International Conference on Advances in Electrical, Computing, Communication and Sustainable Technologies (ICAECT). IEEE, 2021. http://dx.doi.org/10.1109/icaect49130.2021.9392397.

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Mojica-Benavides, Martin, Amin A. Banaeiyan, David D. van Niekerk, Jacky L. Snoep, Anna-Karin Gustavsson, Caroline B. Adiels, and Mattias Goksör. "An optical tweezers, epi-fluorescence and microfluidic-setup for synchronization studies of glycolytic oscillations in living yeast cells." In SPIE Nanoscience + Engineering, edited by Kishan Dholakia and Gabriel C. Spalding. SPIE, 2016. http://dx.doi.org/10.1117/12.2236208.

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Aydin, Ali Selman, Abhinandan Dubey, Daniel Dovrat, Amir Aharoni, and Roy Shilkrot. "CNN Based Yeast Cell Segmentation in Multi-modal Fluorescent Microscopy Data." In 2017 IEEE Conference on Computer Vision and Pattern Recognition Workshops (CVPRW). IEEE, 2017. http://dx.doi.org/10.1109/cvprw.2017.105.

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Kato, Ilka T., Camila C. Santos, Endi Benetti, Denise P. L. A. Tenório, Paulo E. Cabral Filho, Caetano P. Sabino, Adriana Fontes, Beate S. Santos, Renato A. Prates, and Martha S. Ribeiro. "CdTe/CdS-MPA quantum dots as fluorescent probes to label yeast cells: synthesis, characterization and conjugation with Concanavalin A." In SPIE BiOS, edited by Wolfgang J. Parak, Kenji Yamamoto, and Marek Osinski. SPIE, 2012. http://dx.doi.org/10.1117/12.909060.

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