Gotowe bibliografie tematyczne / Live Cell Imaging Biosensors

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Gotowa bibliografia na temat „Live Cell Imaging Biosensors”

Autor: Grafiati
Data publikacji: 6 września 2023
Data aktualizacji: 7 września 2023

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Artykuły w czasopismach na temat "Live Cell Imaging Biosensors"

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Yoon, Sangpil, Yijia Pan, Kirk Shung i Yingxiao Wang. "FRET-Based Ca2+ Biosensor Single Cell Imaging Interrogated by High-Frequency Ultrasound". Sensors 20, nr 17 (3.09.2020): 4998. http://dx.doi.org/10.3390/s20174998.

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Fluorescence resonance energy transfer (FRET)-based biosensors have advanced live cell imaging by dynamically visualizing molecular events with high temporal resolution. FRET-based biosensors with spectrally distinct fluorophore pairs provide clear contrast between cells during dual FRET live cell imaging. Here, we have developed a new FRET-based Ca2+ biosensor using EGFP and FusionRed fluorophores (FRET-GFPRed). Using different filter settings, the developed biosensor can be differentiated from a typical FRET-based Ca2+ biosensor with ECFP and YPet (YC3.6 FRET Ca2+ biosensor, FRET-CFPYPet). A high-frequency ultrasound (HFU) with a carrier frequency of 150 MHz can target a subcellular region due to its tight focus smaller than 10 µm. Therefore, HFU offers a new single cell stimulations approach for FRET live cell imaging with precise spatial resolution and repeated stimulation for longitudinal studies. Furthermore, the single cell level intracellular delivery of a desired FRET-based biosensor into target cells using HFU enables us to perform dual FRET imaging of a cell pair. We show that a cell pair is defined by sequential intracellular delivery of the developed FRET-GFPRed and FRET-CFPYPet into two target cells using HFU. We demonstrate that a FRET-GFPRed exhibits consistent 10–15% FRET response under typical ionomycin stimulation as well as under a new stimulation strategy with HFU.
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Snell, Nicole, Vishnu Rao, Kendra Seckinger, Junyi Liang, Jenna Leser, Allison Mancini i M. Rizzo. "Homotransfer FRET Reporters for Live Cell Imaging". Biosensors 8, nr 4 (11.10.2018): 89. http://dx.doi.org/10.3390/bios8040089.

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Förster resonance energy transfer (FRET) between fluorophores of the same species was recognized in the early to mid-1900s, well before modern heterotransfer applications. Recently, homotransfer FRET principles have re-emerged in biosensors that incorporate genetically encoded fluorescent proteins. Homotransfer offers distinct advantages over the standard heterotransfer FRET method, some of which are related to the use of fluorescence polarization microscopy to quantify FRET between two fluorophores of identical color. These include enhanced signal-to-noise, greater compatibility with other optical sensors and modulators, and new design strategies based upon the clustering or dimerization of singly-labeled sensors. Here, we discuss the theoretical basis for measuring homotransfer using polarization microscopy, procedures for data collection and processing, and we review the existing genetically-encoded homotransfer biosensors.
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Secilmis, Melike, Hamza Yusuf Altun, Johannes Pilic, Yusuf Ceyhun Erdogan, Zeynep Cokluk, Busra Nur Ata, Gulsah Sevimli i in. "A Co-Culture-Based Multiparametric Imaging Technique to Dissect Local H2O2 Signals with Targeted HyPer7". Biosensors 11, nr 9 (14.09.2021): 338. http://dx.doi.org/10.3390/bios11090338.

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Multispectral live-cell imaging is an informative approach that permits detecting biological processes simultaneously in the spatial and temporal domain by exploiting spectrally distinct biosensors. However, the combination of fluorescent biosensors with distinct spectral properties such as different sensitivities, and dynamic ranges can undermine accurate co-imaging of the same analyte in different subcellular locales. We advanced a single-color multiparametric imaging method, which allows simultaneous detection of hydrogen peroxide (H2O2) in multiple cell locales (nucleus, cytosol, mitochondria) using the H2O2 biosensor HyPer7. Co-culturing of endothelial cells stably expressing differentially targeted HyPer7 biosensors paved the way for co-imaging compartmentalized H2O2 signals simultaneously in neighboring cells in a single experimental setup. We termed this approach COMPARE IT, which is an acronym for co-culture-based multiparametric imaging technique. Employing this approach, we detected lower H2O2 levels in mitochondria of endothelial cells compared to the cell nucleus and cytosol under basal conditions. Upon administering exogenous H2O2, the cytosolic and nuclear-targeted probes displayed similarly slow and moderate HyPer7 responses, whereas the mitochondria-targeted HyPer7 signal plateaued faster and reached higher amplitudes. Our results indicate striking differences in mitochondrial H2O2 accumulation of endothelial cells. Here, we present the method’s potential as a practicable and informative multiparametric live-cell imaging technique.
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Tiruthani, Karthik, Adam Mischler, Shoeb Ahmed, Jessica Mahinthakumar, Jason M. Haugh i Balaji M. Rao. "Design and evaluation of engineered protein biosensors for live-cell imaging of EGFR phosphorylation". Science Signaling 12, nr 584 (4.06.2019): eaap7584. http://dx.doi.org/10.1126/scisignal.aap7584.

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Live-cell fluorescence microscopy is broadly applied to study the dynamics of receptor-mediated cell signaling, but the availability of intracellular biosensors is limited. A biosensor based on the tandem SH2 domains from phospholipase C–γ1 (PLCγ1), tSH2-WT, has been used to measure phosphorylation of the epidermal growth factor receptor (EGFR). Here, we found that tSH2-WT lacked specificity for phosphorylated EGFR, consistent with the known promiscuity of SH2 domains. Further, EGF-stimulated membrane recruitment of tSH2-WT differed qualitatively from the expected kinetics of EGFR phosphorylation. Analysis of a mathematical model suggested, and experiments confirmed, that the high avidity of tSH2-WT resulted in saturation of its target and interference with EGFR endocytosis. To overcome the apparent target specificity and saturation issues, we implemented two protein engineering strategies. In the first approach, we screened a combinatorial library generated by random mutagenesis of the C-terminal SH2 domain (cSH2) of PLCγ1 and isolated a mutant form (mSH2) with enhanced specificity for phosphorylated Tyr992 (pTyr992) of EGFR. A biosensor based on mSH2 closely reported the kinetics of EGFR phosphorylation but retained cross-reactivity similar to tSH2-WT. In the second approach, we isolated a pTyr992-binding protein (SPY992) from a combinatorial library generated by mutagenesis of the Sso7d protein scaffold. Compared to tSH2-WT and mSH2, SPY992 exhibited superior performance as a specific, moderate-affinity biosensor. We extended this approach to isolate a biosensor for EGFR pTyr1148 (SPY1148). This approach of integrating theoretical considerations with protein engineering strategies can be generalized to design and evaluate suitable biosensors for various phospho-specific targets.
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Houser, Mei CQ, Steven S. Hou, Florian Perrin, Yuliia Turchyna, Brian J. Bacskai, Oksana Berezovska i Masato Maesako. "A Novel NIR-FRET Biosensor for Reporting PS/γ-Secretase Activity in Live Cells". Sensors 20, nr 21 (22.10.2020): 5980. http://dx.doi.org/10.3390/s20215980.

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Presenilin (PS)/γ-secretase plays a pivotal role in essential cellular events via proteolytic processing of transmembrane proteins that include APP and Notch receptors. However, how PS/γ-secretase activity is spatiotemporally regulated by other molecular and cellular factors and how the changes in PS/γ-secretase activity influence signaling pathways in live cells are poorly understood. These questions could be addressed by engineering a new tool that enables multiplexed imaging of PS/γ-secretase activity and additional cellular events in real-time. Here, we report the development of a near-infrared (NIR) FRET-based PS/γ-secretase biosensor, C99 720-670 probe, which incorporates an immediate PS/γ-secretase substrate APP C99 with miRFP670 and miRFP720 as the donor and acceptor fluorescent proteins, respectively. Extensive validation demonstrates that the C99 720-670 biosensor enables quantitative monitoring of endogenous PS/γ-secretase activity on a cell-by-cell basis in live cells (720/670 ratio: 2.47 ± 0.66 (vehicle) vs. 3.02 ± 1.17 (DAPT), ** p < 0.01). Importantly, the C99 720-670 and the previously developed APP C99 YPet-Turquoise-GL (C99 Y-T) biosensors simultaneously report PS/γ-secretase activity. This evidences the compatibility of the C99 720-670 biosensor with cyan (CFP)-yellow fluorescent protein (YFP)-based FRET biosensors for reporting other essential cellular events. Multiplexed imaging using the novel NIR biosensor C99 720-670 would open a new avenue to better understand the regulation and consequences of changes in PS/γ-secretase activity.
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Woehler, Andrew. "Simultaneous Quantitative Live Cell Imaging of Multiple FRET-Based Biosensors". PLoS ONE 8, nr 4 (16.04.2013): e61096. http://dx.doi.org/10.1371/journal.pone.0061096.

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Vilchez Mercedes, Samuel A., Ian Eder, Mona Ahmed, Ninghao Zhu i Pak Kin Wong. "Optimizing locked nucleic acid modification in double-stranded biosensors for live single cell analysis". Analyst 147, nr 4 (2022): 722–33. http://dx.doi.org/10.1039/d1an01802g.

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Banerjee, Swayoma, Luis Rene Garcia i Wayne K. Versaw. "Quantitative Imaging of FRET-Based Biosensors for Cell- and Organelle-Specific Analyses in Plants". Microscopy and Microanalysis 22, nr 2 (16.02.2016): 300–310. http://dx.doi.org/10.1017/s143192761600012x.

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AbstractGenetically encoded Förster resonance energy transfer (FRET)-based biosensors have been used to report relative concentrations of ions and small molecules, as well as changes in protein conformation, posttranslational modifications, and protein–protein interactions. Changes in FRET are typically quantified through ratiometric analysis of fluorescence intensities. Here we describe methods to evaluate ratiometric imaging data acquired through confocal microscopy of a FRET-based inorganic phosphate biosensor in different cells and subcellular compartments of Arabidopsis thaliana. Linear regression was applied to donor, acceptor, and FRET-derived acceptor fluorescence intensities obtained from images of multiple plants to estimate FRET ratios and associated location-specific spectral correction factors with high precision. FRET/donor ratios provided a combination of high dynamic range and precision for this biosensor when applied to the cytosol of both root and leaf cells, but lower precision when this ratiometric method was applied to chloroplasts. We attribute this effect to quenching of donor fluorescence because high precision was achieved with FRET/acceptor ratios and thus is the preferred ratiometric method for this organelle. A ligand-insensitive biosensor was also used to distinguish nonspecific changes in FRET ratios. These studies provide a useful guide for conducting quantitative ratiometric studies in live plants that is applicable to any FRET-based biosensor.
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Dobrzyński, Maciej, Marc-Antoine Jacques i Olivier Pertz. "Mining single-cell time-series datasets with Time Course Inspector". Bioinformatics 36, nr 6 (14.11.2019): 1968–69. http://dx.doi.org/10.1093/bioinformatics/btz846.

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Abstract Summary Thanks to recent advances in live cell imaging of biosensors, microscopy experiments can generate thousands of single-cell time-series. To identify sub-populations with distinct temporal behaviours that correspond to different cell fates, we developed Time Course Inspector (TCI)—a unique tool written in R/Shiny to combine time-series analysis with clustering. With TCI it is convenient to inspect time-series, plot different data views and remove outliers. TCI facilitates interactive exploration of various hierarchical clustering and cluster validation methods. We showcase TCI by analysing a single-cell signalling time-series dataset acquired using a fluorescent biosensor. Availability and implementation https://github.com/pertzlab/shiny-timecourse-inspector. Supplementary information Supplementary data are available at Bioinformatics online.
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Valetdinova, Kamila R., Tuyana B. Malankhanova, Suren M. Zakian i Sergey P. Medvedev. "The Cutting Edge of Disease Modeling: Synergy of Induced Pluripotent Stem Cell Technology and Genetically Encoded Biosensors". Biomedicines 9, nr 8 (5.08.2021): 960. http://dx.doi.org/10.3390/biomedicines9080960.

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The development of cell models of human diseases based on induced pluripotent stem cells (iPSCs) and a cell therapy approach based on differentiated iPSC derivatives has provided a powerful stimulus in modern biomedical research development. Moreover, it led to the creation of personalized regenerative medicine. Due to this, in the last decade, the pathological mechanisms of many monogenic diseases at the cell level have been revealed, and clinical trials of various cell products derived from iPSCs have begun. However, it is necessary to reach a qualitatively new level of research with cell models of diseases based on iPSCs for more efficient searching and testing of drugs. Biosensor technology has a great application prospect together with iPSCs. Biosensors enable researchers to monitor ions, molecules, enzyme activities, and channel conformation in live cells and use them in live imaging and drug screening. These probes facilitate the measurement of steady-state concentrations or activity levels and the observation and quantification of in vivo flux and kinetics. Real-time monitoring of drug action in a specific cellular compartment, organ, or tissue type; the ability to screen at the single-cell resolution; and the elimination of the false-positive results caused by low drug bioavailability that is not detected by in vitro testing methods are a few of the benefits of using biosensors in drug screening. Here, we discuss the possibilities of using biosensor technology in combination with cell models based on human iPSCs and gene editing systems. Furthermore, we focus on the current achievements and problems of using these methods.
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Rozprawy doktorskie na temat "Live Cell Imaging Biosensors"

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Konishi, Yoshinobu. "Live-cell FRET imaging reveals a role of extracellular signal-regulated kinase activity dynamics in thymocyte motility". Kyoto University, 2019. http://hdl.handle.net/2433/242374.

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Hung, Yin Pun. "Single Cell Imaging of Metabolism with Fluorescent Biosensors". Thesis, Harvard University, 2012. http://dissertations.umi.com/gsas.harvard:10147.

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Cells utilize various signal transduction networks to regulate metabolism. Nevertheless, a quantitative understanding of the relationship between growth factor signaling and metabolic state at the single cell level has been lacking. The signal transduction and metabolic states could vary widely among individual cells. However, such cell-to-cell variation might be masked by the bulk measurements obtained from conventional biochemical methods. To assess the spatiotemporal dynamics of metabolism in individual intact cells, we developed genetically encoded biosensors based on fluorescent proteins. As a key redox cofactor in metabolism, NADH has been implicated in the Warburg effect, the abnormal metabolism of glucose that is a hallmark of cancer cells. To date, however, sensitive and specific detection of NADH in the cytosol of individual live cells has been difficult. We engineered a fluorescent biosensor of NADH by combining a circularly permuted green fluorescent protein variant with a bacterial NADH-binding protein Rex. The optimized biosensor Peredox reports cytosolic \(NADH:NAD^+\) ratios in individual live cells and can be calibrated with exogenous lactate and pyruvate. Notably pH resistant, this biosensor can be used in several cultured and primary cell types and in a high-content imaging format. We then examined the single cell dynamics of glycolysis and energy-sensing signaling pathways using Peredox and other fluorescent biosensors: AMPKAR, a sensor of the AMPK activity; and FOXO3-FP, a fluorescently-tagged protein domain from Forkhead transcription factor FOXO3 to report on the PI3K/Akt pathway activity. With perturbation to growth factor signaling, we observed a transient response in the cytosolic \(NADH:NAD^+\) redox state. In contrast, with partial inhibition of glycolysis by iodoacetate, individual cells varied substantially in their responses, and cytosolic \(NADH:NAD^+\) ratios oscillated between high and low states with a regular, approximately half-hour period, persisting for hours. These glycolytic NADH oscillations appeared to be cell-autonomous and coincided with the activation of the PI3K/Akt pathway but not the AMPK pathway. These results suggest a dynamic coupling between growth factor signaling and metabolic parameters. Overall, this thesis presents novel optical tools to assess metabolic dynamics – and to unravel the elaborate and complex integration of glucose metabolism and signaling pathways at the single cell level.
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Kosmacek, Elizabeth Anne Ianzini Fiorenza Mackey Michael A. "Live cell imaging technology development for cancer research". [Iowa City, Iowa] : University of Iowa, 2009. http://ir.uiowa.edu/etd/388.

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Chyan, Wen Ph D. Massachusetts Institute of Technology. "Fluorogenic probes for live-cell imaging of biomolecules". Thesis, Massachusetts Institute of Technology, 2018. http://hdl.handle.net/1721.1/118216.

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Thesis: Ph. D. in Biological Chemistry, Massachusetts Institute of Technology, Department of Chemistry, 2018.
Cataloged from PDF version of thesis.
Includes bibliographical references (pages 231-249).
Fluorogenic probes, small-molecule sensors that unmask brilliant fluorescence upon exposure to specific stimuli, are essential tools for chemical biology. Probes that detect enzymatic activity can be used to illuminate the complex dynamics of biological processes at a level of spatiotemporal detail and sensitivity unmatched by other techniques. This dissertation describes the development of new fluorophore chemistries to expand our current fluorogenic probe toolkit and the subsequent application of these probes to study dynamic cell transport processes. Chapter 1. Enzyme-Activated Fluorogenic Probes for Live-Cell and In Vivo Imaging. Chapter 1 reviews recent advances in enzyme-activated fluorogenic probes for biological imaging, organized by enzyme classification. This review surveys recent masking strategies, different modes of enzymatic activation, and the breadth of current and future probe applications. Key challenges, such as probe selectivity and spectroscopic requirements, are described in this chapter along with therapeutic and diagnostic opportunities that can be accessed by surmounting these challenges. Chapter 2. Electronic and Steric Optimization of Fluorogenic Probes for Biomolecular Imaging. In many fluorogenic probes, the intrinsic fluorescence of a small-molecule fluorophore is masked by ester masking groups until entry into a cell, where endogenous esterases catalyze the hydrolysis of esters, generating fluorescence. The susceptibility of masking groups to spontaneous hydrolysis is a major limitation of these probes. Previous attempts to address this problem have incorporated auto-immolative linkers at the cost of atom economy and synthetic adversity. In this chapter, I report on a linker-free strategy that employs adventitious electronic and steric interactions in easy-to-synthesize probes. I find that halogen-carbonyl n-->[pi]* interactions and acyl group size are optimized in 2',7'-dichlorofluorescein diisobutyrate. This probe is relatively stable to spontaneous hydrolysis but is a highly reactive substrate for esterases both in vitro and in cellulo, yielding a bright, photostable fluorogenic probe with utility in biomolecular imaging. Chapter 3. Cellular Uptake of Large Monofunctionalized Dextrans. Dextrans are a versatile class of polysaccharides with applications that span medicine, cell biology, food science, and consumer goods. In Chapter 3, I apply the electronically stabilized probe described in Chapter 2 to study the cellular uptake of a new type of large monofunctionalized dextran that exhibits unusual properties: efficient cytosolic and nuclear uptake. This dextran permeates various human cell types without the use of transfection agents, electroporation, or membrane perturbation. Cellular uptake occurs primarily through active transport via receptor-mediated processes. These monofunctionalized dextrans could serve as intracellular delivery platforms for drugs or other cargos. Chapter 4. Paired Nitroreductase-Probe System to Quantify the Cytosolic Delivery of Biomolecules. Cytosolic delivery of large biomolecules is a significant barrier to therapeutic applications of CRISPR, RNAi, and biologics such as proteins with anticancer properties. In Chapter 4, I describe a new paired enzyme-probe system to quantify cytosolic delivery of biomolecules-a valuable resource for elucidating mechanistic details and improving delivery of therapeutics. I designed and optimized a nitroreductase fusion protein that embeds in the cytosolic face of outer mitochondrial membranes, providing several key improvements over unanchored reporter enzymes. In parallel, I prepared and assessed a panel of nitroreductase-activated probes for favorable spectroscopic and enzymatic activation properties. Together, the nitroreductase fusion protein and fluorogenic probes provide a rapid, generalizable tool that is well-poised to quantify cytosolic delivery of biomolecules. Chapter 5. Future Directions. This chapter outlines several future directions for expanding the scope of fluorogenic probes and developing new biological applications. Additionally, Chapter 5 is followed by an appendix describing a tunable rhodol fluorophore scaffold for improved spectroscopic properties and versatility. Overall, the work described in this thesis illustrates the power of enzyme-activated fluorogenic probes to provide fresh insight into dynamic biological processes, with direct implications for improved therapeutic delivery.
by Wen Chyan.
Ph. D. in Biological Chemistry
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Büchele, Benjamin. "Live Cell Imaging des Hepatitis C Virus Replikationskomplexes". [S.l. : s.n.], 2004. http://nbn-resolving.de/urn:nbn:de:bsz:25-opus-59102.

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Kosmacek, Elizabeth Anne. "Live cell imaging technology development for cancer research". Diss., University of Iowa, 2009. https://ir.uiowa.edu/etd/388.

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Live cell imaging is a unique tool for cellular research with a wide variety of applications. By streaming digital microscopic images an investigator can observe the dynamic morphology of a cell, track cell movement on a surface, and measure quantities or localization patterns of fluorescently labeled proteins or molecules. Digital image sequences contain a vast amount of information in the form of visually detectable morphological changes in the cell. We designed computer programs that allow the manual identification of visible events in live cell digital image sequences [Davis et al. 2007]. Once identified, the data are analyzed using algorithms to calculate the yield of individual events per cell over the time course of image acquisition. The sequence of event data is also constructed into directed acyclic graphs and through the use of a subgraph isomorphism algorithm we are able to detect specified patterns of events originating from a single cell. Two projects in the field of cancer research are here discussed that describe and validate the application of the event analysis programs. In the first project, mitotic catastrophe (MC) research [Ianzini and Mackey, 1997; Ianzini and Mackey, 1998; reviewed by Ianzini and Mackey, 2007] is enhanced with the addition of live cell imaging to traditional laboratory experiments. The event analysis program is used to describe the yield of normal or abnormal divisions, fusions, and cell death, and to detect patterns of reductive division and depolyploidization in cells undergoing radiation-induced MC. Additionally, the biochemical and molecular data used in conjunction with live cell imaging data are presented to illustrate the usefulness of combining biology and engineering techniques to elucidate pathways involved in cell survival under different detrimental cell conditions. The results show that the timing of depolyploidization in MC cells correlates with increased multipolar divisions, up-regulation of meiosis-specific genes, and the production of mononucleated cell progeny. It was confirmed that mononucleated cells are produced from multipolar divisions and these cells are capable of resuming normal divisions [Ianzini et al., 2009]. The implications for the induction of meiosis as a mechanism of survival after radiation treatment are discussed. In the second project, the effects of long-term fluorescence excitation light exposure are examined through measurements of cell division and cell death. In the field of live cell imaging, probably the most modern and most widely utilized technique is fluorescence detection for intracellular organelles, proteins, and molecules. While the technologies required to label and detect fluorescent molecules in a cell are well developed, they are not idealized for long term measurements as both the probes and excitation light are toxic to the cells [Wang and Nixon, 1978; Bradley and Sharkey, 1977]. From the event analysis data it was determined that fluorescence excitation light is toxic to multiple cell lines observed as the reduction of normal cell division, induction of cell death, and apparent morphological aberrations.
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Caporale, Chiara. "Luminescent Iridium Tetrazolato Markers for Live Cell Imaging". Thesis, Curtin University, 2018. http://hdl.handle.net/20.500.11937/70386.

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In this research, a series of iridium(III) tetrazolato complexes were synthesised and their photophysical and biological properties investigated. Both the cyclometalated and the ancillary ligands were systematically modified by substitution of functional groups or by increasing the extension of the  conjugation. This approach allowed a more systematic rationalisation of the structure-activity relationship, highlighting how variations in the chemical structure and charge might influence the biological behaviour of these complexes.
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Danylchuk, Dmytro. "Environment-sensitive targeted fluorescent probes for live-cell imaging". Thesis, Strasbourg, 2021. http://www.theses.fr/2021STRAF012.

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Le ciblage, l'imagerie et le sondage spécifiques des membranes plasmiques et des organites intracellulaires peuvent être faits par des sondes fluorescentes à façon sensibles à la polarité. Ici, un nouveau fragment ciblant la membrane plasmique à été développé et testé dans cinq colorants cyanines, montrant d'excellentes performances en microscopie cellulaire et in vivo. Le fragment à été greffé à un fluorophore solvatochrome Prodan, donnant une sonde de membrane plasmique avec une sensibilité élevée à l'ordre lipidique. Le rouge de Nil, greffé aux fragments avec les chaînes alkyles C12 et C4, à donné deux sondes solvatochromes à membrane plasmique : NR12A pour la microscopie conventionnelle, et NR4A pour la microscopie à super-résolution PAINT. Le rouge de Nil avec des groupes ciblant les organites à donné un éventail de sondes sensibles à la polarité et à l'ordre lipidique dans les membranes des organites. Les sondes synthétisées trouveront des applications en bioimagerie, biologie cellulaire, biophysique ou mécanobiologie
Specific targeting, imaging and probing of cell plasma membranes and intracellular organelles can be addressed by rationally designed polarity-sensitive fluorescent probes. Here, a new efficient plasma membrane-targeting moiety was developed and tested in five cyanine dyes, showing excellent performance in cellular and in vivo microscopy. Next, the targeting moiety was grafted to a solvatochromic dye Prodan, yielding a plasma membrane probe with high lipid order sensitivity. Modifying a Nile Red using the moieties with varied alkyl chain lengths resulted in two solvatochromic plasma membrane probes: NR12A with high affinity to membranes for conventional microscopy, and NR4A, a low-affinity probe for PAINT super-resolution microscopy. Tethering Nile Red with organelle-targeted groups yielded an array of probes, able to sense polarity and lipid order in organelle membranes. The synthesized probes will find applications in bioimaging, cell biology, biophysics or mechanobiology
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Sörman, Paulsson Elsa. "Evaluation of In-Silico Labeling for Live Cell Imaging". Thesis, Umeå universitet, Institutionen för fysik, 2021. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-180590.

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Today new drugs are tested on cell cultures in wells to minimize time, cost, andanimal testing. The cells are studied using microscopy in different ways and fluorescentprobes are used to study finer details than the light microscopy can observe.This is an invasive method, so instead of molecular analysis, imaging can be used.In this project, phase-contrast microscopy images of cells together with fluorescentmicroscopy images were used. We use Machine Learning to predict the fluorescentimages from the light microscopy images using a strategy called In-Silico Labeling.A Convolutional Neural Network called U-Net was trained and showed good resultson two different datasets. Pixel-wise regression, pixel-wise classification, andimage classification with one cell in each image was tested. The image classificationwas the most difficult part due to difficulties assigning good quality labels tosingle cells. Pixel-wise regression showed the best result.
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Han, Hongqing. "Towards accurate and efficient live cell imaging data analysis". Doctoral thesis, Humboldt-Universität zu Berlin, 2021. http://dx.doi.org/10.18452/22324.

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Dynamische zelluläre Prozesse wie Zellzyklus, Signaltransduktion oder Transkription zu analysieren wird Live-cell-imaging mittels Zeitraffermikroskopie verwendet. Um nun aber Zellabstammungsbäume aus einem Zeitraffervideo zu extrahieren, müssen die Zellen segmentiert und verfolgt werden können. Besonders hier, wo lebende Zellen über einen langen Zeitraum betrachtet werden, sind Fehler in der Analyse fatal: Selbst eine extrem niedrige Fehlerrate kann sich amplifizieren, wenn viele Zeitpunkte aufgenommen werden, und damit den gesamten Datensatz unbrauchbar machen. In dieser Arbeit verwenden wir einen einfachen aber praktischen Ansatz, der die Vorzüge der manuellen und automatischen Ansätze kombiniert. Das von uns entwickelte Live-cell-Imaging Datenanalysetool ‘eDetect’ ergänzt die automatische Zellsegmentierung und -verfolgung durch Nachbearbeitung. Das Besondere an dieser Arbeit ist, dass sie mehrere interaktive Datenvisualisierungsmodule verwendet, um den Benutzer zu führen und zu unterstützen. Dies erlaubt den gesamten manuellen Eingriffsprozess zu rational und effizient zu gestalten. Insbesondere werden zwei Streudiagramme und eine Heatmap verwendet, um die Merkmale einzelner Zellen interaktiv zu visualisieren. Die Streudiagramme positionieren ähnliche Objekte in unmittelbarer Nähe. So kann eine große Gruppe ähnlicher Fehler mit wenigen Mausklicks erkannt und korrigiert werden, und damit die manuellen Eingriffe auf ein Minimum reduziert werden. Die Heatmap ist darauf ausgerichtet, alle übersehenen Fehler aufzudecken und den Benutzern dabei zu helfen, bei der Zellabstammungsrekonstruktion schrittweise die perfekte Genauigkeit zu erreichen. Die quantitative Auswertung zeigt, dass eDetect die Genauigkeit der Nachverfolgung innerhalb eines akzeptablen Zeitfensters erheblich verbessern kann. Beurteilt nach biologisch relevanten Metriken, übertrifft die Leistung von eDetect die derer Tools, die den Wettbewerb ‘Cell Tracking Challenge’ gewonnen haben.
Live cell imaging based on time-lapse microscopy has been used to study dynamic cellular behaviors, such as cell cycle, cell signaling and transcription. Extracting cell lineage trees out of a time-lapse video requires cell segmentation and cell tracking. For long term live cell imaging, data analysis errors are particularly fatal. Even an extremely low error rate could potentially be amplified by the large number of sampled time points and render the entire video useless. In this work, we adopt a straightforward but practical design that combines the merits of manual and automatic approaches. We present a live cell imaging data analysis tool `eDetect', which uses post-editing to complement automatic segmentation and tracking. What makes this work special is that eDetect employs multiple interactive data visualization modules to guide and assist users, making the error detection and correction procedure rational and efficient. Specifically, two scatter plots and a heat map are used to interactively visualize single cells' visual features. The scatter plots position similar results in close vicinity, making it easy to spot and correct a large group of similar errors with a few mouse clicks, minimizing repetitive human interventions. The heat map is aimed at exposing all overlooked errors and helping users progressively approach perfect accuracy in cell lineage reconstruction. Quantitative evaluation proves that eDetect is able to largely improve accuracy within an acceptable time frame, and its performance surpasses the winners of most tasks in the `Cell Tracking Challenge', as measured by biologically relevant metrics.
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Książki na temat "Live Cell Imaging Biosensors"

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Kim, Sung-Bae, red. Live Cell Imaging. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1258-3.

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Papkovsky, Dmitri B., red. Live Cell Imaging. Totowa, NJ: Humana Press, 2010. http://dx.doi.org/10.1007/978-1-60761-404-3.

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Live cell imaging: Methods and protocols. New York, NY: Humana Press, 2010.

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1939-, Goldman Robert D., Swedlow Jason i Spector David L, red. Live cell imaging: A laboratory manual. Wyd. 2. Cold Spring Harbor, N.Y: Cold Spring Harbor Laboratory Press, 2010.

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Live cell imaging: A laboratory manual. Wyd. 2. Cold Spring Harbor, N.Y: Cold Spring Harbor Laboratory Press, 2010.

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1939-, Goldman Robert D., Swedlow Jason i Spector David L, red. Live cell imaging: A laboratory manual. Wyd. 2. Cold Spring Harbor, N.Y: Cold Spring Harbor Laboratory Press, 2010.

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Masters, Barry R. Confocal microscopy and multiphoton excitation microscopy: The genesis of live cell imaging. Bellingham, WA: SPIE Press, 2006.

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Confocal microscopy and multiphoton excitation microscopy: The genesis of live cell imaging. Bellingham, Wash: SPIE Press, 2005.

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Tsukahara, Shinji. Kagaku busshitsu no yūgaisei hyōka no kōritsuka o mezashita aratana shinkei dokusei shikenhō no kaihatsu: Kankyōshō kankyō kenkyū gijutsu kaihatsu suishinhi shūryō kenkyū seika hōkokusho : Heisei 20-nendo--Heisei 21-nendo = Use of live cell imaging for efficient neurotoxicology methods : environment research and technology development fund. [Tokyo]: Kankyōshō Sōgō Kankyō Seisakukyoku Kankyō Hokenbu Kankyō Anzenka Kankyō Risuku Hyōkashitsu, 2010.

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Tsukahara, Shinji. Kagaku busshitsu no yūgaisei hyōka no kōritsuka o mezashita aratana shinkei dokusei shikenhō no kaihatsu: Kankyōshō kankyō kenkyū gijutsu kaihatsu suishinhi shūryō kenkyū seika hōkokusho : Heisei 20-nendo--Heisei 21-nendo = Use of live cell imaging for efficient neurotoxicology methods : environment research and technology development fund. [Tokyo]: Kankyōshō Sōgō Kankyō Seisakukyoku Kankyō Hokenbu Kankyō Anzenka Kankyō Risuku Hyōkashitsu, 2010.

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Części książek na temat "Live Cell Imaging Biosensors"

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Bravo-Cordero, Jose Javier, Yasmin Moshfegh, John Condeelis i Louis Hodgson. "Live Cell Imaging of RhoGTPase Biosensors in Tumor Cells". W Adhesion Protein Protocols, 359–70. Totowa, NJ: Humana Press, 2013. http://dx.doi.org/10.1007/978-1-62703-538-5_22.

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Sato, Moritoshi. "Genetically Encoded Fluorescent Biosensors for Live Cell Imaging of Lipid Dynamics". W Methods in Molecular Biology, 73–81. Totowa, NJ: Humana Press, 2013. http://dx.doi.org/10.1007/978-1-62703-622-1_6.

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Mishina, Natalia M., i Vsevolod V. Belousov. "Live-Cell STED Imaging with the HyPer2 Biosensor". W Methods in Molecular Biology, 21–28. New York, NY: Springer New York, 2017. http://dx.doi.org/10.1007/978-1-4939-7265-4_3.

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Kellenberger, Colleen A., Zachary F. Hallberg i Ming C. Hammond. "Live Cell Imaging Using Riboswitch-Spinach tRNA Fusions as Metabolite-Sensing Fluorescent Biosensors". W RNA Scaffolds, 87–103. New York, NY: Springer New York, 2015. http://dx.doi.org/10.1007/978-1-4939-2730-2_8.

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Ouyang, Mingxing, Shaoying Lu i Yingxiao Wang. "Genetically Encoded Fluorescent Biosensors for Live-Cell Imaging of MT1-MMP Protease Activity". W Methods in Molecular Biology, 163–74. Totowa, NJ: Humana Press, 2013. http://dx.doi.org/10.1007/978-1-62703-622-1_13.

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Manna, Sudeshna, Colleen A. Kellenberger, Zachary F. Hallberg i Ming C. Hammond. "Live Cell Imaging Using Riboswitch–Spinach tRNA Fusions as Metabolite-Sensing Fluorescent Biosensors". W RNA Scaffolds, 121–40. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1499-0_10.

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Hung, Yin Pun, i Gary Yellen. "Live-Cell Imaging of Cytosolic NADH–NAD+ Redox State Using a Genetically Encoded Fluorescent Biosensor". W Methods in Molecular Biology, 83–95. Totowa, NJ: Humana Press, 2013. http://dx.doi.org/10.1007/978-1-62703-622-1_7.

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Lopreside, Antonia, Maria Maddalena Calabretta, Laura Montali, Aldo Roda i Elisa Michelini. "Live Cell Immobilization". W Handbook of Cell Biosensors, 1–18. Cham: Springer International Publishing, 2019. http://dx.doi.org/10.1007/978-3-319-47405-2_146-1.

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Lopreside, Antonia, Maria Maddalena Calabretta, Laura Montali, Aldo Roda i Elisa Michelini. "Live Cell Immobilization". W Handbook of Cell Biosensors, 479–96. Cham: Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-23217-7_146.

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Van Kerckvoorde, Melinda, Matthew J. Ford, Patricia L. Yeyati, Pleasantine Mill i Richard L. Mort. "Live Imaging and Analysis of Cilia and Cell Cycle Dynamics with the Arl13bCerulean-Fucci2a Biosensor and Fucci Tools". W Methods in Molecular Biology, 291–309. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1538-6_21.

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Streszczenia konferencji na temat "Live Cell Imaging Biosensors"

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Pavlinska, Zuzana, Zuzana Kronekova, Igor Lacik, Dusana Trelova, Filip Razga, Veronika Nemethova, Lucia Uhelska, Alzbeta Marcek Chorvatova, Tibor Teplicky i Dusan Chorvat. "A bio-inspired design of live cell biosensors". W Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications XV, redaktorzy Alexander N. Cartwright, Dan V. Nicolau i Dror Fixler. SPIE, 2018. http://dx.doi.org/10.1117/12.2288789.

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Lu, Shaoying, i Yingxiao Wang. "Application of FRET biosensors and computational analysis for live cell imaging". W SPIE BiOS: Biomedical Optics, redaktorzy Alexander P. Savitsky i Yingxiao Wang. SPIE, 2009. http://dx.doi.org/10.1117/12.812183.

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Kamioka, Yuji, Kenta Sumiyama, Rei Mizuno i Michiyuki Matsuda. "Live imaging of transgenic mice expressing FRET biosensors". W 2013 35th Annual International Conference of the IEEE Engineering in Medicine and Biology Society (EMBC). IEEE, 2013. http://dx.doi.org/10.1109/embc.2013.6609453.

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Boero, Cristina, Maria Antonietta Casulli, Jacopo Olivo, Lorenzo Foglia, Sandro Carrara i Giovanni De Micheli. "Live demonstration: In-situ biosensors array for cell culture monitoring". W 2014 IEEE Biomedical Circuits and Systems Conference (BioCAS). IEEE, 2014. http://dx.doi.org/10.1109/biocas.2014.6981676.

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Moscelli, Nicola, Wojciech Witarski, Sander van den Driesche i Michael J. Vellekoop. "In incubator live cell imaging platform". W SPIE Microtechnologies, redaktorzy Ulrich Schmid, José Luis Sánchez-Rojas i Monika Leester-Schaedel. SPIE, 2011. http://dx.doi.org/10.1117/12.886908.

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Schneckenburger, Herbert, Verena Richter, Sarah Bruns, Thomas Bruns, Mathis Piper, Petra Weber, Michael Wagner i Christoph Cremer. "Axial tomography in 3D live cell microscopy". W Advances in Microscopic Imaging, redaktorzy Francesco S. Pavone, Emmanuel Beaurepaire i Peter T. So. SPIE, 2017. http://dx.doi.org/10.1117/12.2286602.

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Ren, Juan, i Qingze Zou. "Modeling of sample deformation in atomic force microscope imaging on live cell: Live mammalian cell imaging example?" W 2017 American Control Conference (ACC). IEEE, 2017. http://dx.doi.org/10.23919/acc.2017.7963048.

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Sim, J. Y., N. Borghi, A. Ribeiro, M. Sorokina, O. Shcherbakova, D. Ramallo, A. Dunn, W. J. Nelson i B. L. Pruitt. "Uniaxial cell stretcher enables high resolution live cell imaging". W 2012 IEEE 25th International Conference on Micro Electro Mechanical Systems (MEMS). IEEE, 2012. http://dx.doi.org/10.1109/memsys.2012.6170320.

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Mellors, Ben O., Hamid Dehghani, Christopher R. Howle i Abigail M. Spear. "Cell trauma detection using infra-red live cell imaging". W Preclinical and Clinical Optical Diagnostics, redaktorzy J. Quincy Brown i Ton G. van Leeuwen. SPIE, 2019. http://dx.doi.org/10.1117/12.2525012.

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Schneckenburger, H., V. Richter i C. Cremer. "Multi-modal Imaging in Live Cell Microscopy". W 2020 International Conference Laser Optics (ICLO). IEEE, 2020. http://dx.doi.org/10.1109/iclo48556.2020.9285496.

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Raporty organizacyjne na temat "Live Cell Imaging Biosensors"

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Ray, Judhajeet. Aptamer sensors for live-cell imaging of Pol II promoter activity. Office of Scientific and Technical Information (OSTI), grudzień 2014. http://dx.doi.org/10.2172/1227286.

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Zhang, Yun. Real time imaging of live cell ATP leaking or release events by chemiluminescence microscopy. Office of Scientific and Technical Information (OSTI), grudzień 2008. http://dx.doi.org/10.2172/964390.

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Belkin, Shimshon, Sylvia Daunert i Mona Wells. Whole-Cell Biosensor Panel for Agricultural Endocrine Disruptors. United States Department of Agriculture, grudzień 2010. http://dx.doi.org/10.32747/2010.7696542.bard.

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Objectives: The overall objective as defined in the approved proposal was the development of a whole-cell sensor panel for the detection of endocrine disruption activities of agriculturally relevant chemicals. To achieve this goal several specific objectives were outlined: (a) The development of new genetically engineered wholecell sensor strains; (b) the combination of multiple strains into a single sensor panel to effect multiple response modes; (c) development of a computerized algorithm to analyze the panel responses; (d) laboratory testing and calibration; (e) field testing. In the course of the project, mostly due to the change in the US partner, three modifications were introduced to the original objectives: (a) the scope of the project was expanded to include pharmaceuticals (with a focus on antibiotics) in addition to endocrine disrupting chemicals, (b) the computerized algorithm was not fully developed and (c) the field test was not carried out. Background: Chemical agents, such as pesticides applied at inappropriate levels, may compromise water quality or contaminate soils and hence threaten human populations. In recent years, two classes of compounds have been increasingly implicated as emerging risks in agriculturally-related pollution: endocrine disrupting compounds (EDCs) and pharmaceuticals. The latter group may reach the environment by the use of wastewater effluents, whereas many pesticides have been implicated as EDCs. Both groups pose a threat in proportion to their bioavailability, since that which is biounavailable or can be rendered so is a priori not a threat; bioavailability, in turn, is mediated by complex matrices such as soils. Genetically engineered biosensor bacteria hold great promise for sensing bioavailability because the sensor is a live soil- and water-compatible organism with biological response dynamics, and because its response can be genetically “tailored” to report on general toxicity, on bioavailability, and on the presence of specific classes of toxicants. In the present project we have developed a bacterial-based sensor panel incorporating multiple strains of genetically engineered biosensors for the purpose of detecting different types of biological effects. The overall objective as defined in the approved proposal was the development of a whole-cell sensor panel for the detection of endocrine disruption activities of agriculturally relevant chemicals. To achieve this goal several specific objectives were outlined: (a) The development of new genetically engineered wholecell sensor strains; (b) the combination of multiple strains into a single sensor panel to effect multiple response modes; (c) development of a computerized algorithm to analyze the panel responses; (d) laboratory testing and calibration; (e) field testing. In the course of the project, mostly due to the change in the US partner, three modifications were introduced to the original objectives: (a) the scope of the project was expanded to include pharmaceuticals (with a focus on antibiotics) in addition to endocrine disrupting chemicals, (b) the computerized algorithm was not fully developed and (c) the field test was not carried out. Major achievements: (a) construction of innovative bacterial sensor strains for accurate and sensitive detection of agriculturally-relevant pollutants, with a focus on endocrine disrupting compounds (UK and HUJ) and antibiotics (HUJ); (b) optimization of methods for long-term preservation of the reporter bacteria, either by direct deposition on solid surfaces (HUJ) or by the construction of spore-forming Bacillus-based sensors (UK); (c) partial development of a computerized algorithm for the analysis of sensor panel responses. Implications: The sensor panel developed in the course of the project was shown to be applicable for the detection of a broad range of antibiotics and EDCs. Following a suitable development phase, the panel will be ready for testing in an agricultural environment, as an innovative tool for assessing the environmental impacts of EDCs and pharmaceuticals. Furthermore, while the current study relates directly to issues of water quality and soil health, its implications are much broader, with potential uses is risk-based assessment related to the clinical, pharmaceutical, and chemical industries as well as to homeland security.
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