Bibliografie tematiche / In-Situ fluorescence microscopy

Letteratura scientifica selezionata sul tema "In-Situ fluorescence microscopy"

Autore: Grafiati

Pubblicato: 25 maggio 2024

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Articoli di riviste sul tema "In-Situ fluorescence microscopy":

Lu, Fang, Tingting Zhou, Yan Liu, Liying Song, Bin Zhang e Yuyan Li. "Application of Fluorescence In Situ Hybridization Assisted by Fluorescence Microscope in Detection of Her2 Gene in Breast Cancer Patients". Contrast Media & Molecular Imaging 2022 (11 agosto 2022): 1–6. http://dx.doi.org/10.1155/2022/3087681.

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In order to study the important factors for evaluating the prognosis of breast cancer patients, a fluorescence microscopy-assisted fluorescence in situ hybridization technique was proposed. Compared with other detection techniques, fluorescence in situ hybridization (FISH) technology assisted by a fluorescence microscope has gradually gained favor in related fields due to its advantages of high detection specificity, high sensitivity, and strong experimental period. Combined with the basic overview of fluorescence microscopy and FISH technology, the advantages and application points of FISH technology assisted by fluorescence microscopy in the detection of the Her2 gene in breast cancer patients were studied and discussed. The results show that IHC can be used as the primary screening for HER2 gene status detection; IHC (2+) and IHC (3+) have false positives, which are related to chromosome 17 polysomy, so FISH should be done to confirm the diagnosis.

SERWER, PHILIP, SHIRLEY J. HAYES, KAREN LIEMAN e GARY A. GRIESS. "In situ fluorescence microscopy of bacteriophage aggregates". Journal of Microscopy 228, n. 3 (dicembre 2007): 309–21. http://dx.doi.org/10.1111/j.1365-2818.2007.01855.x.

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Liv, Nalan, Daan S. B. van Oosten Slingeland, Jean-Pierre Baudoin, Pieter Kruit, David W. Piston e Jacob P. Hoogenboom. "Electron Microscopy of Living Cells During in Situ Fluorescence Microscopy". ACS Nano 10, n. 1 (8 dicembre 2015): 265–73. http://dx.doi.org/10.1021/acsnano.5b03970.

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Ballard, S. G., e D. C. Ward. "Fluorescence in situ hybridization using digital imaging microscopy." Journal of Histochemistry & Cytochemistry 41, n. 12 (dicembre 1993): 1755–59. http://dx.doi.org/10.1177/41.12.8245423.

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Bouffier, Laurent, e Thomas Doneux. "Coupling electrochemistry with in situ fluorescence (confocal) microscopy". Current Opinion in Electrochemistry 6, n. 1 (dicembre 2017): 31–37. http://dx.doi.org/10.1016/j.coelec.2017.06.015.

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Leger, I., M. Robert-Nicoud e G. Brugal. "Combination of DNA in situ hybridization and immunocytochemical detection of nucleolar proteins: a contribution to the functional mapping of the human genome by fluorescence microscopy." Journal of Histochemistry & Cytochemistry 42, n. 2 (febbraio 1994): 149–54. http://dx.doi.org/10.1177/42.2.8288860.

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The recent application of DNA cloning and non-radioactive in situ hybridization techniques has strengthened the hypothesis of an ordered chromatin structure in interphase nuclei. The arrangement of specific chromosomal regions is not random and is strongly suspected to vary with functional activity. The combination of in situ hybridization and immunocytochemistry, allowing simultaneous detection of nucleic acid sequences and specific antigens in the same nucleus, has already made significant contributions to the study of gene expression, to simultaneous karyotyping and phenotyping of tumor cells, and to in situ analysis of viral infections. This report emphasizes the considerable interest of such combined techniques for functional in situ mapping of the genome at the individual cell level. We propose a method that combines fluorescence immunocytochemical detection of nucleolar proteins and fluorescence in situ hybridization of centromeric and telomeric probes specific for chromosome 1 in two cultured human cell lines. The preparative constraints for a broad application of this procedure are defined so that the cell preparations can be further analyzed by fluorescence microscopic imaging techniques and confocal laser scan microscopy. The two selected sequences of the human chromosome 1 can be localized in the nucleus with respect to nucleolar proteins in a one-step fluorescence microscopic observation.

Reinhardt, Susanne C. M., Luciano A. Masullo, Isabelle Baudrexel, Philipp R. Steen, Rafal Kowalewski, Alexandra S. Eklund, Sebastian Strauss et al. "Ångström-resolution fluorescence microscopy". Nature 617, n. 7962 (24 maggio 2023): 711–16. http://dx.doi.org/10.1038/s41586-023-05925-9.

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AbstractFluorescence microscopy, with its molecular specificity, is one of the major characterization methods used in the life sciences to understand complex biological systems. Super-resolution approaches1–6 can achieve resolution in cells in the range of 15 to 20 nm, but interactions between individual biomolecules occur at length scales below 10 nm and characterization of intramolecular structure requires Ångström resolution. State-of-the-art super-resolution implementations7–14 have demonstrated spatial resolutions down to 5 nm and localization precisions of 1 nm under certain in vitro conditions. However, such resolutions do not directly translate to experiments in cells, and Ångström resolution has not been demonstrated to date. Here we introdue a DNA-barcoding method, resolution enhancement by sequential imaging (RESI), that improves the resolution of fluorescence microscopy down to the Ångström scale using off-the-shelf fluorescence microscopy hardware and reagents. By sequentially imaging sparse target subsets at moderate spatial resolutions of >15 nm, we demonstrate that single-protein resolution can be achieved for biomolecules in whole intact cells. Furthermore, we experimentally resolve the DNA backbone distance of single bases in DNA origami with Ångström resolution. We use our method in a proof-of-principle demonstration to map the molecular arrangement of the immunotherapy target CD20 in situ in untreated and drug-treated cells, which opens possibilities for assessing the molecular mechanisms of targeted immunotherapy. These observations demonstrate that, by enabling intramolecular imaging under ambient conditions in whole intact cells, RESI closes the gap between super-resolution microscopy and structural biology studies and thus delivers information key to understanding complex biological systems.

Collinson, Lucy M. "Smart Microscopy: Automation of CLEM using In situ Fluorescence Detection". Microscopy and Microanalysis 25, S2 (agosto 2019): 1018–19. http://dx.doi.org/10.1017/s1431927619005828.

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Arend, J., A. Wetzel e B. Middendorf. "In-situ-investigation of superplasticizer-particle-interaction by fluorescence microscopy". Materials Today: Proceedings 5, n. 7 (2018): 15292–97. http://dx.doi.org/10.1016/j.matpr.2018.05.008.

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Fetni, Raouf, Patrick Scott, Frédérique Tihy, Claude-Lise Richer e Nicole Lemieux. "Increased resolution of in situ hybridization signal by electron microscopy: A comparison with fluorescence microscopy". Genome 42, n. 5 (1 ottobre 1999): 1001–7. http://dx.doi.org/10.1139/g99-071.

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Cytogenetic studies by in situ hybridization (ISH) have proven to be valuable for gene mapping on banded chromosomes when combined with fluorescence microscopy (FISH). However, even under the best conditions, FISH technology has a resolving power inherent to light of just 0.2 µm. Its utilization is further limited by the diffusion of light coming from the fluorescent signal which covers an area considerably larger than the target DNA sequence. The development of new ISH protocols applied to electron microscopy (EMISH) should increase the resolution for cytogenetic mapping and fine chromosomal structure studies. Despite these advances, few attempts have been made which exploit this increased resolution. Here we present a detailed analysis of ISH signals obtained by fluorescence and electron microscopy methodologies to demonstrate and define the higher sensitivity obtainable by electron microscopy. This comparative study was conducted with probes of different origins: telomeric, classical satellite, alpha satellite, and single-copy DNA sequences, which provide a good reference point for later studies. We were also able to map a 200-bp cDNA probe by EMISH. This study assesses the nature of the resolution and the better definition of the EMISH signal, which confirms the greater resolution of electron microscopy as compared with that achieved with light microscopy. It also indicates that better delineation of two closely linked sequences is achieved at the electron microscopy level.Key words: In situ hybridization, electron microscopy, fluorescence microscopy, localization, repetitive and small single-copy probes.

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Tesi sul tema "In-Situ fluorescence microscopy":

Man, Hiu Mun. "Characterisation of enzymatic catalysis by microscopy and electrochemistry : application to H2/O2 bio-fuel cells". Electronic Thesis or Diss., Aix-Marseille, 2022. http://theses.univ-amu.fr.lama.univ-amu.fr/221207_MAN_82cby815lbx134rmsegm855nh_TH.pdf.

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Les biopiles enzymatiques, qui utilisent des enzymes pour convertir l'énergie chimique en électricité, se présentent comme l'une des ressources énergétiques alternatives et propres les plus prometteuses. Cependant, l'immobilisation fonctionnelle de ces enzymes sur une électrode pour une catalyse efficace suscite encore de nombreux défis. Afin d’accéder à des informations résolues dans l'espace, il est nécessaire de coupler l'électrochimie à d’autres techniques de surface. Dans cette thèse, la microscopie de fluorescence confocale à balayage laser a été couplée à l'électrochimie pour la caractérisation de la catalyse électro-enzymatique. La principale réaction étudiée était la réaction de réduction de l'oxygène catalysée par la bilirubine oxydase de Myrothecium verrucaria. Cette réaction implique une consommation de protons couplée au transfert d'électrons. En utilisant une analyse in situ, les variations locales de pH qui se produisent à proximité de la bioélectrode pendant la catalyse enzymatique sont visualisées grâce à un fluorophore dont l’émission dépend du pH, la fluorescéine. L'activité de l'enzyme a d’abord été sondée par spectroscopie UV-vis et électrochimie. Nous avons ensuite montré que l'intensité de la fluorescence enregistrée est directement proportionnelle au courant catalytique. Les profils d'appauvrissement en protons à l’interface électrochimique dans des électrolytes tamponnés et non tamponnés ont été reconstruits, afin de déterminer l'influence de la force ionique sur l'environnement local des enzymes. Enfin, les enzymes ont été marquées avec des fluorophores, permettant de révéler les hétérogénéités locales de leur distribution interfaciale
Enzyme biofuel cells, which use enzymes to convert chemical energy into electricity, hold promise as one of the most promising alternative and clean energy resources. However, the immobilization of such enzymes on an electrode for efficient catalysis still raises many challenges. In order to access spatially resolved information, it is necessary to couple electrochemistry to other surface techniques. In this thesis, confocal laser scanning fluorescence microscopy was coupled with electrochemistry for the characterization of electro-enzymatic catalysis. The main reaction studied was the oxygen reduction reaction catalyzed by bilirubin oxidase from Myrothecium verrucaria. This reaction involves a consumption of protons coupled with electron transfer. Using in situ analysis, the local pH variations that occur near the bioelectrode during the enzymatic catalysis are visualized thanks to a fluorophore whose emission depends on the pH, fluorescein. The activity of the enzyme was first probed by UV-vis spectroscopy and electrochemistry. We then showed that the intensity of the fluorescence recorded is directly proportional to the catalytic current. Profiles of proton depletion at the electrochemical interface in buffered and unbuffered electrolytes were reconstructed to determine the influence of ionic strength on the local environment of enzymes. Finally, the enzymes were labeled with fluorophores, making it possible to reveal the local heterogeneities of their interfacial distribution

Roussille, Ludovic. "Suivi quantitatif in situ d'interactions biomoléculaire par microscopie optique SEEC". Thesis, Le Mans, 2012. http://www.theses.fr/2012LEMA1030/document.

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La microscopie SEEC (Surface Enhanced Ellipsometric Contrast) est une technique inventée au mans, il y a une dizaine d’années. Elle permet de visualiser des objets de taille nanoscopique entre polariseur et analyseur croisés en utilisant les propriétés non dépolarisantes de surfaces multicouches. Jusqu’au début de la thèse, seules des observations à l’air étaient possibles. Le but de cette thèse a consisté à adapter cette technique à l’observation in-situ d’objets en immersion dans l’eau.Pour cela, il a fallu inventer de nouvelles surfaces propres à ce nouveau milieu. Les calculs ont montrés que des surfaces fines d’or révélaient un bon contraste pour des objets de 1 nm en immersion dans l’eau. Expérimentalement, nous avons montré que pour exploiter au maximum le contraste SEEC, il est nécessaire de modifier l’éclairage. En parallèle de ces travaux expérimentaux, de nouveaux calculs ont montrés que l’utilisation d’épaisseurs encore plus fines permettait de visualiser ces objets avec un bon contraste et sans aucune modification de l’éclairage. Nous avons appelé cette nouvelle technique : la microscopie CONE. Nous avons découvert deux modes de mesure. Après avoir réalisé des fonctionnalisations homogènes et hétérogènes des surfaces d’or. Ces surfaces ont été utilisées en résonance plasmonique de surface (SPR) pour l’étude de fixation de protéine (adsorption et immobilisation) puis d’interaction protéine/protéine. Ces expériences ont ensuite servies de référence pour évaluer les microscopies SEEC et CONE. Par cela, nous avons prouvé que ces microscopies présentent de forts intérêts pour la détection in-situ de protéines avec un faible coût
This thesis was supported by National Agency for Research with the project: ANR PNANO-07 SEEC. The Surface Enhanced Ellipsometric Contrast (SEEC) microscopy has invented in 2000 at Le Mans (France). This technique allows the visualization of nanoscopic object between crossed analyzer and polarizer. It’s possible if some special multilayer surfaces are used. There surfaces must have the particularity to not change the polarization of light during the reflection. Until the beginning of the project the SEEC microscopy was useful only for air observations. The goal of the thesis was to adapt this technique to observe on gold surfaces immerged in water and to compare the performance of the SEEC microscopy with Surface Plasmonique Resonance (SPR) in that configuration. The SPR is a biomolecular interaction study reference technique. SEEC microscopy lateral resolution was evaluate by fluorescence microscopy. Next, we realize two model experiments monitor in parallel by SEEC microscopy and by SPR: BSA immobilization and biotinylated IgG fixation by immobilized streptavidine. To compare measurements efficiently we did a huge preparation work (surface functionalizations and microfluidic) to have exactly same conditions in both techniques.Our results show SEEC microscopy cannot replace SPR for biomolecular interaction studies but it can be used as cheap immunological diagnostic technique. This work gives the path to follow on that direction

Ashok, Mahima. "Analysis of HER2 testing in breast cancer". Diss., Atlanta, Ga. : Georgia Institute of Technology, 2009. http://hdl.handle.net/1853/29711.

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Thesis (Ph.D)--Biomedical Engineering, Georgia Institute of Technology, 2010.
Committee Chair: Griffin, Paul; Committee Member: Butera, Robert; Committee Member: Halpern, Michael; Committee Member: Nichols, Richard; Committee Member: Vidakovic, Brani. Part of the SMARTech Electronic Thesis and Dissertation Collection.

Yara, Ricardo. "Localização in situ e caracterização molecular da bactéria endossimbionte de Pleurotus ostreatus". Universidade de São Paulo, 2006. http://www.teses.usp.br/teses/disponiveis/11/11137/tde-21082006-150238/.

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O fungo Pleurotus ostreatus pertence ao grupo de basidiomicetos que degradam madeira. Este cogumelo cultivado em todo mundo apresenta grande rusticidade e produtividade, e pode ainda ser usado em processos de biorremediação e biopolpação. Devido a seu potencial biotecnológico, torna-se interessante a compreensão da interação deste com outros microrganismos. Neste sentido, recentemente foi observada a presença de bactérias associadas a P. ostreatus em culturas in vitro, que apresentavam grande pleomorfismo. A partir desta observação foram elaborados ensaios que visaram a confirmação da presença de bactérias. Para tanto, foi utilizada a estratégia do Ciclo Completo de Análise do rRNA (full-cycle rRNA analysis) empregada em microrganismos não cultiváveis ou de crescimento fastidioso, além do emprego de técnicas de microbiologia básica, e de estudos de ultraestrutura. Os estudos de microbiologia básica indicaram que se tratava de um microrganismo fastidioso e que se desenvolvia melhor na presença do fungo em sistema de co-cultivo em meios contendo Tween 80 ou Tween 20. Por sua vez, a análise de ultraestrutura demonstrou a presença de estruturas pleomórficas, tanto internamente como externamente à hifa. Em relação ao Ciclo completo de Análise do rRNA este se iniciou pela amplificação e seqüenciamento de parte do rDNA bacteriano, que revelou a proximidade desta bactéria com o Complexo Burkholderia cepacia (CBC). A partir desta seqüência, foi realizado um estudo de bioinformática que indicou sondas específicas para este grupo de bactérias. Completando o Ciclo completo de Análise do rRNA, foram realizados ensaios de hibridização in situ fluorescente (FISH) para a confirmar a relação entre as estruturas bacterianas e a seqüência obtida. Este método comprovou a presença das bactérias no interior das hifas de P. ostreatus. Este trabalho constitui o primeiro relato de bactérias pleomórficas pertencente ao complexo B. cepacia associados a P. ostreatus.
The fungus Pleurotus ostreatus, which belongs to white rot basidiomycete group, is a widely cultivated mushroom; this species has high productivity and rusticity, besides its use in biobleaching and bioremediation processes. This biotechnological potential justifies microbial interaction studies between this fungi and others microorganisms. In P. ostreatus mycelia, it has been observed pleomorphic bacteria growing on agar media. This research describes several assays to confirm bacterial presence in this sample. Therefore, the full-cycle rRNA analysis (described for unculturable or fastidious microorganism), ultrastructure and basic microbiology approaches were employed. Basic microbiology approaches indicated slow growing bacteria, which grown faster near to fungi colonies in solid media amended with Tween 80 or Tween 20 (co-culture system). Ultrastructure studies confirm the presence of intracellular and extracellular pleomorphic bacteria. The full-cycle rRNA analysis started with 16S rDNA amplification and sequencing. This approach demonstrated a relation between these bacteria with Burkholderia cepacia complex. By bioinformatics analysis was determinate which DNA probes can be use to identified this bacterial group. The last step for full-cycle rRNA analysis was applying fluorescent in situ hybridization (FISH). This technique confirmed the relationship between 16S rDNA bacterial sequence and bacterial forms. This is the first time that a pleomorphic bacteria from B. cepacia complex is found associated with P. ostreatus.

Emad, Ahmed Anwar Hasanin. "Development and assessment of strategies for non-invasive prenatal diagnosis using fetal cells in maternal blood". Thèse, Université de Sherbrooke, 2014. http://hdl.handle.net/11143/5855.

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Abstract : Current prenatal diagnosis depends on invasive procedures and is thus offered only to high-risk pregnancies. Development of non-invasive prenatal diagnosis (NIPD) would change the risk-benefit ratio and make it likely that more women would benefit from prenatal testing. Scientists have documented the presence of rare fetal cells in maternal blood and envisioned targeting them with specific markers and their use in NIPD. Considering their extremely low frequency in maternal blood, fetal cells have been difficult to retrieve and use in clinical practice. Therefore, there is a pressing need for systematic sequential studies to evaluate their feasibility in NIPD. Generally, detection of rare cells within a large cell population carries great potentialities for the prospects of cancer management and NIPD. Manual scanning is very cumbersome and time-consuming Therefore; the first part of our project was, dedicated to the optimization of an effective strategy to evaluate retrieval of rare cells. We have developed a way of distributing a controlled number of target cells among hundreds of thousands of other cells on microscope slides. This strategy allows the precise evaluation of the retrieval of rare events and the comparizon of the efficacy of different techniques and enrichment approaches by knowing the definite number and locations of target cells on the slides. Furthermore, it allows the evaluation of hybridization of missed events. We have also developed a robust custom-made detection algorithm for rare cells using the MetaSystems automated platform and have used this strategy in the validation of manual and automatic scanning of 60 slides with a pre-defined number of rare male cells among a pure population of female cells using XY-FISH. Consequently, we tested the developed classifier for the detection of real fetal cells from maternal blood in both normal and aneuploid pregnancies with Down syndrome. We further evaluated the number of fetal cells with different methods of enrichments in the first and second trimesters. The data collected confirmed the early presence of fetal cells in all of the pregnancies tested and their frequencies were higher in cases of aneuploidies. Fetal cells are in a state of dynamic change throughout the pregnancy. Higher numbers of these cells can be obtained by optimizing the harvest time and methods of enrichment. We found that automatic scanning is more sensitive and reliable than manual detection. Furthermore, it alleviates the burden of scanning large numbers of cells and thus is more suitable for clinical application. We also demonstrated the feasibility of using rare cells in NIPD. Five microdissected amniotic fetal cells from 26 cases of normal and aneuploid pregnancies were quite enough to provide accurate NIPD through using whole genome amplification coupled with QF-PCR. Our findings laid the ground for the use of rare fetal cells in maternal blood for NIPD. // Résumé : Le diagnostic prénatal résulte encore aujourd’hui de procédures invasives, qui présentent des risques pour la grossesse. Le développement du diagnostic prénatal non-invasif (DPNI) changerait le rapport risque : bénéfice, rendant le diagnostic prénatal plus intéressant pour les femmes enceintes. Plusieurs chercheurs ont montré la présence de cellules fœtales dans le sang maternel et des travaux ont été entrepris afin de les cibler et de les utiliser éventuellement en DPNI. Toutefois, la faible concentration des cellules fœtales dans le sang maternel réduit les possibilités d’isolement ainsi que celles de leur utilisation en clinique. Un autre aspect technique du DPNI, le balayage manuel, est très laborieux, surtout en terme de temps technique. Il y a donc un besoin certain pour des études approfondies afin d’évaluer et d’améliorer la faisabilité du DPNI. La détection d’évènements rares dans une grande population cellulaire offre un potentiel pour le diagnostic en oncologie mais aussi en diagnostic prénatal. Dans cette thèse, la première étude était dédiée à l’optimisation d’une stratégie pour détecter les cellules rares. Nous avons développé une méthode d’étalement sur lame d’un nombre précis de cellules cibles parmi des centaines de milliers de cellules. Cette stratégie a permis d’évaluer le taux de détection d’évènements rares et de comparer l’efficacité des techniques d’enrichissement en connaissant le nombre exact et la localisation de cellules cibles sur les lames. De plus, il a été possible d’évaluer les problèmes d’hybridation des évènements manqués. Nous avons, par la suite, développé un algorithme robuste pour la détection de cellules rares en utilisant la plateforme de microscopie automatisée MetaSystems et utilisé cette approche dans la validation des balayages manuel et automatique d’un nombre précis de cellules mâles parmi une large population de cellules femelles marquées avec la technique FISH. Nous avons testé ce classificateur avec des échantillons de sang de femmes enceintes de grossesses normales et aneuploïdes et évalué la fréquence de cellules fœtales isolées par différentes méthodes d’enrichissement au cours des premier et second trimestres de grossesse. Les données accumulées ont confirmé la présence de cellules fœtales chez toutes les grossesses et leur fréquence plus élevée dans les grossesses aneuploïdes. Le nombre de cellules fœtales est dynamique tout au long de la grossesse. De plus, un nombre plus élevé de cellules fœtales peut être obtenu en optimisant le moment du prélèvement et les méthodes d’enrichissement. De plus, le balayage automatique s’est avéré plus sensible et constant que le balayage manuel, ce qui permet de balayer un grand nombre de cellules et devient plus approprié pour une application clinique. Nous avons aussi montré la faisabilité d’utiliser des cellules fœtales dans le cadre du DPNI. Cinq cellules amniotiques microdisséquées, provenant de grossesses normales et aneuploïdes, ont suffi pour poser un diagnostic prénatal par une combinaison de l’amplification du génome complet et de la technique QF-PCR (réaction quantitative en fluorescence d’amplification entraînée par une polymérase) permettant la détection d’anomalies chromosomiques. Nos résultats ouvrent la voie à l’utilisation de cellules fœtales dans le sang maternel pour le DPNI.

Galanti, Agostino. "Multi-photochromic architectures : from structure to function". Thesis, Strasbourg, 2018. http://www.theses.fr/2018STRAF046/document.

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L’objectif de cette thèse a été axé sur le développement des systèmes capable de répondre à des stimuli externes, basés sur des unités photochromiques. Le but d’une telle quête est d’augmenter la complexité des dispositifs et des machines moléculaires synthétiques. Avec l’objectif de développer des dispositifs et des machines artificiels plus complexes, nous avons réalisé de systèmes comprenant de multiples interrupteurs moléculaires. En vue de la réalisation de cette thèse, des nouveaux systèmes multi-photochromiques, où hybrides photochrome/nanomatériaux contenant des fragments azobenzène, diaryléthène ou spiropyrane ont été réalisés et étudiés. D’abord, on s’est focalisés sur des systèmes multi-azobenzènes capables de subir de grands réarrangements géométriques lors de la photoisomérisation, ils pourraient être utilisés à l'avenir comme éléments constitutifs des matériaux host-guest ou metal-organic frameworks contrôlables par des stimuli lumineux. Dans un second exemple, des commutateurs photochromiques de type dithiényléthène ont été utilisés pour déclencher l'émission d'une porphyrine. Cette dyade à montré une modulation réversible de son émission, affichant un contraste particulièrement élevé. Comme dernier exemple, un dérivé de spiropyrane a été combiné avec des nanoparticules d’or anisotropes. En induisant l'isomérisation de l’interrupteur moléculaire dans les dispersions colloïdales des nanorods d’or en liquide, nous avons visualisé une grande variation du spectre d'extinction des colloïdes, dépendante de la longueur d’onde du mode LSPR et du recouvrement spectrale avec le photoswitch
The aim of this thesis has been to develop systems capable of responding to external stimuli, based on photochromic units. The goal of such a quest is to increase the complexity of devices and synthetic molecular machines. With the goal of developing more complex artificial devices and machines, we have realised systems containing multiple molecular switches. For the realisation of this thesis, new multi-photochromic systems, or photochromes/nanomaterials hybrids containing azobenzene, diarylethene or spiropyran moieties have been realised and studied. Firstly, we focused on multi-azobenzene systems capable of undergoing large geometric rearrangements during photoisomerisation, as they may be used in the future as constituent elements of host-guest or metal-organic frameworks controllable by luminous stimuli. In a second example, dithienylethene-type photochromic switches have been used to trigger the emission of a porphyrin. This dyad exhibited a reversible modulation of its emission, displaying a particularly highly contrasted response. As a final example, a spiropyran derivative has been combined with anisotropic gold nanoparticles. By inducing the isomerisation of the molecular switch in the AuNR colloidal liquid dispersions, we visualised a large variation of the colloid extinction spectrum, dependent on the LSPR mode wavelength and the spectral overlap with the photoswitch

Mongelard, Fabien. "Apport des approches in situ pour l'analyse du phénomène d'inactivation du chromosome X chez les mammifères". Université Joseph Fourier (Grenoble ; 1971-2015), 1998. http://www.theses.fr/1998GRE10261.

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L'inactivation du chromosome x chez les mammiferes femelles est la mise sous silence transcriptionel d'un des deux chromosomes x. L'organisation de la chromatine du chromosome inactif (xi) est differente de celle de son homologue actif (xa) ; la nature de cette difference est inconnue. Une region contenant le gene xist (x inactive specific transcript) est indispensable au phenomene. Le travail presente ici vise a apporter de nouvelles informations a la fois sur l'organisation de la chromatine des chromosomes actifs et inactifs, et sur la fonction du gene xist. L'analyse de la structure chromatinienne des chromosome x a ete realisee en combinant l'hybridation in situ fluorescente (fish) a la microscopie confocale a balayage laser. Nous avons developpe des outils et des methodes pour evaluer la qualite des protocoles de fish, et pour ameliorer et analyser les images microscopiques numeriques. Des precautions particulieres doivent etre prises pour la quantification des resultats de fish. Dans des noyaux de fibroblastes humains feminins, les deux chromosomes x ont un volume identique mais different par la texture du marquage par fish. Des differences se retrouvent aussi au niveau du centromere. La detection in situ du transcrit xist permet la definition d'une region minimale comprenant xist, capable d'entrainer l'inactivation lors d'experiences de transgenese. L'integration en copie unique d'un transgene contenant xist, contrairement a une integration multiple, est incapable de declencher l'inactivation dans des cellules souches embryonnaires (es) differenciees murines, suggerant que des sequences essentielles a une fonction autonome de la region sont absentes, et indiquant l'existence de facettes insoupconnees du phenomene d'inactivation. Des cellules es de souris transgeniques porteuses de la region contenant le xist humain ont ete etudiees. Une courbure du chromosome cible, est induite au niveau du site d'integration ; la signification de cette courbure est incomprise.

Chen, X. "TAGGING BIOCONTROL STREPTOMYCES TO STUDY LETTUCE COLONIZATION". Doctoral thesis, Università degli Studi di Milano, 2015. http://hdl.handle.net/2434/345187.

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The ability of the biological control agents (BCAs) to colonize plant tissues is an important feature involved in microbe-assisted plant protection. Plant-microbe interaction research increased especially in the last decade thanks to technological revolution. Molecular methods and the development of advanced microscopic techniques allow researchers to explore gene expression and localization of beneficial microorganisms within plants. The green fluorescent protein (GFP) and its modified version, enhanced GFP (EGFP), more adapt for expression in mammalian cells and GC-rich actinomycetes like Streptomyces, have been widely used as markers to study gene expression, as well as plant-microbe interactions. Aside fluorescent protein approaches, fluorescence in situ hybridization (FISH) is another frequently used technique to visualize microbial colonization patterns and community composition by application of specific fluorescent probes. Firstly, we transformed five Streptomyces strains, which showed strong inhibition activity against Sclerotinia sclerotiorum, with the EGFP construct by the conjugation method. The conjugation efficiencies varied between the strains, but were comparable to the reference strain. The fitness of transformed strains was similar to wild-type; the transformants maintained similar sporulation, mycelium growth rate, and the ability to produce important secondary metabolites and lytic enzymes. Secondly, two transformed strains, Streptomyces cyaneus ZEA17I, and Streptomyces sp. SW06W, were used to study lettuce colonization dynamics by seed coating method. Their spatio-temporal dynamics were determined in sterile substrate. The strains were consistently recovered from lettuce rhizosphere and inner root tissues up to six weeks. Finally, the colonization pattern of lettuce by Streptomyces cyaneus ZEA17I was examined by both EGFP and FISH approaches combined with confocal laser scanning microscopy (CLSM). For FISH-CLSM analysis, universal bacteria and Streptomyces genus specific probes were used to label S. cyaneus ZEA17I. The consistent presence of the labeled strain at the lettuce root one week after sowing showed that Streptomyces spores could rapidly germinate and produce filamentous mycelium on lettuce. S. cyaneus ZEA17I was detected also on two-week-old roots, indicating the long-term survival ability of this strain in lettuce rhizosphere. Altogether, the antagonistic activity, rhizosphere and root competence showed by the Streptomyces conferred their potential to act as BCA. Further studies on the complex host-pathogen-antagonist interactions will provide additional knowledge to understand the modes and mechanisms of Streptomyces-mediated plant protection.

PERRIN, CHRISTELE. "Methodologie pour l'analyse quantitative en imagerie microscopique conventionnelle et a fluorescence. Application a l'etude de la proliferation et de l'expression du recepteur a l'egf dans des cellules tumorales mammaires". Université Joseph Fourier (Grenoble), 1996. http://www.theses.fr/1996GRE10198.

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L'utilisation accrue des methodes d'imagerie microscopique dans le domaine de la cancerologie, et les developpements recents des marqueurs specifiques utilises dans ce contexte ont fait naitre un besoin urgent en ce qui concerne le developpement de procedures de multi-marquages enzymatiques ou fluorescents, et leur quantification cellule a cellule. C'est dans ce contexte de mise au point methodologique qu'ont ete realises les travaux presentes dans cette these. Ils incluent les divers aspects de la standardisation de methodes de multi-marquages, l'adaptation des methodes d'imagerie pour la quantification du marquage in situ du recepteur a l'egf, et la normalisation des mesures. Les techniques developpees pour la quantification par cytometrie en image sont detaillees, faisant apparaitre les limites et les problemes eventuels rencontres, ainsi que la facon de les contourner ou de les resoudre. Ces developpements methodologiques ont ete appliques a l'etude de l'analyse quantitative in situ des recepteurs a l'egf dans les tissus animaux et humains (tumeurs mammaires), en relation avec la cinetique de proliferation de chacune des cellules (marquages ki67, brdu, agnor). L'etude de marquages simultanes des proteines ki67-agnors-egfr associes au marquage de l'adn montre qu'il devrait etre possible de preciser pour chaque cellule en cycle, dans un tissu normal ou tumoral, la relation entre la vitesse du cycle et l'expression des recepteurs a l'egf

KOMATSU, LUIZ G. H. "Estudo comparativo de nanocompósitos de polipropileno modificado sob condições de envelhecimento ambiental e acelerado". reponame:Repositório Institucional do IPEN, 2016. http://repositorio.ipen.br:8080/xmlui/handle/123456789/26380.

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Submitted by Claudinei Pracidelli (cpracide@ipen.br) on 2016-06-22T12:34:18Z No. of bitstreams: 0
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Dissertação (Mestrado em Tecnologia Nuclear)
IPEN/D
Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

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Libri sul tema "In-Situ fluorescence microscopy":

Bridger, Joanna M., e Emanuela V. Volpi. Fluorescence in situ hybridization (FISH): Protocols and applications. New York, NY: Humana Press, 2010.

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Liehr, Thomas. Fluorescence In Situ Hybridization (FISH) — Application Guide. Berlin, Heidelberg: Springer Berlin Heidelberg, 2009.

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1923-, Sharma Arun Kumar, e Sharma Archana 1932-, a cura di. Chromosome painting: Principles, strategies, and scope. Dordrecht: Kluwer Academic Publishers, 2001.

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Fried, Alexander. Studies of phase transitions in supported lipid bilayers by simultaneous in situ fluorescence spectroscopy and atomic force microscopy. Ottawa: National Library of Canada, 2002.

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A, Sharif N., a cura di. Molecular imaging in neuroscience: A practical approach. Oxford [England]: IRL Press at Oxford University Press, 1993.

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Liehr, Thomas. Fluorescence in Situ Hybridization: Application Guide. Springer, 2016.

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Liehr, Thomas. Fluorescence In Situ Hybridization - Application Guide. Springer, 2010.

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Liehr, Thomas. Fluorescence in Situ Hybridization: Application Guide. Springer Berlin / Heidelberg, 2018.

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(Editor), Arun Kumar Sharma, e Archana Sharma (Editor), a cura di. Chromosome Painting: Principles, Strategies and Scope. Springer, 2001.

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Sharma, Arun Kumar, e Archana Sharma. Chromosome Painting: Principles, Strategies and Scope. Springer London, Limited, 2012.

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Capitoli di libri sul tema "In-Situ fluorescence microscopy":

Michalová, Kyra, Zuzana Zemanová, Jana Březinová e Věra Michalová. "Fluorescence in Situ Hybridization (FISH) in Cytogenetics of Leukemia". In Fluorescence Microscopy and Fluorescent Probes, 185–89. Boston, MA: Springer US, 1996. http://dx.doi.org/10.1007/978-1-4899-1866-6_27.

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Rinke, Bernd, Joachim Bradl, Bernhard Schneider, Markus Durm, Michael Hausmann, Horst Ludwig e Christoph Cremer. "“In Situ” Estimates of the Spatial Resolution for “Practical” Fluorescence Microscopy of Cell Nuclei". In Fluorescence Microscopy and Fluorescent Probes, 169–73. Boston, MA: Springer US, 1996. http://dx.doi.org/10.1007/978-1-4899-1866-6_24.

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Iourov, Ivan Y. "Microscopy and Imaging Systems". In Fluorescence In Situ Hybridization (FISH) — Application Guide, 75–84. Berlin, Heidelberg: Springer Berlin Heidelberg, 2009. http://dx.doi.org/10.1007/978-3-540-70581-9_7.

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Markaki, Yolanda, Daniel Smeets, Marion Cremer e Lothar Schermelleh. "Fluorescence In Situ Hybridization Applications for Super-Resolution 3D Structured Illumination Microscopy". In Nanoimaging, 43–64. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-62703-137-0_4.

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Edwards, Matthew S. "Using in situ substratum sterilization and fluorescence microscopy in studies of microscopic stages of marine macroalgae". In Sixteenth International Seaweed Symposium, 253–59. Dordrecht: Springer Netherlands, 1999. http://dx.doi.org/10.1007/978-94-011-4449-0_29.

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Smolka, John A., e Samantha C. Lewis. "In Situ Analysis of Mitochondrial DNA Synthesis Using Metabolic Labeling Coupled to Fluorescence Microscopy". In Methods in Molecular Biology, 99–106. New York, NY: Springer US, 2023. http://dx.doi.org/10.1007/978-1-0716-2922-2_8.

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Casanova-Moreno, Jannu, Zhinan Landis Yu, Jonathan Massey-Allard, Brian Ditchburn, Jeff F. Young e Dan Bizzotto. "In Situ Spectroelectrochemical Fluorescence Microscopy for Visualizing Interfacial Structure and Dynamics in Self-assembled Monolayers". In Luminescence in Electrochemistry, 21–77. Cham: Springer International Publishing, 2017. http://dx.doi.org/10.1007/978-3-319-49137-0_2.

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Grohme, Markus A., Olga Frank e Jochen C. Rink. "Preparing Planarian Cells for High-Content Fluorescence Microscopy Using RNA in Situ Hybridization and Immunocytochemistry". In Methods in Molecular Biology, 121–55. New York, NY: Springer US, 2023. http://dx.doi.org/10.1007/978-1-0716-3275-8_8.

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Schmidt, Hannes, e Thilo Eickhorst. "Gold-FISH: In Situ Hybridization of Microbial Cells for Combined Fluorescence and Scanning Electron Microscopy". In Springer Protocols Handbooks, 545–57. Berlin, Heidelberg: Springer Berlin Heidelberg, 2016. http://dx.doi.org/10.1007/978-3-662-52959-1_53.

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Karygianni, Lamprini, Elmar Hellwig e Ali Al-Ahmad. "Multiplex Fluorescence In Situ Hybridization (M-FISH) and Confocal Laser Scanning Microscopy (CLSM) to Analyze Multispecies Oral Biofilms". In Methods in Molecular Biology, 65–72. New York, NY: Springer New York, 2014. http://dx.doi.org/10.1007/978-1-4939-0467-9_5.

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Atti di convegni sul tema "In-Situ fluorescence microscopy":

Samuel, B. A., e M. A. Haque. "In-Situ Nanoscale Single Fiber Fragmentation Using Fluorescence Microscopy". In ASME 2007 International Mechanical Engineering Congress and Exposition. ASMEDC, 2007. http://dx.doi.org/10.1115/imece2007-43367.

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We use fluorescence microscopy to perform real time single fiber fragmentation experiments on individual PolyFurfuryl Alcohol (PFA) nanofibers embedded within a Poly DiMethyl Siloxane (PDMS) matrix. By using fluorescent nanofibers in an optically transparent matrix, fragmentation of the nanoscale fibers (even less than 400 nm) can also be easily visualized using fluorescence emission from the nanowires. When the composite specimen was loaded to saturation strain the fragment lengths distribution was observed to follow a two parameter Weibull frequency distribution. In addition, we also present a digital image correlation based technique to obtain localized strain (and hence stress) data based on the use of fluorescent nanoscale spatial markers.

Son, Jeonghwan, Biagio Mandracchia e Shu Jia. "Miniaturized optical fluorescence microscopy system for parallel in situ imaging". In Frontiers in Optics. Washington, D.C.: OSA, 2020. http://dx.doi.org/10.1364/fio.2020.fw5e.2.

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Kubarev, Alexey. "Development of the in-situ fluorescence microscopy approach to reveal the mechanism of interfacial polymerization of polyamide membranes". In European Microscopy Congress 2020. Royal Microscopical Society, 2021. http://dx.doi.org/10.22443/rms.emc2020.606.

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Bai, Yeran, Zhongyue Guo, ‪Fátima Pereira, Michael Wagner e Ji-Xin Cheng. "Microbial identification and metabolic analysis by mid-infrared photothermal imaging fluorescence in-situ hybridization". In Advanced Chemical Microscopy for Life Science and Translational Medicine 2022, a cura di Garth J. Simpson, Ji-Xin Cheng e Wei Min. SPIE, 2022. http://dx.doi.org/10.1117/12.2611781.

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Qu, Min, e Yanming Zhang. "The study on improving fluorescence microscopy image effects of genomic in situ hybridization". In 2010 3rd International Conference on Biomedical Engineering and Informatics (BMEI). IEEE, 2010. http://dx.doi.org/10.1109/bmei.2010.5640551.

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Klimas, Aleksandra, Brendan R. Gallagher, Emma DiBernardo, Zhangyu Cheng e Yongxin Zhao. "MAGNIFY: molecule anchorable gel-enabled nanoscale in-situ fluorescence microscopy for nanoscale imaging of biomolecules". In Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXX, a cura di Thomas G. Brown, Tony Wilson e Laura Waller. SPIE, 2023. http://dx.doi.org/10.1117/12.2647983.

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Bottiroli, Giovanni F., Piera Balzarini, Anna C. Croce, Donata Locatelli, Simona Vaccino e Carlo Pelliciari. "Fluorescence resonance energy transfer (FRET) microscopy: a tool for in situ study of cellular structures". In International Symposium on Biomedical Optics Europe '94, a cura di Hans-Jochen Foth, Aaron Lewis, Halina Podbielska, Michel Robert-Nicoud, Herbert Schneckenburger e Anthony J. Wilson. SPIE, 1995. http://dx.doi.org/10.1117/12.200887.

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Upadhyay, Jagannath, Daniel S. Park, Karsten E. Thompson e Dimitris E. Nikitopoulos. "3D Measurements of Nano-Particle Transport in Complex 2.5D Micro-Models". In ASME 2015 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2015. http://dx.doi.org/10.1115/imece2015-50635.

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A confocal Micro-Particle Image Velocimetry (C-μPIV) technique along with associated post image processing algorithms is established to quantify three dimensional distributions of nano-particle velocity and concentration at the micro-scale (pore-scale) in 2.5D porous media designed from a Boise rock sample. In addition, an in-situ, non-destructive method for measuring the geometry of the micro-model, including its depth, is described and demonstrated. The particle experiments use 900 nm fluorescence labeled polystyrene particles at a flow rate of 10 nLmin−1 and confocal laser scanning microscopy (CLSM), while in-situ geometry measurements use regular microscope along with Rhodamine dye and a depth-to-fluorescence-intensity calibration. Image post-processing techniques include elimination of background noise and signal from adsorbed nano-particle on the inner surfaces of the micro-model. In addition, a minimization of depth of focus technique demonstrates a capability of optically thin slice allowing us to measure depth wise velocity in 2.5D micro-model. The mean planar components of the particle velocity of the steady-state flow and particle concentration distributions were measured in three dimensions. Particle velocities range from 0.01 to 122 μm s−1 and concentrations from 2.18 × 103 to 1.79 × 104 particles mm−2. Depth-wise results show that mean velocity closer to the top wall is comparatively higher than bottom walls, because of higher planar porosity and smooth pathway for the nano-particles closer to the top wall. The three dimensional micro-model geometry reconstructed from the fluorescence data can be used to conduct numerical simulations of the flow in the as-tested micro-model for future comparisons to experimental results after incorporating particle transport and particle-wall interaction models.

Kung, C.-Y., M. D. Barnes, N. Lermer, W. B. Whitten e J. M. Ramsey. "Confinement, Detection, and Manipulation of Individual Molecules in Attoliter Volumes". In Laser Applications to Chemical and Environmental Analysis. Washington, D.C.: Optica Publishing Group, 1998. http://dx.doi.org/10.1364/lacea.1998.lma.4.

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We report observation of fluorescence from individual rhodamine 6G molecules in streams of charged 1-μm diameter water droplets. With this approach, probe volumes comparable to diffraction-limited fluorescence microscopy1 techniques (≤ 500 attoliters) are achieved, resulting in similarly high contrast between single molecule fluorescence signals and nonfluorescent background. However, since the fluorescent molecules are confined to electrically charged droplets, in situ electrodynamic manipulation can be accomplished in a straightforward manner, allowing experimental control over both the delivery of molecules of interest to the observation region and the laser-molecule interaction time.

McMahon, Nathan, Allison Solanki, Jocelyn Jones, Sunjong Kwon, Young-Hwan Chang, Koei Chin, Michel Nederlof, Joe Gray e Summer L. Gibbs. "Fluorescent Imaging for In Situ Measurement of Drug Target Engagement and Cell Signaling Pathways". In Microscopy Histopathology and Analytics. Washington, D.C.: OSA, 2020. http://dx.doi.org/10.1364/microscopy.2020.mw4a.5.

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