Base de dados : MEDLINE
Pesquisa : E05.588.465 [Categoria DeCS]
Referências encontradas : 9427 [refinar]
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[PMID]:29335514
[Au] Autor:Kaiser M; Jug F; Julou T; Deshpande S; Pfohl T; Silander OK; Myers G; van Nimwegen E
[Ad] Endereço:Biozentrum, University of Basel and Swiss Institute of Bioinformatics, Klingelbergstrasse 50/70, 4056, Basel, Switzerland.
[Ti] Título:Monitoring single-cell gene regulation under dynamically controllable conditions with integrated microfluidics and software.
[So] Source:Nat Commun;9(1):212, 2018 01 15.
[Is] ISSN:2041-1723
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Much is still not understood about how gene regulatory interactions control cell fate decisions in single cells, in part due to the difficulty of directly observing gene regulatory processes in vivo. We introduce here a novel integrated setup consisting of a microfluidic chip and accompanying analysis software that enable long-term quantitative tracking of growth and gene expression in single cells. The dual-input Mother Machine (DIMM) chip enables controlled and continuous variation of external conditions, allowing direct observation of gene regulatory responses to changing conditions in single cells. The Mother Machine Analyzer (MoMA) software achieves unprecedented accuracy in segmenting and tracking cells, and streamlines high-throughput curation with a novel leveraged editing procedure. We demonstrate the power of the method by uncovering several novel features of an iconic gene regulatory program: the induction of Escherichia coli's lac operon in response to a switch from glucose to lactose.
[Mh] Termos MeSH primário: Regulação Bacteriana da Expressão Gênica
Técnicas Analíticas Microfluídicas/métodos
Análise de Célula Única/métodos
Software
[Mh] Termos MeSH secundário: Algoritmos
Rastreamento de Células/instrumentação
Rastreamento de Células/métodos
Escherichia coli/citologia
Escherichia coli/efeitos dos fármacos
Escherichia coli/genética
Glucose/farmacologia
Óperon Lac/genética
Lactose/farmacologia
Análise de Célula Única/instrumentação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
IY9XDZ35W2 (Glucose); J2B2A4N98G (Lactose)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180305
[Lr] Data última revisão:
180305
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180117
[St] Status:MEDLINE
[do] DOI:10.1038/s41467-017-02505-0


  2 / 9427 MEDLINE  
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[PMID]:28456344
[Au] Autor:Dincer C; Bruch R; Kling A; Dittrich PS; Urban GA
[Ad] Endereço:University of Freiburg, Department of Microsystems Engineering (IMTEK), Laboratory for Sensors, Georges-Koehler-Allee 103, 79110 Freiburg, Germany; University of Freiburg, Freiburg Materials Research Center (FMF), Stefan-Meier-Straße 21, 79104 Freiburg, Germany. Electronic address: dincer@imtek.de.
[Ti] Título:Multiplexed Point-of-Care Testing - xPOCT.
[So] Source:Trends Biotechnol;35(8):728-742, 2017 08.
[Is] ISSN:1879-3096
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Multiplexed point-of-care testing (xPOCT), which is simultaneous on-site detection of different analytes from a single specimen, has recently gained increasing importance for clinical diagnostics, with emerging applications in resource-limited settings (such as in the developing world, in doctors' offices, or directly at home). Nevertheless, only single-analyte approaches are typically considered as the major paradigm in many reviews of point-of-care testing. Here, we comprehensively review the present diagnostic systems and techniques for xPOCT applications. Different multiplexing technologies (e.g., bead- or array-based systems) are considered along with their detection methods (e.g., electrochemical or optical). We also address the unmet needs and challenges of xPOCT. Finally, we critically summarize the in-field applicability and the future perspectives of the presented approaches.
[Mh] Termos MeSH primário: Dispositivos Lab-On-A-Chip/tendências
Técnicas Analíticas Microfluídicas
Sistemas Automatizados de Assistência Junto ao Leito/tendências
[Mh] Termos MeSH secundário: Animais
Seres Humanos
Técnicas Analíticas Microfluídicas/instrumentação
Técnicas Analíticas Microfluídicas/métodos
Técnicas Analíticas Microfluídicas/tendências
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW; RESEARCH SUPPORT, NON-U.S. GOV'T
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180306
[Lr] Data última revisão:
180306
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170501
[St] Status:MEDLINE


  3 / 9427 MEDLINE  
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[PMID]:28747398
[Au] Autor:Leivo J; Virjula S; Vanhatupa S; Kartasalo K; Kreutzer J; Miettinen S; Kallio P
[Ad] Endereço:Micro- and Nanosystems Research Group, BioMediTech Institute and Faculty of Biomedical Sciences and Engineering, Tampere University of Technology, Tampere, Finland.
[Ti] Título:A durable and biocompatible ascorbic acid-based covalent coating method of polydimethylsiloxane for dynamic cell culture.
[So] Source:J R Soc Interface;14(132), 2017 Jul.
[Is] ISSN:1742-5662
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Polydimethylsiloxane (PDMS) is widely used in dynamic biological microfluidic applications. As a highly hydrophobic material, native PDMS does not support cell attachment and culture, especially in dynamic conditions. Previous covalent coating methods use glutaraldehyde (GA) which, however, is cytotoxic. This paper introduces a novel and simple method for binding collagen type I covalently on PDMS using ascorbic acid (AA) as a cross-linker instead of GA. We compare the novel method against physisorption and GA cross-linker-based methods. The coatings are characterized by immunostaining, contact angle measurement, atomic force microscopy and infrared spectroscopy, and evaluated in static and stretched human adipose stem cell (hASC) cultures up to 13 days. We found that AA can replace GA as a cross-linker in the covalent coating method and that the coating is durable after sonication and after 6 days of stretching. Furthermore, we show that hASCs attach and proliferate better on AA cross-linked samples compared with physisorbed or GA-based methods. Thus, in this paper, we provide a new PDMS coating method for studying cells, such as hASCs, in static and dynamic conditions. The proposed method is an important step in the development of PDMS-based devices in cell and tissue engineering applications.
[Mh] Termos MeSH primário: Ácido Ascórbico/química
Materiais Revestidos Biocompatíveis/química
Dimetilpolisiloxanos/química
Células Mesenquimais Estromais/fisiologia
Técnicas Analíticas Microfluídicas/instrumentação
[Mh] Termos MeSH secundário: Adesão Celular
Técnicas de Cultura de Células
Proliferação Celular
Sobrevivência Celular
Seres Humanos
Propriedades de Superfície
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Coated Materials, Biocompatible); 0 (Dimethylpolysiloxanes); 63148-62-9 (baysilon); PQ6CK8PD0R (Ascorbic Acid)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180228
[Lr] Data última revisão:
180228
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170728
[St] Status:MEDLINE


  4 / 9427 MEDLINE  
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[PMID]:29220646
[Au] Autor:Hu P; Fabyanic E; Kwon DY; Tang S; Zhou Z; Wu H
[Ad] Endereço:Department of Genetics, University of Pennsylvania, Philadelphia PA 19104, USA; Epigenetics Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia PA 19104, USA.
[Ti] Título:Dissecting Cell-Type Composition and Activity-Dependent Transcriptional State in Mammalian Brains by Massively Parallel Single-Nucleus RNA-Seq.
[So] Source:Mol Cell;68(5):1006-1015.e7, 2017 Dec 07.
[Is] ISSN:1097-4164
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Massively parallel single-cell RNA sequencing can precisely resolve cellular diversity in a high-throughput manner at low cost, but unbiased isolation of intact single cells from complex tissues such as adult mammalian brains is challenging. Here, we integrate sucrose-gradient-assisted purification of nuclei with droplet microfluidics to develop a highly scalable single-nucleus RNA-seq approach (sNucDrop-seq), which is free of enzymatic dissociation and nucleus sorting. By profiling ∼18,000 nuclei isolated from cortical tissues of adult mice, we demonstrate that sNucDrop-seq not only accurately reveals neuronal and non-neuronal subtype composition with high sensitivity but also enables in-depth analysis of transient transcriptional states driven by neuronal activity, at single-cell resolution, in vivo.
[Mh] Termos MeSH primário: Núcleo Celular/metabolismo
Córtex Cerebral/metabolismo
Sequenciamento de Nucleotídeos em Larga Escala
Neurônios/metabolismo
RNA/genética
Convulsões/genética
Análise de Sequência de RNA/métodos
Análise de Célula Única/métodos
Transcrição Genética
[Mh] Termos MeSH secundário: Animais
Núcleo Celular/patologia
Centrifugação com Gradiente de Concentração
Córtex Cerebral/patologia
Córtex Cerebral/fisiopatologia
Modelos Animais de Doenças
Células-Tronco Embrionárias Humanas/metabolismo
Seres Humanos
Cinética
Masculino
Camundongos
Camundongos Endogâmicos C57BL
Técnicas Analíticas Microfluídicas
Células NIH 3T3
Inibição Neural
Neurônios/patologia
Pentilenotetrazol
RNA/metabolismo
Convulsões/metabolismo
Convulsões/patologia
Convulsões/fisiopatologia
Transmissão Sináptica
Transfecção
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
63231-63-0 (RNA); WM5Z385K7T (Pentylenetetrazole)
[Em] Mês de entrada:1712
[Cu] Atualização por classe:180228
[Lr] Data última revisão:
180228
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171209
[St] Status:MEDLINE


  5 / 9427 MEDLINE  
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[PMID]:28459389
[Au] Autor:Sallmon H; Hatch A; Murthy SK; Plouffe BD; Hansmann G
[Ad] Endereço:1 Charité University Medical Center Berlin, Germany.
[Ti] Título:Circulating Endothelial Cell Quantification by Microfluidics Chip in Pulmonary Arterial Hypertension.
[So] Source:Am J Respir Cell Mol Biol;56(5):680-682, 2017 05.
[Is] ISSN:1535-4989
[Cp] País de publicação:United States
[La] Idioma:eng
[Mh] Termos MeSH primário: Células Endoteliais/metabolismo
Hipertensão Pulmonar/metabolismo
Dispositivos Lab-On-A-Chip
Técnicas Analíticas Microfluídicas/instrumentação
Técnicas Analíticas Microfluídicas/métodos
[Mh] Termos MeSH secundário: Animais
Células Endoteliais/patologia
Seres Humanos
Hipertensão Pulmonar/patologia
[Pt] Tipo de publicação:LETTER; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180221
[Lr] Data última revisão:
180221
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170502
[St] Status:MEDLINE
[do] DOI:10.1165/rcmb.2017-0026LE


  6 / 9427 MEDLINE  
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[PMID]:28456636
[Au] Autor:Ramjee MK; Patel S
[Ad] Endereço:Cyclofluidic Limited, BioPark, Broadwater Road, Welwyn Garden City AL7 3AX, United Kingdom. Electronic address: manoj.ramjee@cyclofluidic.co.uk.
[Ti] Título:Continuous-flow injection microfluidic thrombin assays: The effect of binding kinetics on observed enzyme inhibition.
[So] Source:Anal Biochem;528:38-46, 2017 07 01.
[Is] ISSN:1096-0309
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:A microfluidic assay for monitoring the inhibition of thrombin peptidase activity was developed. The system, which utilised soluble reagents in continuous-flow injection mode, was configured so as to allow inhibitor titrations via gradient formation. This microfluidic continuous-flow injection titration assay (CFITA) enabled the potency of a set of small-molecule serine peptidase inhibitors (SPIs) to be evaluated. The results, compared to standard microtiter plate (MTP) data, indicated that a microfluidic CFITA provided an efficient and effective method for evaluating compound potency. Crucially, whereas for fast-acting compounds the rank order of potency between the CFITA and MTP methods was preserved, for slow-acting compounds the observed CFITA potencies were significantly lower. These results, in conjunction with data from computer simulations, clearly demonstrated that continuous-flow assays, and perhaps microfluidic assays in general, must take into account binding kinetics when used to assess reaction criteria.
[Mh] Termos MeSH primário: Antitrombinas/metabolismo
Antitrombinas/farmacologia
Técnicas Analíticas Microfluídicas
Trombina/antagonistas & inibidores
[Mh] Termos MeSH secundário: Substituição de Aminoácidos
Ensaios Enzimáticos
Fluorescência
Cinética
Trombina/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antithrombins); EC 3.4.21.5 (Thrombin)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:180207
[Lr] Data última revisão:
180207
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170501
[St] Status:MEDLINE


  7 / 9427 MEDLINE  
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[PMID]:28450257
[Au] Autor:Carreras MP; Wang S
[Ad] Endereço:Department of Biomedical Engineering, City University of New York - City College, New York, NY 10031, USA.
[Ti] Título:A multifunctional microfluidic platform for generation, trapping and release of droplets in a double laminar flow.
[So] Source:J Biotechnol;251:106-111, 2017 Jun 10.
[Is] ISSN:1873-4863
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Droplet microfluidics, involving micrometer-sized emulsion of droplets is a growing subfield of microfluidics which attracts broad interest due to its application on biological assays. Droplet-based systems have been used as microreactors as well as to encapsulate many biological entities for biomedical and biotechnological applications. Here, a novel microfluidic device is presented for the generation, trapping and release of aqueous including hydrogel droplets in a double laminar oil flow. This platform enables the storage and release of picoliter-sized droplets in two different carrier oils by using hydrodynamic forces without the need of electrical forces or optical actuators. Furthermore, this design allows droplets to be selectively and simultaneously exposed to two different conditions and collected on demand. Successful encapsulation of hepatoma H35 cells was performed on-chip. Viability of cell-laden droplets was performed off-chip to assess the potential applications in 3D encapsulation cell culture and drug discovery assays.
[Mh] Termos MeSH primário: Dispositivos Lab-On-A-Chip
[Mh] Termos MeSH secundário: Alginatos
Animais
Linhagem Celular Tumoral
Sobrevivência Celular
Ácido Glucurônico
Ácidos Hexurônicos
Hidrodinâmica
Hidrogéis
Técnicas Analíticas Microfluídicas
Óleo Mineral
Peptídeos
Ratos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Alginates); 0 (Hexuronic Acids); 0 (Hydrogels); 0 (Peptides); 0 (RADA16-I); 8020-83-5 (Mineral Oil); 8A5D83Q4RW (Glucuronic Acid); 8C3Z4148WZ (alginic acid)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180205
[Lr] Data última revisão:
180205
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170429
[St] Status:MEDLINE


  8 / 9427 MEDLINE  
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[PMID]:29284028
[Au] Autor:McKenzie BA; Grover WH
[Ad] Endereço:Department of Bioengineering, Bourns College of Engineering, University of California Riverside, Riverside, CA 92521, United States of America.
[Ti] Título:A microfluidic thermometer: Precise temperature measurements in microliter- and nanoliter-scale volumes.
[So] Source:PLoS One;12(12):e0189430, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Measuring the temperature of a sample is a fundamental need in many biological and chemical processes. When the volume of the sample is on the microliter or nanoliter scale (e.g., cells, microorganisms, precious samples, or samples in microfluidic devices), accurate measurement of the sample temperature becomes challenging. In this work, we demonstrate a technique for accurately determining the temperature of microliter volumes using a simple 3D-printed microfluidic chip. We accomplish this by first filling "microfluidic thermometer" channels on the chip with substances with precisely known freezing/melting points. We then use a thermoelectric cooler to create a stable and linear temperature gradient along these channels within a measurement region on the chip. A custom software tool (available as online Supporting Information) is then used to find the locations of solid-liquid interfaces in the thermometer channels; these locations have known temperatures equal to the freezing/melting points of the substances in the channels. The software then uses the locations of these interfaces to calculate the temperature at any desired point within the measurement region. Using this approach, the temperature of any microliter-scale on-chip sample can be measured with an uncertainty of about a quarter of a degree Celsius. As a proof-of-concept, we use this technique to measure the unknown freezing point of a 50 microliter volume of solution and demonstrate its feasibility on a 400 nanoliter sample. Additionally, this technique can be used to measure the temperature of any on-chip sample, not just near-zero-Celsius freezing points. We demonstrate this by using an oil that solidifies near room temperature (coconut oil) in a microfluidic thermometer to measure on-chip temperatures well above zero Celsius. By providing a low-cost and simple way to accurately measure temperatures in small volumes, this technique should find applications in both research and educational laboratories.
[Mh] Termos MeSH primário: Técnicas Analíticas Microfluídicas/instrumentação
Temperatura Ambiente
Termômetros
[Mh] Termos MeSH secundário: Desenho de Equipamento
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, U.S. GOV'T, P.H.S.
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180129
[Lr] Data última revisão:
180129
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171229
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0189430


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[PMID]:29267312
[Au] Autor:Wang J; Rodgers VGJ; Brisk P; Grover WH
[Ad] Endereço:Department of Bioengineering, University of California Riverside, Riverside, CA, United States of America.
[Ti] Título:Instantaneous simulation of fluids and particles in complex microfluidic devices.
[So] Source:PLoS One;12(12):e0189429, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Microfluidics researchers are increasingly using computer simulation in many different aspects of their research. However, these simulations are often computationally intensive: simulating the behavior of a simple microfluidic chip can take hours to complete on typical computing hardware, and even powerful workstations can lack the computational capabilities needed to simulate more complex chips. This slows the development of new microfluidic chips for new applications. To address this issue, we present a microfluidic simulation method that can simulate the behavior of fluids and particles in some typical microfluidic chips instantaneously (in around one second). Our method decomposes the chip into its primary components: channels and intersections. The behavior of fluid in each channel is determined by leveraging analogies with electronic circuits, and the behavior of fluid and particles in each intersection is determined by querying a database containing nearly 100,000 pre-simulated channel intersections. While constructing this database takes a nontrivial amount of computation time, once built, this database can be queried to determine the behavior of fluids and particles in a given intersection in a fraction of a second. Using this approach, the behavior of a microfluidic chip can be simulated in just one second on a standard laptop computer, without any noticeable degradation in the accuracy of the simulation. While our current technique has some constraints on the designs of the chips it can simulate (namely, T- or cross-shaped intersections, 90 degree channel turns, a fixed channel width, fluid flow rates between 0 and 2 cm/s, and particles with diameters between 1 and 20 microns), we provide several strategies for increasing the range of possible chip designs that can be simulated using our technique. As a proof of concept, we show that our simulation method can instantaneously simulate the paths followed by particles in both simple and complex microfluidic chips, with results that are essentially indistinguishable from simulations that took hours or even days to complete using conventional approaches.
[Mh] Termos MeSH primário: Dispositivos Lab-On-A-Chip
Técnicas Analíticas Microfluídicas/instrumentação
[Mh] Termos MeSH secundário: Simulação por Computador
Desenho de Equipamento
Software
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, U.S. GOV'T, P.H.S.
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180116
[Lr] Data última revisão:
180116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171222
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0189429


  10 / 9427 MEDLINE  
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[PMID]:29266097
[Au] Autor:Okumus B; Baker CJ; Arias-Castro JC; Lai GC; Leoncini E; Bakshi S; Luro S; Landgraf D; Paulsson J
[Ad] Endereço:Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA.
[Ti] Título:Single-cell microscopy of suspension cultures using a microfluidics-assisted cell screening platform.
[So] Source:Nat Protoc;13(1):170-194, 2018 Jan.
[Is] ISSN:1750-2799
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Studies that rely on fluorescence imaging of nonadherent cells that are cultured in suspension, such as Escherichia coli, are often hampered by trade-offs that must be made between data throughput and imaging resolution. We developed a platform for microfluidics-assisted cell screening (MACS) that overcomes this trade-off by temporarily immobilizing suspension cells within a microfluidics chip. This enables high-throughput and automated single-cell microscopy for a wide range of cell types and sizes. As cells can be rapidly sampled directly from a suspension culture, MACS bypasses the need for sample preparation, and therefore allows measurements without perturbing the native cell physiology. The setup can also be integrated with complex growth chambers, and can be used to enrich or sort the imaged cells. Furthermore, MACS facilitates the visualization of individual cytoplasmic fluorescent proteins (FPs) in E. coli, allowing low-abundance proteins to be counted using standard total internal reflection fluorescence (TIRF) microscopy. Finally, MACS can be used to impart mechanical pressure for assessing the structural integrity of individual cells and their response to mechanical perturbations, or to make cells take up chemicals that otherwise would not pass through the membrane. This protocol describes the assembly of electronic control circuitry, the construction of liquid-handling components and the creation of the MACS microfluidics chip. The operation of MACS is described, and automation software is provided to integrate MACS control with image acquisition. Finally, we provide instructions for extending MACS using an external growth chamber (1 d) and for how to sort rare cells of interest.
[Mh] Termos MeSH primário: Técnicas de Cultura de Células/instrumentação
Técnicas Analíticas Microfluídicas/instrumentação
Microscopia/métodos
Análise de Célula Única/instrumentação
Análise de Célula Única/métodos
[Mh] Termos MeSH secundário: Desenho de Equipamento
Escherichia coli
Microscopia/instrumentação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180102
[Lr] Data última revisão:
180102
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171222
[St] Status:MEDLINE
[do] DOI:10.1038/nprot.2017.127



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