Trends in fluorescence imaging and related techniques to unravel biological information

Haustein, Elke; Schwille, Petra

doi:10.2976/1.2778852

Cited by 36 publications

(22 citation statements)

References 77 publications

(80 reference statements)

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“…The technical advances in fluorescence-based methods [42][43][44] and especially single-molecule fluorescence spectroscopy, including multicolour FRET [45] and high-resolution fluorescence microscopy, hold the promise to study transcription in living cells [46]. Furthermore, in concert with other singlemolecule techniques, such as atomic force microscopy and magnetic tweezers, short-lived intermediate states of the RNAP and transcription complexes can be isolated and characterized in great detail.…”

Section: Discussionmentioning

confidence: 99%

FRET (fluorescence resonance energy transfer) sheds light on transcription

Grohmann

Klose

Fielden

et al. 2011

Biochemical Society Transactions

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The complex organization of the transcription machinery has been revealed mainly by biochemical and crystallographic studies. X-ray structures describe RNA polymerases and transcription complexes on an atomic level, but fail to portray their dynamic nature. The use of fluorescence techniques has made it possible to add a new layer of information to our understanding of transcription by providing details about the structural rearrangement of mobile elements and the network of interactions within transcription complexes in solution and in real-time.

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Section: Discussionmentioning

confidence: 99%

FRET (fluorescence resonance energy transfer) sheds light on transcription

Grohmann

Klose

Fielden

et al. 2011

Biochemical Society Transactions

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“…The two main approaches to single-molecule fluorescence detection are confocal microscopy and widefield microscopy using the total internal reflection effect (Haustein and Schwille 2007;Moerner and Fromm 2003;Tinnefeld and Sauer 2005). In both cases, sensitivity on the single-molecule level is achieved by minimizing the excitation and detection volumes, either by introducing a confocal pinhole in the detection path (in confocal microscopy, Fig.…”

Section: Microscopy Apparatusmentioning

confidence: 99%

Single-molecule Förster resonance energy transfer studies of RNA structure, dynamics and function

2009

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Single-molecule fluorescence microscopy experiments on RNA molecules brought to light the highly complex dynamics of key biological processes, including RNA folding, catalysis of ribozymes, ligand sensing of riboswitches and aptamers, and protein synthesis in the ribosome. By using highly advanced biophysical spectroscopy techniques in combination with sophisticated biochemical synthesis approaches, molecular dynamics of individual RNA molecules can be observed in real time and under physiological conditions in unprecedented detail that cannot be achieved with bulk experiments. Here, we review recent advances in RNA folding and functional studies of RNA and RNA-protein complexes addressed by using single-molecule Förster (fluorescence) resonance energy transfer (smFRET) technique.

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“…Dentre as mais importantes há a microscopia de varredura confocal à laser, microscopia de fluorescência de 2 fótons, microscopia confocal com super-resolução óptica com depleção via emissão estimulada (STED), microscopia de fluorescên-cia em campo e com reflexão interna total (TIRF) entre outras. 66 As técnicas mais avançadas de microscopia de fluorescência possibilitam elucidar processos celulares em nível molecular. Com maior poder de resolução revelam, com resolução espacial e temporal, detalhes do processamento de nanomateriais por células eucarióticas.…”

Section: Figura 2 Local De Atuação De Inibidores De Endocitose Nas Vunclassified

“…Entre as vantagens adicionais podem ser citadas: técnica não destrutiva, menos laboriosa, menos dispendiosa, necessidade de uma menor quantidade de amostra e possibilidade de analisar amostras vivas. 66 A microscopia de fluorescência é fundamentada na emissão de luz por um fluoróforo endógeno ou exógeno da amostra e tem se tornado bastante comum, em virtude também da facilidade de manuseio do equipamento. Todavia, o preparo, marcação, análise e processamento das imagens devem ser feitos com o devido cuidado para não levar a interpretações equivocadas dos resultados.…”

Section: Figura 2 Local De Atuação De Inibidores De Endocitose Nas Vunclassified

Como estudar interações entre nanopartículas e sistemas biológicos

et al. 2016

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Recebido em 02/05/2016; aceito em 15/06/2016; publicado na web em 16/08/2016 STUDYING THE INTERACTIONS BETWEEN NANOPARTICLES AND BIOLOGICAL SYSTEMS. Although in recent years there has been an increasing amount of literature on nanotechnology and their clinical applications, it is still scarce a deep understanding of the interactions at the molecular levels between nanoparticles and cells. Studies demonstrating the underlying mechanisms of nanoparticles endocytosis, intracellular trafficking, and cellular processing are imperative to understand better how cells interact with those materials and their possible undesired effects, e.g. nanotoxicity. The rising awareness concerning nanoparticles applications and its interactions with the cellular environment is part of the new research field called Nanotoxicology. The cumulative knowledge in nanotoxicology will allow us to foresee toxic effects, establish regulations and limits for nanoparticles applications. In this work, we discuss the theoretical concepts about studying endocytosis and intracellular trafficking of nanoparticles. The nanoparticles-cell interactions are a multi-step process, which can be divided into nanoparticles' internalization, intracellular processing and triggering effects of nanomaterials on eukaryotic cells. Finally, we discuss the main techniques used to study this process: flow cytometry, use of endocytosis inhibitors and confocal microscopy.Keywords: endocytosis; intracellular trafficking; nanoparticles, nanotoxicology; nano-cell interactions. INTRODUÇÃONanotecnologia é um dos ramos mais promissores de tecnologia, sobre o qual se deposita grandes expectativas. 1 Somente em 2015, foram publicados cerca de 9400 artigos relacionados ao termo "Nanotechnology" no banco de dados Science Direct; além disso, diversos produtos têm sido testados e comercializados, por exemplo, protetores solares, cosméticos, produtos biomédicos, biosensores e sistemas para transporte e entrega de fármacos. 2 Entretanto, há uma crescente preocupação com a segurança e possíveis efeitos nocivos de nanomateriais que vêm sendo desenvolvidos, 3 pois dados e regulamentações internacionais referentes à segurança e uso desses materiais ainda são escassos, ainda estão em debate ou em fase de implementação. 4 Uma nova área da ciência destinada ao estudo de efeitos adversos de nanomateriais foi recentemente criada, a Nanotoxicologia. 5 O conhecimento acumulado nessa área tem como objetivo prever efeitos tóxicos e auxiliar nas regulamentações e diretrizes para aplicação da nanotecnologia 6 e uso de produtos a ela relacionados. A Figura 1 mostra o crescimento do número de artigos científicos que contêm o termo "nanotoxicology" e estão no banco de dados do National Center for Biotechnology Information (NCBI), subdivisão do National Institute of Health (NIH/USA).Apesar do número de artigos em nanotoxicologia ser crescente, esta é uma área ainda muito recente (com menos de uma década), e ainda não é possível determinar exatamente quais impactos os sistemas naturais e artificiais co...

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Trends in fluorescence imaging and related techniques to unravel biological information

Cited by 36 publications

References 77 publications

FRET (fluorescence resonance energy transfer) sheds light on transcription

FRET (fluorescence resonance energy transfer) sheds light on transcription

Single-molecule Förster resonance energy transfer studies of RNA structure, dynamics and function

Como estudar interações entre nanopartículas e sistemas biológicos

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