Single-molecule spectroscopy of fluorescent proteins

Blum, Christian; Subramaniam, Vinod

doi:10.1007/s00216-008-2425-x

Cited by 33 publications

(31 citation statements)

References 138 publications

(229 reference statements)

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“…Howarth et al synthesized small quantum dots, having a hydrodynamic diameter of 11.1 ± 0.1 nm and a fluorescence quantum yield of ~0.4 17. Another class of fluorophores that are frequently used are fluorescent proteins, which have gained popularity as fluorescent labels in the last 10 years 18. Key to the success of fluorescent proteins (FP) is the easy introduction inside cells by genetic encoding of the FPs in vectors and plasmids, the non-toxicity and the specificity by fusing the FPs to proteins of interest.…”

Section: Introductionmentioning

confidence: 99%

Transfection of living HeLa cells with fluorescent poly-cytosine encapsulated Ag nanoclusters

Antoku

Hotta

Mizuno

et al. 2010

Photochem Photobiol Sci

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The fluorescence of silver clusters encapsulated by single stranded oligo-DNA (24 cytosine base pairs, C24:Agn) was used to monitor the transfection of this new silver/DNA fluorophore inside living HeLa cells. For this, the C24:Agn molecules were complexed with a commercially available transfection reagent Lipofectamine and the internalization of C24:Agn was followed with confocal fluorescence microscopy. Bright near-infrared fluorescence was observed from inside the transfected HeLa cells, when exciting with 633 nm excitation, opening up the possibility for the use of these C24:Agn clusters for biological labelling and imaging of living cells and for monitoring the transfection process with limited harm to the living cells.

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Section: Introductionmentioning

confidence: 99%

Transfection of living HeLa cells with fluorescent poly-cytosine encapsulated Ag nanoclusters

Antoku

Hotta

Mizuno

et al. 2010

Photochem Photobiol Sci

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“…Instruments already exist to measure for example fluorescence from a single protein molecule [161]. Such instrumentally smart technology of single-molecule spectroscopy enables the analysis of exactly one molecule at a time and could of course be a targeted or labeled molecule.…”

Section: Technologiesmentioning

confidence: 99%

Smart and green interfaces: From single bubbles/drops to industrial environmental and biomedical applications

Dutschk

Karapantsios

Liggieri

et al. 2014

Advances in Colloid and Interface Science

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Interfaces can be called Smart and Green (S&G) when tailored such that the required technologies can be implemented with high efficiency, adaptability and selectivity. At the same time they have also to be eco-friendly, i.e. products must be biodegradable, reusable or simply more durable. Bubble and drop interfaces are in many of these smart technologies the fundamental entities and help develop smart products of the everyday life.Significant improvements of these processes and products can be achieved by implementing and manipulating specific properties of these interfaces in a simple and smart way, in order to accomplish specific tasks. The severe environmental issues require in addition attributing ecofriendly features to these interfaces, by incorporating innovative , or, sometime, recycle materials and conceiving new production processes which minimize the use of natural resources and energy. Such concept can be extended to include important societal challenges related to support a sustainable development and a healthy population.The achievement of such ambitious targets requires the technology research to be supported by a robust development of theoretical and experimental tools, needed to understand in more details the behavior of complex interfaces. A wide but not exhaustive review of recent work concerned with green and smart interfaces is presented, addressing different scientific and technological fields. The presented approaches reveal a huge potential in relation to various technological fields, such as nanotechnologies, biotechnologies, medical diagnostics, new or improved materials.

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“…[29][30][31][32] For this reason, excitationmodulated measurements were also performed in the same manner on the fluorescent protein DsRed. In values obtained from these fits displayed a negative exponential dependence to  residence , (inset of figure 4).…”

Section: Experimental Verificationmentioning

confidence: 99%

Recovery of Photoinduced Reversible Dark States Utilized for Molecular Diffusion Measurements

2010

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For a spatially restricted excitation volume, the effective modulation of the excitation in time is influenced by the passage times of the molecules through the excitation volume. By applying an additional time-modulated excitation, the build-up of photo-induced reversible dark states in fluorescent molecules can be made to vary significantly with their passage times through the excitation volume. The variations in the dark state populations are reflected by the time-averaged fluorescence intensity, which thus can be used to characterize the mobilities of the molecules.The concept was experimentally verified by measuring the fluorescence response of freely diffusing Cyanine fluorophores (Cy5), undergoing trans-cis isomerisation when subject to time-modulated excitation in a focused laser beam. From the fluorescence response, and by applying a simple photodynamic model, the transition times of the Cy5 molecules could be well reproduced when applying different laminar flow speeds through the detection volume. The presented approach puts no constraints on sample concentration, no requirements for high time resolution or sensitivity in the detection, nor requires a high fluorescence brightness of the characterized molecules. This can make the concept useful for a broad range of biomolecular mobility studies.

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Single-molecule spectroscopy of fluorescent proteins

Cited by 33 publications

References 138 publications

Transfection of living HeLa cells with fluorescent poly-cytosine encapsulated Ag nanoclusters

Transfection of living HeLa cells with fluorescent poly-cytosine encapsulated Ag nanoclusters

Smart and green interfaces: From single bubbles/drops to industrial environmental and biomedical applications

Recovery of Photoinduced Reversible Dark States Utilized for Molecular Diffusion Measurements

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