Single-Step Multicolor Fluorescence In Situ Hybridization Using Semiconductor Quantum Dot-DNA Conjugates

Bentolila, Laurent A.; Weiss, Shimon

doi:10.1385/cbb:45:1:59

Cited by 57 publications

(39 citation statements)

References 25 publications

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“…Xiao and Barker demonstrated increased photostability of QD based probes versus conventional dyes for the probing of human metaphase chromosomes (see Figure 5). Chan and colleagues, and Bentolila and Weiss also demonstrated increased sensitivity and the ability to multiplex in these assay formats (Chan et al 2005;Bentolila and Weiss 2006). These results indicate that the relatively large QD-DNA probe complexes can still penetrate the tight cellular and chromosomal structures while retaining hybridization specifi city.…”

Section: Nucleic Acid Detectionmentioning

confidence: 88%

Potential clinical applications of quantum dots

Clapp¹

2008

IJN

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Abstract:The use of luminescent colloidal quantum dots in biological investigations has increased dramatically over the past several years due to their unique size-dependent optical properties and recent advances in biofunctionalization. In this review, we describe the methods for generating high-quality nanocrystals and report on current and potential uses of these versatile materials. Numerous examples are provided in several key areas including cell labeling, biosensing, in vivo imaging, bimodal magnetic-luminescent imaging, and diagnostics. We also explore toxicity issues surrounding these materials and speculate about the future uses of quantum dots in a clinical setting.

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Section: Nucleic Acid Detectionmentioning

confidence: 88%

Potential clinical applications of quantum dots

Clapp¹

2008

IJN

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“…That is, while several studies have demonstrated the use of QDs in FISH, as with the majority of studies in the literature, there may be a tendency to present only the positive data. QDbased FISH studies are conspicuous mostly by their absence (Xiao and Barker 2004a;Bentolila and Weiss 2006;Maetal.2008); that is, if QDs had fulfilled their promise they would, at least in part, have replaced organic fluorochromes. One would expect orders of magnitude more QD-FISH papers in the literature and several companies marketing QD-labelled probes, which-at the time of writing-is simply not the case.…”

Section: Discussionmentioning

confidence: 99%

“…Biotinylated labelled oligonucleotides were conjugated with QDstreptavidin in the presence of biocytin to block excess streptavidin sites that could result in oligonucleotide cross-linking. Bentolila and Weiss (2006) using a biotin-streptavidin strategy, labelled oligonucleotide probes with QDs; in this case complexes were analysed using gel electrophoresis and the optimum molar ratio of QD-DNA was used against the major (γ) family of mouse satellite DNA in both interphase and metaphase preparations. In addition they also used oligonucleotides labelled with different coloured QDs to target two classes of repetitive DNA in the centromeric region.…”

Section: Introductionmentioning

confidence: 99%

Quantum dots as new-generation fluorochromes for FISH: an appraisal

et al. 2009

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In the field of nanotechnology, quantum dots (QDs) are a novel class of inorganic fluorochromes composed of nanometre-scale crystals made of a semiconductor material. Given the remarkable optical properties that they possess, they have been proposed as an ideal material for use in fluorescent in-situ hybridization (FISH). That is, they are resistant to photobleaching and they excite at a wide range of wavelengths but emit light in a very narrow band that can be controlled by particle size and thus have the potential for multiplexing experiments. The principal aim of this study was to compare the potential of QDs against traditional organic fluorochromes in both indirect (i.e. QD-conjugated streptavidin) and direct (i.e. synthesis of QD-labelled FISH probes) detection methods. In general, the indirect experiments met with a degree of success, with FISH applications demonstrated for chromosome painting, BAC mapping and use of oligonucleotide probes on human and avian chromosomes/nuclei. Many of the reported properties of QDs (e.g. brightness, 'blinking' and resistance to photobleaching) were observed. On the other hand, signals were more frequently observed where the chromatin was less condensed (e.g. around the periphery of the chromosome or in the interphase nucleus) and significant bleed-through to other filters was apparent (despite the reported narrow emission spectra). Most importantly, experimental success was intermittent (sometimes even in identical, parallel experiments) making attempts to improve reliability difficult. Experimentation with direct labelling showed evidence of the generation of QD-DNA constructs but no successful FISH experiments. We conclude that QDs are not, in their current form, suitable materials for FISH because of the lack of reproducibility of the experiments; we speculate why this might be the case and look forward to the possibility of nanotechnology forming the basis of future molecular cytogenetic applications.

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“…A rapid method for the direct multicolor imaging of multiple subnuclear genetic sequences uses novel QD-based FISH. A Texas red dye gamma-satellite probe produces fluorescent foci at the periphery of interphase nucleus and labels every centromere in metaphase chromosomes (9 ).…”

Section: Nanotechnology-based Cytogeneticsmentioning

confidence: 99%

Applications of Nanobiotechnology in Clinical Diagnostics

2007

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Background: Nanobiotechnologies are being applied to molecular diagnostics and several technologies are in development. Methods: This review describes nanobiotechnologies that are already incorporated in molecular diagnostics or have potential applications in clinical diagnosis. Selected promising technologies from published literature as well as some technologies that are in commercial development but have not been reported are included. Results: Nanotechnologies enable diagnosis at the single-cell and molecule levels, and some can be incorporated in current molecular diagnostic methods, such as biochips. Nanoparticles, such as gold nanoparticles and quantum dots, are the most widely used, but various other nanotechnological devices for manipulation at the nanoscale as well as nanobiosensors are also promising for potential clinical applications. Conclusions: Nanotechnologies will extend the limits of current molecular diagnostics and enable point-ofcare diagnostics, integration of diagnostics with therapeutics, and development of personalized medicine. Although the potential diagnostic applications are unlimited, the most important current applications are foreseen in the areas of biomarker discovery, cancer diagnosis, and detection of infectious microorganisms. Safety studies are needed for in vivo use. Because of its close interrelationships with other technologies, nanobiotechnology in clinical diagnosis will play an important role in the development of nanomedicine in the future.

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Single-Step Multicolor Fluorescence In Situ Hybridization Using Semiconductor Quantum Dot-DNA Conjugates

Cited by 57 publications

References 25 publications

Potential clinical applications of quantum dots

Potential clinical applications of quantum dots

Quantum dots as new-generation fluorochromes for FISH: an appraisal

Applications of Nanobiotechnology in Clinical Diagnostics

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