Long-term multiple color imaging of live cells using quantum dot bioconjugates

Jaiswal, Jyoti K.; Mattoussi, Hedi; Mauro, J. Matthew; Simon, Sanford M.

doi:10.1038/nbt767

Cited by 1,902 publications

(1,447 citation statements)

References 22 publications

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“…23,30 In contrast to Qdots with a nuclear localization sequence on the surface, PEGsilane-Qdots are unable to cross the nuclear membrane, 23 preventing their direct interaction with the genetic machinery in the cell nucleus. This precludes studies requiring the labeling of nuclear materials, creating a definite disadvantage.…”

Section: Discussionmentioning

confidence: 99%

Cellular Effect of High Doses of Silica-Coated Quantum Dot Profiled with High Throughput Gene Expression Analysis and High Content Cellomics Measurements

et al. 2006

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Quantum dots (Qdots) are now used extensively for labeling in biomedical research, and this use is predicted to grow because of their many advantages over alternative labeling methods. Uncoated Qdots made of core/shell CdSe/ZnS are toxic to cells because of the release of Cd 2+ ions into the cellular environment. This problem has been partially overcome by coating Qdots with polymers, poly(ethylene glycol) (PEG), or other inert molecules. The most promising coating to date, for reducing toxicity, appears to be PEG. When PEG-coated silanized Qdots (PEG-silane-Qdots) are used to treat cells, toxicity is not observed, even at dosages above 10-20 nM, a concentration inducing death when cells are treated with polymer or mercaptoacid coated Qdots. Because of the importance of Qdots in current and future biomedical and clinical applications, we believe it is essential to more completely understand and verify this negative global response from cells treated with PEG-silaneQdots. Consequently, we examined the molecular and cellular response of cells treated with two different dosages of PEG-silane-Qdots. Human fibroblasts were exposed to 8 and 80 nM of these Qdots, and both phenotypic as well as whole genome expression measurements were made. PEGsilane-Qdots did not induce any statistically significant cell cycle changes and minimal apoptosis/ necrosis in lung fibroblasts (IMR-90) as measured by high content image analysis, regardless of the treatment dosage. A slight increase in apoptosis/necrosis was observed in treated human skin fibroblasts (HSF-42) at both the low and the high dosages. We performed genome-wide expression array analysis of HSF-42 exposed to doses 8 and 80 nM to link the global cell response to a molecular and genetic phenotype. We used a gene array containing ~22,000 total probe sets, containing 18,400 probe sets from known genes. Only ~50 genes (~0.2% of all the genes tested) exhibited a statistically significant change in expression level of greater than 2-fold. Genes activated in treated cells included NIH Public Access

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Section: Discussionmentioning

confidence: 99%

Cellular Effect of High Doses of Silica-Coated Quantum Dot Profiled with High Throughput Gene Expression Analysis and High Content Cellomics Measurements

et al. 2006

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“…Employing  antibodies for surface modification has become a staple technique in the development of immunosensors (Virzonis et al 2014, Chrouda et al 2015, drug-delivery systems (Wu et al, Shahbazi et al 2014) and high-sensitivity biological labels (Jaiswal et al 2003, Park et al 2014. The simplest method of coupling antibodies to surfaces is physical adsorption.…”

Section: Introductionmentioning

confidence: 99%

Considerations in producing preferentially reduced half-antibody fragments

Makaraviciute

Jackson

Millner

et al. 2016

Journal of Immunological Methods

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Half-antibody fragments are a promising reagent for biosensing, drug-delivery and labeling applications, since exposure of the free thiol group in the Fc hinge region allows oriented Preferential reduction of rabbit anti-myoglobin IgG antibodies was optimized and the highest half-antibody yield was obtained with 35 mM TCEP. Finally, it has been demonstrated that produced anti-myoglobin half-IgG fragments retained their binding activity.

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“…In a detailed report, Jaiswal et al demonstrated passive endocytotic uptake of hydrophilic QDs capped with dihydrolipoic acid (DHLA) by incubating HeLa cells with solutions containing QDs at concentrations of 400 -600 nM QDs for 3-4 hours (1). QDs entering cells by this pathway remained largely sequestered within endocytic vesicles for several hours (1,11). Other methods for the cellular uptake of QDs that have been reported include: electroporation (14), cationic transfection reagents (11,15) and microinjection (11).…”

Section: Introductionmentioning

confidence: 99%

Self-Assembled Quantum Dot−Peptide Bioconjugates for Selective Intracellular Delivery

et al. 2006

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We demonstrate the use of self-assembled luminescent semiconductor quantum dot (QD) -peptide bioconjugates for the selective intracellular labeling of several eukaryotic cell lines. A bifunctional oligoarginine cell penetrating peptide (based on the HIV-1 Tat protein motif) bearing a terminal polyhistidine tract was synthesized and used to facilitate the transmembrane delivery of the QD bioconjugates. The polyhistidine sequence allows the peptide to self-assemble onto the QD surface via metal-affinity interactions while the oligoarginine sequence allows specific QD delivery across the cellular membrane and intracellular labeling as compared to non-conjugated QDs. This peptidedriven delivery is concentration-dependent and thus can be titrated. Upon internalization, QDs display a punctate-like staining pattern in which some, but not all, of the QD signal is co-localized within endosomes. The effects of constant versus limited exposure to QD-peptide conjugates on cellular viability are evaluated by a metabolic specific assay and clear differences in cytotoxicity are observed. The efficacy of using peptides for selective intracellular delivery is highlighted by performing a multicolor QD labeling, where we found that the presence or absence of peptide on the QD surface controls cellular uptake.

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Long-term multiple color imaging of live cells using quantum dot bioconjugates

Cited by 1,902 publications

References 22 publications

Cellular Effect of High Doses of Silica-Coated Quantum Dot Profiled with High Throughput Gene Expression Analysis and High Content Cellomics Measurements

Cellular Effect of High Doses of Silica-Coated Quantum Dot Profiled with High Throughput Gene Expression Analysis and High Content Cellomics Measurements

Considerations in producing preferentially reduced half-antibody fragments

Self-Assembled Quantum Dot−Peptide Bioconjugates for Selective Intracellular Delivery

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