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Volumn 281, Issue 5385, 1998, Pages 2013-2016

Semiconductor nanocrystals as fluorescent biological labels

Author keywords

[No Author keywords available]

Indexed keywords

EMISSION SPECTROSCOPY; FLUORESCENCE; PROBES; SEMICONDUCTOR MATERIALS;

EID: 0012392952     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.281.5385.2013     Document Type: Article
Times cited : (8546)

References (30)
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    • Attempts with energy transfer-sensitized probes and phycobiliprotein-conjugated dye molecules have been somewhat successful, but the efficiency of energy transfer is lower than the intrinsic excitation and emission of a dye molecule [J. Y. Ju, C. C. Ruan, C. W. Fuller, A. N. Glazer, R. A. Mathies, Proc. Natl. Acad. Sci. U.S.A. 92, 4347 (1995); M. Roederer, A. B. Kantor, D. R. Parks, L. A. Herzenberg, Cytometry 24, 191 (1996)].
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    • The idea of using biological interactions to pattern materials has also been explored [A. P. Alivisatos et al., Nature 382, 609 (1996); C. A. Mirkin, R. L. Letsinger, R. C. Mucic, J. J. Storhoff, ibid., p. 607].
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    • The idea of using biological interactions to pattern materials has also been explored [A. P. Alivisatos et al., Nature 382, 609 (1996); C. A. Mirkin, R. L. Letsinger, R. C. Mucic, J. J. Storhoff, ibid., p. 607].
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    • note
    • 2O, was added, the preparation stirred for another 2 hours, then concentrated partially at 60°C in vacuo to an oily solution. This solution was precipitated to a greasy solid with 50% acetone in isopropanol. This solid was redissolved in water and buffered solution.
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    • note
    • For conversion of functional groups on the surface of the nanocrystals, the precipitated sample was dissolved in 100 mM phosphate-buffered saline (PBS), pH 7.5, at a concentration four times that of the final oily solution. A 1.0-ml volume of the concentrated nanocrystal solution was incubated with 100 to 400 μl of a 10-mg/ml solution of biotinamidocaproic acid 3-sulfo-N-hydroxysuccinimide ester (Sigma) in 6% DMSO in PBS for 1 hour, after which the solution was quenched with a neutralized iodoacetic acid solution in PBS to 50 mM final iodoacetate concentration. The unbiotinylated nanocrystals were prepared by adding the quench solution directly. These solutions were incubated overnight at 4°C, then concentrated and rinsed through a Centricon 50 (Millipore) and diluted to 1.0 ml; the remaining amines on the biotinylated nanocrystals were capped with 50 μl of a 1 M solution of succinic anhydride in dry DMSO, followed by addition of an equivalent amount of base and further purification through a Centricon 50. The retentate was diluted to 1.0 ml and used for labeling at 1:10 dilution. The biotinylated nanocrystals shifted to lower mobility in a gel when incubated with streptavidin. Higher levels of biotinylation followed by streptavidin incubation resulted in a further decrease in mobility through agarose gel. These effects were not seen in unbiotinylated nanocrystal samples, indicating that this interaction is specific and that the biotin is covalently bound to the nanocrystal surface.
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    • -1. In this report, we refer to this emission as the nanocrystal fluorescence [M. Nirmal et al., Phys. Rev. Lett. 75, 3728 (1995)].
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    • note
    • Mouse 3T3 fibroblasts were grown on fibronectin-treated, formvar-coated gold grids and fixed with 4% paraformaldehyde, 0.1% glutaraldehyde, and 0.5% Triton at room temperature. Specimens were then treated successively with unbiotinylated green nanocrystals, phalloidin-biotin (Molecular Probes) (15 min), streptavidin (25 μg/ml) (30 min), a ∼100 nM solution of biotinylated red nanocrystals (30 min), again streptavidin (30 min), then once more with the red biotinylated nanocrystals (30 min). At least three PBS or Superblock-PBS (Pierce) rinse steps (5 min each) were performed between each incubation. Samples were mounted on microscope slides in PBS.
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    • note
    • We would like to thank X. Peng, J. Gray, C. Bertozzi, and P. Schultz for many helpful discussions. The photoluminescence spectra of InAs nanocrystals, InP nanocrystals, and CdSe nanocrystals in Fig. 2 were provided by J. Wickham, U. Banin, and X. Peng. Supported by the Director, Office of Energy Research, Office of Basic Energy Sciences, Division of Materials Sciences, of the U.S. Department of Energy under contract number DEAC03-76SF00098. M.B. acknowledges support from a NSF graduate research fellowship.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.