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Volumn 288, Issue 5464, 2000, Pages 316-318

Translating biomolecular recognition into nanomechanics

Author keywords

[No Author keywords available]

Indexed keywords

ANTISENSE OLIGONUCLEOTIDE; IMMUNOGLOBULIN; NANOPARTICLE; PROTEIN A;

EID: 0034646725     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.288.5464.316     Document Type: Article
Times cited : (1609)

References (26)
  • 9
    • 0030769421 scopus 로고    scopus 로고
    • R. Berger et al., Science 276, 2021 (1997).
    • (1997) Science , vol.276 , pp. 2021
    • Berger, R.1
  • 13
    • 0343354396 scopus 로고    scopus 로고
    • note
    • The liquid cell has a volume of 300 μl, and the buffer solution was manually exchanged by a micropipette. All hybridization experiments were performed in saline sodium citrate hybridization buffer (HB) (5X SSC buffer, Sigma Chemicals) at room temperature (22.0° ± 0.2°C).
  • 14
    • 0001022448 scopus 로고    scopus 로고
    • Each cantilever is illuminated by one vertical cavity surface-emitting laser (VCSEL; wavelength 760 nm; CSEM, Zurich, Switzerland) of a linear VCSEL array (pitch 250 μm). The VCSELs are time-multiplexed with a rate of 1.3 Hz. Detection of the reflected light by a single linear position-sensitive detector (Sitek, Partille, Sweden) was used to measure the bending of each cantilever. The cantilever deflection is calculated with an accuracy of 0.1 nm [T. Miyatani and M. Fujihira, J. Appl. Phys. 81, 7099 (1997)].
    • (1997) J. Appl. Phys. , vol.81 , pp. 7099
    • Miyatani, T.1    Fujihira, M.2
  • 15
    • 0343354395 scopus 로고    scopus 로고
    • note
    • Oligonucleotides 5′-functionalized by a hexyl spacer with a thiol group and purified twice by high-pressure liquid chromatography (HPLC) were used as obtained from Microsynth GmbH (Belgach, Switzerland). The thiol modification enables covalent binding to gold surfaces. Two cantilevers, each homogeneously covered on one side with a 20-nm-thick gold layer, were inserted in parallel for 20 min into two separate reservoirs. The two reservoirs were filled with 10 μl of a 40 μM solution of two different oligonucleotides, each in a 50 mM triethylammonium acetate buffer containing 25% ethanol. Functionalized arrays can be stored for several days at 4°C in air without loss of performance. Before use, arrays were equilibrated for several hours in the liquid cell.
  • 16
    • 0342484976 scopus 로고    scopus 로고
    • note
    • The resonance frequencies of eight functionalized cantilevers of one array vary by only 2%.
  • 20
    • 0342919190 scopus 로고    scopus 로고
    • note
    • 2 per oligonucleotide on gold was obtained, consistent with an averaged thickness of the oligonucleotide layer of ∼1.5 nm determined by ellipsometry and AFM. AFM also indicated a homogeneous monolayer.
  • 23
    • 0343354359 scopus 로고    scopus 로고
    • note
    • The gold surface of two cantilevers was blocked for protein adsorption by a layer of 11 -(ω-methoxypentaethylene glycol)undecan-1-thiol (courtesy of F. Kamounah, University of Copenhagen, Denmark). Protein A from Staphylococcus aureus [20 μg/ml in phosphate-buffered saline (PBS) (pH 7.4), Sigma Chemicals] was adsorbed on the silicon side of one cantilever for 1 hour. Then, the second cantilever together with the entire array was saturated with BSA for 1 hour (1 mg/ml in PBS, Sigma Chemicals). Rabbit IgG (0.1 mg/ml in PBS, Sigma Chemicals) and goat IgG (0.1 mg/ml in PBS, Sigma Chemicals) were used. All experiments were done in PBS at room temperature. Adsorption and block of proteins was checked by ellipsometry and AFM.
  • 26
    • 0342919157 scopus 로고    scopus 로고
    • note
    • We acknowledge the experimental support of M. Despont, U. Drechsler, R. Widmer, the continuous support of P. Seidler, and useful discussions with E. Delamarche and A. Bernard. This project was partially funded through the Swiss Priority Program Micro- and Nanosystem Technology (MINAST).


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.