Probing Single Molecules and Single Nanoparticles by Surface-Enhanced Raman Scattering
Tóm tắt
Từ khóa
Tài liệu tham khảo
Xie X. S. and , Dunn R. C., ibid. 265, 361 (1994).
Nie S. and , Zare R. N., Annu. Rev. Biophys. Biomol. Struct., in press.
Schmidt Th. Schutz G. J. Baumgartner W. Gruber H. J. Schindler H. Proc. Natl. Acad. Sci. U.S.A. 93 2926 (1996);
Haab B. B. and , Mathies R. A., ibid. 67, 3253 (1995).
Collinson M. M. and , Wightman R. M., ibid. 268, 1883 (1995).
Kneipp K. et al. abstract 24 presented at the 23rd Annual Conference of the Federation of Analytical Chemistry and Spectroscopy Societies Kansas City MO 29 September to 4 October 1996.
A 250-μl aliquot of Ag colloid prepared by the procedure of P. C. Lee and D. Meisel [ J. Phys. Chem. 86 3391 (1982)] was incubated with R6G and ∼1 mM NaCl in a 1.5-ml plastic microcentrifuge tube. Glass containers should be avoided because of R6G adsorption on glass surfaces. At the electrolyte concentrations used the Ag colloid was activated but not aggregated after an extended incubation time of ∼3 hours at room temperature. The amount of free R6G in solution was experimentally determined to be ∼20% by centrifugation of the Ag particles and fluorescence measurement of the supernatant solution. This result is in agreement with the calculated value (∼10 to 20%) based on the equilibrium binding constant 1.8 × 10 −9 M (19). Bulk R6G concentrations from 10 −7 to 10 −11 M were calibrated by fluorescence measurement in the absence of Ag colloids. The order speed and buffer volume of the colloid-analyte mixing process did not make measurable differences indicating that equilibrium adsorption conditions were reached in our experiment.
In this integrated microscope ultrasensitive optical imaging and spectroscopy provided molecular information and AFM resolved the shape and size of individual nanoparticles. The microscope side port was coupled to a high-throughput single-stage spectrograph and a back-illuminated charge-coupled device (CCD) detector for spectroscopy. The microscope front was attached to a video-rate intensified CCD for wide-field imaging of single nanoparticles with epi- or evanescent-wave laser excitation. A tapping-mode AFM scanning head was mounted directly on the microscope stage for topographic imaging at nanometer-scale resolution. When coupled with a video data acquisition system this apparatus allowed digital movies to be made of single nanoparticles at 30 frames per second. Detailed instrument diagram and specifications are available upon request.
This question can be addressed by using near-field scanning optical microscopy to image nanoparticle aggregates at a resolution of 50 to 100 nm. Recent research shows that both surface and resonance-enhanced Raman spectra can be obtained with a near-field fiber probe in nanometer domains [S. R. Emory and S. Nie in International Conference on Raman Spectroscopy S. A. Asher and P. B. Stein Eds. (Wiley New York 1996) pp. 1176–1177; S. Webster D. A. M. Smith M. W. Ayad K. Kershaw D. N. Batchelder ibid. pp. 1146–1147].
Jakobi H., Kuhn H., Ber. Bunsenges. Phys. Chem. 66, 46 (1962).
Golab J. T., Sprague J. R., Carron K. T., Schatz G. C., Van Duyne R. P., J. Chem. Phys. 88, 7942 (1988).
ibid. 98, 11751 (1994).
Xiao T. Ye Q. Sun L. J. Phys. Chem. in press.
S.N. acknowledges the Whitaker Foundation for a Biomedical Engineering Award and the Beckman Foundation for a Beckman Young Investigator Award. This work was supported by Indiana University Startup Funds.