Journal
JOURNAL OF LUMINESCENCE
Volume 98, Issue 1-4, Pages 81-89Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S0022-2313(02)00255-7
Keywords
single molecule imaging; surface-enhanced Raman scattering; fluorescence; adenine; TNP-ATP; Klenow fragment; silver coloidal particles
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We implemented Raman imaging and spectroscopy of adenine molecules adsorbed on Ag colloidal nanoparticles to find feasibility of detecting and identifying nucleic-acid bases at the single-molecule level. Surface enhanced Raman scattering of adenine molecules showed an intermittent on-and-off behavior called blinking and fluctuation of the center of spectra called spectral diffusion. The observation of blinking and spectral diffusion provides substantial evidence for detecting single adenine molecules. Also, we extended single molecule fluorescence imaging and time-resolved fluorometry from green to violet-excitation regime to detect a fluorescent analog of nucleic-acid bases at the single-molecule level. Using violet excitation we observed fluorescence spectra and lifetimes from single complexes of an analog of adenine and the Klenow fragment of DNA polymerase I. (C) 2002 Published by Elsevier Science B.V.
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