NDLI: Prism-Based Spectral Imaging of Single-Molecule Fluorescence from Gold-Nanoparticle/Fluorophore Complex

Content Provider	Springer Nature Link
Author	Sonehara, Tsuyoshi Sakai, Tomoyuki Haga, Takabu Fujita, Takeshi Takahashi, Satoshi
Copyright Year	2011
Abstract	A wavelength-calibration method for prism-based spectral imaging of single-molecule (SM) fluorescence was developed. With this method, a wavelength reference is provided by photoluminescence from 50-nm-diameter gold nanoparticles (AuNPs) binding with fluorophores. The AuNPs each bound with a SM fluorophore, either Alexa488 or Cy3, to form AuNP/fluorophore complexes in tris-HCl buffer. Each complex was immobilized on a silica slide and then excited by total-internal-reflection illumination to make it emit SM fluorescence and AuNP photoluminescence. The portion of the AuNP photoluminescence transmitted by a band-pass filter gives the wavelength reference. A spectral-imaging system composed of a prism-based spectroscope (with a reciprocal dispersion of about 4 nm/μm) and a charge-coupled device with 6.45-μm-square pixels was used to obtain an SM-fluorescence spectrum and a wavelength-reference spectrum. Through smoothed differentiation of these two spectra, the peak location of the former in relation to the latter was determined with subpixel precision. After that, the SM fluorophore was classified as either Alexa488 or Cy3 according to the peak location. The error rate of the classification was estimated to be only 0.3%.
Starting Page	1805
Ending Page	1811
Page Count	7
File Format	PDF
ISSN	10530509
Journal	Journal of Fluorescence
Volume Number	21
Issue Number	4
e-ISSN	15734994
Language	English
Publisher	Springer US
Publisher Date	2011-03-08
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Wavelength calibration Prism Single molecule Gold nanoparticle Photoluminescence Biochemistry Biotechnology Biophysics and Biological Physics Biomedicine general Analytical Chemistry
Content Type	Text
Resource Type	Article
Subject	Sociology and Political Science Spectroscopy Law Biochemistry Clinical Biochemistry Clinical Psychology

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3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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