Abstract
Metallic particles, silver in particular, can significantly enhance the fluorescence of dye molecules in the immediate vicinity (5-20 nm) of the particle. This magnifying effect can be theoretically explained/predicted by considering the change of photonic mode density near the fluorophore due to coupling to the conducting surface. We are using this method to observe fluorescence from a single ribosomal particle in a project aimed at acquiring sequence information from the translating ribosome (NIH's $1000 Genome Initiative). Several quartz slides with silver nanostructures were made using electron beam lithography techniques. The structures were approximately 50 nm high silver tiles measuring 400-700 nm on the side, and were spaced differently over a total area of 1 mm x 1 mm on any given quartz slide. In a preliminary experiment, we coated this surface with the Alexa 647-labeled antibodies and collected single molecule images using the MicroTime 200 (PicoQuant) confocal system. We showed that the fluorescence intensity measured over the silver islands film was more than 100-fold higher than fluorescence from a comparable site on uncoated section of the quartz slide. No noticeable photobleaching was seen. The fluorescence lifetime was very short, about 200 ps or less (this is the resolution limit of the system). The method has great promise for investigations of biologically relevant single molecules.
| Original language | English |
|---|---|
| Title of host publication | Single Molecule Spectroscopy and Imaging |
| Volume | 6862 |
| DOIs | |
| State | Published - 15 May 2008 |
| Event | Single Molecule Spectroscopy and Imaging - San Jose, CA, United States Duration: 19 Jan 2008 → 21 Jan 2008 |
Other
| Other | Single Molecule Spectroscopy and Imaging |
|---|---|
| Country | United States |
| City | San Jose, CA |
| Period | 19/01/08 → 21/01/08 |
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Keywords
- Confocal
- Fluorescence lifetime
- Microscopic
- Nanolithography
- Photobleaching
- Ribosome
- Silver islands
- Single molecule
Cite this
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Fluorescence enhancement on silver nanostructures : Studies of components of ribosomal translation in vitro. / Mandecki, Wlodek; Bharill, Shashank; Borejdo, Julian; Cabral, Diana; Cooperman, Barry S.; Farrell, Ian; Fetter, Linus; Goldman, Emanuel; Gryczynski, Zygmunt; Jakubowski, Hieronim; Liu, Hanqing; Luchowski, Rafal; Matveeva, Evgenia; Pan, Dongli; Qin, Haiou; Tennant, Donald; Gryczynski, Ignacy.
Single Molecule Spectroscopy and Imaging. Vol. 6862 2008. 68620T.Research output: Chapter in Book/Report/Conference proceeding › Conference contribution
TY - GEN
T1 - Fluorescence enhancement on silver nanostructures
T2 - Studies of components of ribosomal translation in vitro
AU - Mandecki, Wlodek
AU - Bharill, Shashank
AU - Borejdo, Julian
AU - Cabral, Diana
AU - Cooperman, Barry S.
AU - Farrell, Ian
AU - Fetter, Linus
AU - Goldman, Emanuel
AU - Gryczynski, Zygmunt
AU - Jakubowski, Hieronim
AU - Liu, Hanqing
AU - Luchowski, Rafal
AU - Matveeva, Evgenia
AU - Pan, Dongli
AU - Qin, Haiou
AU - Tennant, Donald
AU - Gryczynski, Ignacy
PY - 2008/5/15
Y1 - 2008/5/15
N2 - Metallic particles, silver in particular, can significantly enhance the fluorescence of dye molecules in the immediate vicinity (5-20 nm) of the particle. This magnifying effect can be theoretically explained/predicted by considering the change of photonic mode density near the fluorophore due to coupling to the conducting surface. We are using this method to observe fluorescence from a single ribosomal particle in a project aimed at acquiring sequence information from the translating ribosome (NIH's $1000 Genome Initiative). Several quartz slides with silver nanostructures were made using electron beam lithography techniques. The structures were approximately 50 nm high silver tiles measuring 400-700 nm on the side, and were spaced differently over a total area of 1 mm x 1 mm on any given quartz slide. In a preliminary experiment, we coated this surface with the Alexa 647-labeled antibodies and collected single molecule images using the MicroTime 200 (PicoQuant) confocal system. We showed that the fluorescence intensity measured over the silver islands film was more than 100-fold higher than fluorescence from a comparable site on uncoated section of the quartz slide. No noticeable photobleaching was seen. The fluorescence lifetime was very short, about 200 ps or less (this is the resolution limit of the system). The method has great promise for investigations of biologically relevant single molecules.
AB - Metallic particles, silver in particular, can significantly enhance the fluorescence of dye molecules in the immediate vicinity (5-20 nm) of the particle. This magnifying effect can be theoretically explained/predicted by considering the change of photonic mode density near the fluorophore due to coupling to the conducting surface. We are using this method to observe fluorescence from a single ribosomal particle in a project aimed at acquiring sequence information from the translating ribosome (NIH's $1000 Genome Initiative). Several quartz slides with silver nanostructures were made using electron beam lithography techniques. The structures were approximately 50 nm high silver tiles measuring 400-700 nm on the side, and were spaced differently over a total area of 1 mm x 1 mm on any given quartz slide. In a preliminary experiment, we coated this surface with the Alexa 647-labeled antibodies and collected single molecule images using the MicroTime 200 (PicoQuant) confocal system. We showed that the fluorescence intensity measured over the silver islands film was more than 100-fold higher than fluorescence from a comparable site on uncoated section of the quartz slide. No noticeable photobleaching was seen. The fluorescence lifetime was very short, about 200 ps or less (this is the resolution limit of the system). The method has great promise for investigations of biologically relevant single molecules.
KW - Confocal
KW - Fluorescence lifetime
KW - Microscopic
KW - Nanolithography
KW - Photobleaching
KW - Ribosome
KW - Silver islands
KW - Single molecule
UR - http://www.scopus.com/inward/record.url?scp=43249127839&partnerID=8YFLogxK
U2 - 10.1117/12.768612
DO - 10.1117/12.768612
M3 - Conference contribution
AN - SCOPUS:43249127839
SN - 9780819470379
VL - 6862
BT - Single Molecule Spectroscopy and Imaging
ER -