Single molecule immunoassay on plasmonic platforms.
We examined the photophysical properties of the new near infrared (NIR) fluorescent label SeTau-665 on a plasmonic platform of self- assembled colloidal structures (SACS) of silver prepared on a semitransparent silver film. A SeTau-665 immunoassay was performed on this platform and a control glass slide. The fluorescence properties of this label substantially change due to plasmonic interactions. While the average brightness increase of SeTau 665 in ensemble measurements was about 70-fold, fluorescence enhancements up to four-hundred times were observed on certain "hot spots" for single molecule measurements. The intensity increase is strongly correlated with a simultaneous decrease in fluorescence lifetime in these "hot spots". The large increase in brightness allows the reduction of the excitation power resulting in a reduced background and increased photostability. The remarkable fluorescence enhancements observed for SeTau 665 on our plasmonic platform should allow to substantially improve single molecule detection and to reduce the detection limits in sensing devices.
Luchowski
R
R
Center for Commercialization of Fluorescence Technologies, Department of Molecular Biology & Immunology, UNTHSC, Fort Worth, TX 76107, USA. rluchows@hsc.unt.edu
Matveeva
E G
EG
Shtoyko
T
T
Sarkar
P
P
Patsenker
L D
LD
Klochko
O P
OP
Terpetschnig
E A
EA
Borejdo
J
J
Akopova
I
I
Gryczynski
Z
Z
Gryczynski
I
I
D015374:Biosensing Techniques / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007118:Immunoassay / Q000295:instrumentation*; D020349:Surface Plasmon Resonance / Q000295:instrumentation*