Surface plasmon-enhanced two-photon fluorescence microscopy for live cell membrane imaging

A surface plasmon-enhanced two-photon total-internal-reflection fluorescence (TIRF) microscope has been developed to provide fluorescent images of living cell membranes. The proposed microscope with the help of surface plasmons (SPs) not only provides brighter fluorescent images based on the mechani...

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Bibliographic Details
Published in:Optics express 2009-04, Vol.17 (8), p.5987-5997
Main Authors: He, Ruei-Yu, Su, Yuan-Deng, Cho, Keng-Chi, Lin, Chun-Yun, Chang, Nan-Shan, Chang, Chih-Han, Chen, Shean-Jen
Format: Article
Language:English
Subjects:
Animals
Cercopithecus aethiops
COS Cells
Equipment Design
Equipment Failure Analysis
Image Enhancement - instrumentation
Microscopy, Fluorescence, Multiphoton - instrumentation
Reproducibility of Results
Sensitivity and Specificity
Surface Plasmon Resonance - instrumentation
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Summary:A surface plasmon-enhanced two-photon total-internal-reflection fluorescence (TIRF) microscope has been developed to provide fluorescent images of living cell membranes. The proposed microscope with the help of surface plasmons (SPs) not only provides brighter fluorescent images based on the mechanism of local electromagnetic field enhancement, but also reduces photobleaching due to having a shorter fluorophore lifetime. In comparison with a one-photon TIRF, the two-photon TIRF can achieve higher signal-to-noise ratio cell membrane imaging due its smaller excitation volume and lower scattering. By combining the SP enhancement and two-photon excitation TIRF, the microscope has demonstrated it's capability for brighter and more contrasted fluorescence membrane images of living monkey kidney COS-7 fibroblasts transfected with an EYFP-MEM or EGFP-WOX1 construct.
ISSN:1094-4087
1094-4087
DOI:10.1364/oe.17.005987