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 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.
Institute of Biomedical Engineering, National Cheng Kung University, Tainan 701, Taiwan.
Ruei-Yu
RY
He
Yuan-Deng
YD
Su
Keng-Chi
KC
Cho
Chun-Yun
CY
Lin
Nan-Shan
NS
Chang
Chih-Han
CH
Chang
Shean-Jen
SJ
Chen
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007089:Image Enhancement / Q000295:instrumentation*; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000295:instrumentation*