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Updated: Jun 24, 2026

Single Molecule Fluorescence Microscopy on Planar Supported Bilayers

Single Molecule Fluorescence Microscopy on Planar Supported Bilayers

Published on: October 31, 2015

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

Ruei-Yu He¹, Yuan-Deng Su, Keng-Chi Cho

¹Institute of Biomedical Engineering, National Cheng Kung University, Tainan 701, Taiwan.

|April 15, 2009

Summary

A new surface plasmon-enhanced two-photon total-internal-reflection fluorescence (TIRF) microscope offers brighter, clearer images of living cell membranes. This advanced TIRF microscopy reduces photobleaching and improves signal-to-noise ratio for enhanced cellular imaging.

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Area of Science:

Biophysics
Optical Microscopy
Cell Biology

Background:

Total-internal-reflection fluorescence (TIRF) microscopy is crucial for live-cell imaging.
Enhancing signal brightness and reducing photobleaching in TIRF remain challenges.
Two-photon excitation offers advantages in reduced scattering and smaller excitation volume.

Purpose of the Study:

To develop a novel surface plasmon-enhanced two-photon TIRF microscope.
To improve the quality of fluorescence imaging for living cell membranes.
To leverage surface plasmon (SP) effects for enhanced optical microscopy.

Main Methods:

Integration of surface plasmons (SPs) with two-photon excitation in a TIRF microscope setup.
Utilizing local electromagnetic field enhancement from SPs.
Employing two-photon excitation to reduce photobleaching and scattering.

Main Results:

Achieved significantly brighter fluorescence images of living cell membranes.
Demonstrated reduced photobleaching compared to conventional one-photon TIRF.
Obtained higher signal-to-noise ratio images due to smaller excitation volume and lower scattering.
Successfully imaged COS-7 fibroblasts transfected with EYFP-MEM or EGFP-WOX1.

Conclusions:

The developed surface plasmon-enhanced two-photon TIRF microscope provides superior contrast and brightness for live-cell membrane imaging.
This technique offers a powerful tool for studying dynamic cellular processes with reduced photodamage.
Combining SPs and two-photon excitation in TIRF microscopy significantly advances live-cell imaging capabilities.