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Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
Published on: February 9, 2012
Fluorescence lifetime imaging with pulsed diode laser enabled stimulated emission
Jianhong Ge1, Cuifang Kuang, Shin-Shian Lee
1State Key Laboratory of Modern Optical Instrumentation, Zhejiang University, Hangzhou, 310027, China. jianhong@zju.edu.cn
Optics Express
|December 25, 2012
Summary
This study introduces a new fluorescence lifetime imaging (FLIM) method using synchronized diode lasers for excitation and stimulated emission. This cost-effective, all-semiconductor approach enables high-performance FLIM at long working distances.
Area of Science:
- Optics and Photonics
- Biophotonics
- Laser Technology
Background:
- Fluorescence Lifetime Imaging (FLIM) is a powerful microscopy technique.
- Conventional FLIM systems can be complex and expensive.
- Stimulated emission offers potential for enhanced imaging capabilities.
Purpose of the Study:
- To develop a novel, cost-effective FLIM scheme.
- To utilize stimulated emission for improved imaging performance.
- To create a compact, all-semiconductor FLIM module.
Main Methods:
- Employed a pair of synchronized, gain-switched diode lasers (635 nm and 700 nm).
- Used lasers as excitation and stimulation light sources for ATTO647N labeled samples.
- Controlled relative time delay electronically for FLIM acquisition.
- Leveraged the coherent nature of stimulated emission signals.
Main Results:
- Successfully demonstrated stimulated emission-based FLIM.
- Achieved FLIM by electronically controlling laser time delays.
- Enabled long working distance FLIM due to coherent stimulated emission.
- Realized a high-performance, all-semiconductor FLIM module.
Conclusions:
- The presented scheme offers a flexible, compact, and cost-effective FLIM solution.
- This all-semiconductor approach simplifies FLIM instrumentation.
- The technique is suitable for applications requiring long working distance imaging.
