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Updated: Aug 1, 2026
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Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
Published on: February 9, 2012
Compact multiphoton/single photon laser scanning microscope for spectral imaging and fluorescence lifetime imaging
Volker Ulrich1, Peter Fischer, Iris Riemann
1JenLab GmbH, Jena, Germany.
Scanning
|November 13, 2004
Summary
A compact laser scanning microscope (LSM) was enhanced for five-dimensional imaging, enabling detailed cellular and tissue fluorescence analysis with high spatial, temporal, and spectral resolution.
Area of Science:
- Biophotonics
- Microscopy
- Optical Imaging
Background:
- Advanced microscopy techniques are crucial for detailed biological sample analysis.
- Existing laser scanning microscopes (LSMs) can be limited in dimensionality and resolution.
- Developing compact, versatile imaging systems is essential for broader research applications.
Purpose of the Study:
- To upgrade an inverted fluorescence microscope into a compact, multi-dimensional laser scanning microscope (LSM).
- To integrate advanced imaging modalities including fluorescence lifetime imaging (FLIM) and spectral imaging.
- To investigate cellular and tissue fluorescence with high resolution using the developed system.
Main Methods:
- Modification of an inverted fluorescence microscope with a galvoscanner, z-stage, and picosecond laser.
- Incorporation of femtosecond lasers for multiphoton fluorescence and second harmonic generation (SHG) microscopy.
- Addition of time-correlated single photon counting and a Sagnac interferometer for FLIM and spectral imaging.
Main Results:
- Development of a compact five-dimensional microscope, TauMap (65 x 62 x 48 cm).
- Achieved submicron spatial resolution, <270 ps temporal resolution, and 10 nm spectral resolution.
- Successfully imaged single-photon excited (SPE) and two-photon excited (TPE) cellular fluorescence and plant tissue autofluorescence.
Conclusions:
- The developed TauMap system offers a unique platform for comprehensive 5D fluorescence imaging.
- The system enables high-resolution investigation of biological structures and processes.
- This compact, versatile microscope expands capabilities for biophotonics research.
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