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Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy
Delphine Débarre1, Willy Supatto, Ana-Maria Pena
1Laboratory for optics and biosciences, Ecole Polytechnique, CNRS, INSERM, F-91128 Palaiseau, France.
Nature Methods
|December 22, 2005
Summary
Lipid bodies are key to energy storage and are highly visible using third-harmonic generation (THG) microscopy. This advanced imaging technique allows specific visualization of lipid bodies in unstained biological samples.
Area of Science:
- Cell Biology
- Biophysics
- Microscopy
Background:
- Lipid bodies are crucial for cellular energy storage and lipid metabolism.
- Current imaging methods often require staining, which can alter cellular structures and functions.
Purpose of the Study:
- To investigate the utility of third-harmonic generation (THG) microscopy for imaging lipid bodies.
- To demonstrate THG microscopy's capability in visualizing lipid bodies in various unstained biological samples.
Main Methods:
- Utilized third-harmonic generation (THG) microscopy with approximately 1,200 nm excitation.
- Applied epi-THG imaging to weakly absorbing tissues.
- Combined THG microscopy with two-photon and second-harmonic imaging.
Main Results:
- Lipid bodies generate a significantly strong THG signal in hepatocytes, enabling high-specificity imaging.
- THG microscopy successfully imaged lipid bodies in diverse unstained samples, including insect embryos, plant seeds, and mammalian tissues (liver, lung).
- Epi-THG imaging proved effective in weakly absorbing tissues due to efficient scattering of harmonic light.
Conclusions:
- Third-harmonic generation (THG) microscopy offers a powerful, label-free method for visualizing lipid bodies.
- This technique enhances our understanding of lipid body distribution and dynamics in biological systems.
- The multimodal imaging approach provides new insights into the complex interactions within tissues.