Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy
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- 20 December 2005
- journal article
- Published by Springer Nature in Nature Methods
- Vol. 3 (1) , 47-53
- https://doi.org/10.1038/nmeth813
Abstract
Lipid bodies have an important role in energy storage and lipid regulation. Here we show that lipid bodies are a major source of contrast in third-harmonic generation (THG) microscopy of cells and tissues. In hepatocytes, micrometer-sized lipid bodies produce a THG signal 1-2 orders of magnitude larger than other structures, which allows one to image them with high specificity. THG microscopy with approximately 1,200 nm excitation can be used to follow the distribution of lipid bodies in a variety of unstained samples including insect embryos, plant seeds and intact mammalian tissue (liver, lung). We found that epi-THG imaging is possible in weakly absorbing tissues because bulk scattering redirects a substantial fraction of the forward-generated harmonic light toward the objective. Finally, we show that the combination of THG microscopy with two-photon and second-harmonic imaging provides a new tool for exploring the interactions between lipid bodies, extracellular matrix and fluorescent compounds (vitamin A, NADH and others) in tissues.Keywords
This publication has 32 references indexed in Scilit:
- Caveolin, cholesterol, and lipid bodiesSeminars in Cell & Developmental Biology, 2005
- Intracellular origin and secretion of milk fat globulesEuropean Journal of Cell Biology, 2005
- RetinosomesThe Journal of cell biology, 2004
- Nonlinear magic: multiphoton microscopy in the biosciencesNature Biotechnology, 2003
- Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generationProceedings of the National Academy of Sciences, 2003
- The biogenesis and functions of lipid bodies in animals, plants and microorganismsProgress in Lipid Research, 2001
- Mechanisms of lipid-body formationTrends in Biochemical Sciences, 1999
- Third harmonic generation microscopyOptics Express, 1998
- Nonlinear scanning laser microscopy by third harmonic generationApplied Physics Letters, 1997
- Nile red: a selective fluorescent stain for intracellular lipid droplets.The Journal of cell biology, 1985