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CARS MICROSCOPYCoherent anti-Stokes Raman Scattering (CARS) microscopy allows rapid and non-perturbative chemical imaging particularly of lipids in living cells. Visualization in CARS microscopy arises from the intrinsic vibrations of molecules.
The technology was developed in the lab of Xiaoliang Sunney Xie, Professor of Chemistry and Chemical Biology at Harvard University (http://bernstein.harvard.edu/research/cars.html). Leica Microsystems and Harvard Universitys Office of Technology Development (OTD) recently announced a license agreement to develop CARS microscopy as an extension of confocal/multiphoton microscopes (http://www.leica-microsystems.com).
The IMB Microscopy Facility was worldwide the first user of a commercial CARS microscope!
Wolinski, H., Kohlwein, S.D. Microscopic and spectroscopic techniques to investigate lipid droplet formation and turnover in yeast (2015) Methods in Molecular Biology, 1270, pp. 289-305.
Margret Paar, Christian Juengst, Noemi A. Steiner, Christoph Magnes, Frank Sinner, Dagmar Kolb, Achim Lass, Robert Zimmermann, Andreas Zumbusch, Sepp D. Kohlwein, and Heimo Wolinski. 2012. Remodeling of lipid droplets during lipolysis and growth in adipocytes. J. Biol. Chem. published February 6, 2012 as doi:10.1074/jbc.M111.316794
Wolinski H, Bredies, K, Sepp D. Kohlwein. 2012. Quantitative imaging of lipid metabolism in yeast: from 4D analysis to high content screens of mutant libraries.Meth Cell Biol. 108:345-65.
Visualization of Perilipin-CFP and lipid droplets in CHO cells using fluorescence and CARS microscopy. Perilipin-CFP (cyan), CARS (neutral lipid CH2 vibrations) signal at 2840cm-1 (red). Specimen: H. Nagy, IMB-Graz.
CLICK ON IMAGE TO VIEW THE MOVIE. C. elegans. Lipid droplets imaged at 2840cm-1. Automated 3D tile scan. Frames represent maximum-intensity projections. Leica HCX IRAPO L 25/0.95 W objective. Sampling at 150x150x300 nm.
LASER IS ON!!!!
IMB CARS system based on a Leica SP5 and PicoEMERALD solid-state-laser based light source (APE GmbH; http://www.ape-berlin.de/index_engl.html); HighQ Laser; http://www.highqlaser.at/).
Many thanks to the Leica and APE service team!
Epi-CARS signal of lipid droplets of wildtype yeast cells at 2840 cm-1 (CH2 streching vibration).
Overnight culture in complete medium.
Epi-CARS signal of deutered hexadecanoic acid (C16:0; palmitic acid) crystals at 2105 cm-1 (CD2 streching vibration).
Same crystals, however imaged at 2250 cm-1 (off resonance; maximum NDD voltage).
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