Fast and long term lipid droplet tracking with CARS microscopy

Photobleaching of organic fluorophores commonly used in fluorescence microscopy puts a limit to the number of images which can be acquired. Label-free imaging techniques therefore offer advantages both for rapid image acquisition and for long-term observations. CARS microscopy is a label-free imaging technique offering molecule specific contrast. Here we demonstrate that CARS microscopy allows video-rate tracking of intracellular transport of lipid droplets, but also continuous long-term observation of cells over several hours.

[1]  X. Xie,et al.  Nonperturbative chemical imaging of organelle transport in living cells with coherent anti-stokes Raman scattering microscopy. , 2006, Biophysical journal.

[2]  L M Loew,et al.  High-resolution nonlinear optical imaging of live cells by second harmonic generation. , 1999, Biophysical journal.

[3]  H. Jäckle,et al.  Lipid droplets: A dynamic organelle moves into focus , 2010, FEBS letters.

[4]  Yohanns Bellaiche,et al.  Tracking individual kinesin motors in living cells using single quantum-dot imaging. , 2006, Nano letters.

[5]  Robert G. Parton,et al.  Opinion: Lipid droplets: a unified view of a dynamic organelle , 2006, Nature Reviews Molecular Cell Biology.

[6]  E. Wieschaus,et al.  Dynein-mediated cargo transport in vivo. A switch controls travel distance. , 2000 .

[7]  S. Gambhir,et al.  Quantum Dots for Live Cells, in Vivo Imaging, and Diagnostics , 2005, Science.

[8]  Steven P. Gross,et al.  Consequences of Motor Copy Number on the Intracellular Transport of Kinesin-1-Driven Lipid Droplets , 2008, Cell.

[9]  J. Füllekrug,et al.  Lipid droplets lighting up: Insights from live microscopy , 2010, FEBS letters.

[10]  W. R. Wiley,et al.  Three-Dimensional Vibrational Imaging by Coherent Anti-Stokes Raman Scattering , 1999 .

[11]  T. Kreis,et al.  Translocation and clustering of endosomes and lysosomes depends on microtubules , 1987, The Journal of cell biology.

[12]  D Yelin,et al.  Laser scanning third-harmonic-generation microscopy in biology. , 1999, Optics express.

[13]  M. Welte.  Fat on the move: intracellular motion of lipid droplets. , 2009, Biochemical Society transactions.

[14]  Kevin Braeckmans,et al.  The transport of nanosized gene carriers unraveled by live-cell imaging. , 2006, Angewandte Chemie.

[15]  Petros Koumoutsakos,et al.  A novel supervised trajectory segmentation algorithm identifies distinct types of human adenovirus motion in host cells. , 2007, Journal of structural biology.

[16]  Rodney K. Lyn,et al.  Dynamics of lipid droplets induced by the hepatitis C virus core protein. , 2010, Biochemical and biophysical research communications.

[17]  Michael J Rust,et al.  Visualizing infection of individual influenza viruses , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[18]  A. Fabre,et al.  Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy , 2005, Nature Methods.

[19]  M. Orrit,et al.  Absorption and scattering microscopy of single metal nanoparticles. , 2006, Physical chemistry chemical physics : PCCP.

[20]  J. Peychl,et al.  Live Cell Multicolor Imaging of Lipid Droplets with a New Dye, LD540 , 2009, Traffic.

[21]  D. Débarre,et al.  Quantitative characterization of biological liquids for third-harmonic generation microscopy. , 2007, Biophysical journal.

[22]  Michitaka Suzuki,et al.  A pitfall in using BODIPY dyes to label lipid droplets for fluorescence microscopy , 2010, Histochemistry and Cell Biology.

[23]  Andreas Zumbusch,et al.  Coherent anti-Stokes Raman scattering microscopy , 1999 .