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Shedding new light on lipid functions with CARS and SRS microscopy
Authors:Yong Yu  Prasanna V RamachandranMeng C Wang
Institution:Huffington Center on Aging, Baylor College of Medicine, Houston, TX, USA; Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA
Abstract:Modern optical microscopy has granted biomedical scientists unprecedented access to the inner workings of a cell, and revolutionized our understanding of the molecular mechanisms underlying physiological and disease states. In spite of these advances, however, visualization of certain classes of molecules (e.g. lipids) at the sub-cellular level has remained elusive. Recently developed chemical imaging modalities – Coherent Anti-Stokes Raman Scattering (CARS) microscopy and Stimulated Raman Scattering (SRS) microscopy – have helped bridge this gap. By selectively imaging the vibration of a specific chemical group, these non-invasive techniques allow high-resolution imaging of individual molecules in vivo, and circumvent the need for potentially perturbative extrinsic labels. These tools have already been applied to the study of fat metabolism, helping uncover novel regulators of lipid storage. Here we review the underlying principle of CARS and SRS microscopy, and discuss the advantages and caveats of each technique. We also review recent applications of these tools in the study of lipids as well as other biomolecules, and conclude with a brief guide for interested researchers to build and use CARS/SRS systems for their own research. This article is part of a Special Issue entitled Tools to study lipid functions.
Keywords:CARS  Coherent Anti-Stokes Raman Scattering  SRS  Stimulated Raman Scattering  CRS  Coherent Raman Scattering  PMT  photomultiplier tube  OPO  optical parametric oscillator  SRL  Stimulated Raman Loss  SRG  Stimulated Raman Gain  TAG  triacylglycerol  CGI-58  comparative gene identification 58  3D  three-dimensional  DMSO  dimethyl sulfoxide  ps  picosecond  IR  infrared  DC  direct current  N  A    numerical aperture  A/D  analog-to-digital
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