Multiphoton Microscopy for Visualizing Lipids in Tissue. Author: Lee M1, Serrels A2 Affiliation: <sup>1</sup>Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, EH4 2XU, Scotland, UK. martin.lee@ed.ac.uk. <sup>2</sup>Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, EH4 2XU, Scotland, UK. Conference/Journal: Methods Mol Biol. Date published: 2016 Other: Volume ID: 1467 , Pages: 105-18 , Special Notes: doi: 10.1007/978-1-4939-4023-3_9. , Word Count: 132 Visualizing the appearance of fat droplets and adipocytes in tissue can be realized using a label-free imaging method known as coherent anti-Stokes Raman spectroscopy (CARS). CARS is a nonlinear optical technique that allows label-free imaging of a material with contrast based on the same vibrational signatures of molecules found in Raman spectroscopy. CARS can be combined with other single and multiphoton imaging modes such as second harmonic generation and two-photon fluorescence to image a broad variety of biological structures.Here we describe the construction of a multiphoton microscope that will enable the study of both fluorescently labeled and unlabeled tissue. This has been used to monitor the contribution of Wt1 expressing cells towards the visceral fat depots during gestation. KEYWORDS: CARS; Lipids; Microscopy; Multiphoton; Raman; SHG PMID: 27417963 DOI: 10.1007/978-1-4939-4023-3_9