Fu, DanLu, FakeZhang, XuFreudiger, Christian WilhelmPernik, Douglas R.Holtom, GaryXie, Xiaoliang2013-07-162012Fu, Dan, Fa-Ke Lu, Xu Zhang, Christian Freudiger, Douglas R. Pernik, Gary Holtom, and Xiaoliang Sunney Xie. 2012. Quantitative chemical imaging with multiplex stimulated Raman scattering microscopy. Journal of the American Chemical Society 134(8): 3623-3626.0002-7863http://nrs.harvard.edu/urn-3:HUL.InstRepos:10875725Stimulated Raman scattering (SRS) microscopy is a newly developed label-free chemical imaging technique that overcomes the speed limitation of confocal Raman microscopy while avoiding the nonresonant background problem of coherent anti-Stokes Raman scattering (CARS) microscopy. Previous demonstrations have been limited to single Raman band measurements. We present a novel modulation multiplexing approach that allows real-time detection of multiple species using the fast Fourier transform. We demonstrate the quantitative determination of chemical concentrations in a ternary mixture. Furthermore, two imaging applications are pursued: (1) quantitative determination of oil content as well as pigment and protein concentration in microalgae cultures; and (2) 3D high-resolution imaging of blood, lipids, and protein distribution in ex vivo mouse skin tissue. We believe that quantitative multiplex SRS uniquely combines the advantage of fast label-free imaging with the fingerprinting capability of Raman spectroscopy and enables numerous applications in lipid biology as well as biomedical imaging.en-USin-vivoCARS microscopySRS microscopysensitivityJournal Article2013-07-1610.1021/ja210081h