Institute for Photonics and Nanotechnologies (IFN-CNR) 1 article published in JoVE Engineering Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy Alejandro De la Cadena1, Federico Vernuccio1, Benedetta Talone1, Arianna Bresci1, Chiara Ceconello1, Subir Das1, Renzo Vanna2, Giulio Cerullo1,2, Dario Polli1,2 1Dipartimento di Fisica, Politecnico di Milano, 2Institute for Photonics and Nanotechnologies (IFN-CNR) We present a protocol to acquire chemical images with broadband stimulated Raman scattering (SRS) microscopy. Based on an SRS microscope that operates with differential multichannel-lock-in detection, the protocol describes the sample preparation, adjustment of the SRS apparatus, and chemometrics to disentangle different constituents of chemically heterogeneous samples.