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. 2019 Dec 31;9:20392. doi: 10.1038/s41598-019-56932-8

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Experimental set up for SRS microscope and controls for lipid and protein channels. (A) The Stokes laser is modulated by an electro-optical modulator (EOM) at 20 MHz. Tunable laser is spectrally dispersed through glass rods (tunable) and Stokes laser is dispersed through a grating stretcher (GS). They are the spatially and temporally overlapped at a dichroic mirror (DCM) and directed onto a pair of galvanomirrors (GM). The beams are then sent through a laser scanning microscope with a 25x water immersion objective. The pump beam after the condenser is detected by a photodiode (PD). The signal is processed through lock-in amplifier (LIA). The images are collected using ScanImage on computer processing unit (CPU). (B) SRS spectra for oleic acid and bovine serum albumin (BSA) as controls for lipids and proteins, respectively.