Zong, ChengPremasiri, RanjithLin, HaonanHuang, YiminZhang, ChiYang, ChenRen, BinZiegler, Lawrence D.Cheng, Ji-Xin2020-04-172020-04-172019-11-21Cheng Zong, Ranjith Premasiri, Haonan Lin, Yimin Huang, Chi Zhang, Chen Yang, Bin Ren, Lawrence D Ziegler, Ji-Xin Cheng. 2019. "Plasmon-enhanced stimulated Raman scattering microscopy with single-molecule detection sensitivity.." Nat Commun, Volume 10, Issue 1, pp. 5318. https://doi.org/10.1038/s41467-019-13230-12041-1723https://hdl.handle.net/2144/40234Stimulated Raman scattering (SRS) microscopy allows for high-speed label-free chemical imaging of biomedical systems. The imaging sensitivity of SRS microscopy is limited to ~10 mM for endogenous biomolecules. Electronic pre-resonant SRS allows detection of sub-micromolar chromophores. However, label-free SRS detection of single biomolecules having extremely small Raman cross-sections (~10-30 cm2 sr-1) remains unreachable. Here, we demonstrate plasmon-enhanced stimulated Raman scattering (PESRS) microscopy with single-molecule detection sensitivity. Incorporating pico-Joule laser excitation, background subtraction, and a denoising algorithm, we obtain robust single-pixel SRS spectra exhibiting single-molecule events, verified by using two isotopologues of adenine and further confirmed by digital blinking and bleaching in the temporal domain. To demonstrate the capability of PESRS for biological applications, we utilize PESRS to map adenine released from bacteria due to starvation stress. PESRS microscopy holds the promise for ultrasensitive detection and rapid mapping of molecular events in chemical and biomedical systems.p. 5318en-US© The Author(s) 2019. This article is licensed under a Creative Commons Attri-bution 4.0 International License, which permits use, sharing, adaptation,distribution and reproduction in any medium or format, as long as you give appropriatecredit to the original author(s) and the source, provide a link to the Creative Commonslicense, and indicate if changes were made. The images or other third party material in thisarticle are included in the article’s Creative Commons license, unless indicated otherwise ina credit line to the material. If material is not included in the article’sCreativeCommonslicense and your intended use is not permitted by statutory regulation or exceeds thepermitted use, you will need to obtain permission directly from the copyright holder.http://creativecommons.org/licenses/by/4.0/AdenineElectronsLasersNitrogen isotopesNonlinear optical microscopyReproducibility of resultsSingle molecule imagingSpectrum analysis, RamanStaphylococcus aureusStress, physiologicalArticle10.1038/s41467-019-13230-10000-0001-9454-847X (Yang, Chen)0000-0002-5607-6683 (Cheng, Ji-Xin)494079