In situ Plasmon-Enhanced CARS and TPEF for Gram staining identification of non-fluorescent bacteria.

Abstract

In this work, we report in situ nonlinear microscopic images on plasmon-enhanced coherent anti-Stokes Raman scattering (CARS) and plasmon-Induced two-photon excited fluorescence （TPEF)of non-fluorescent microorganism. Our unique synthesized Au@Ag nanorods provide with two distinct surface-plasmon resonance (SPR) at 400 and 800\xa0nm, respectively, which can efficiently induce linear fluorescence signals of E. coli but also enhance the nonlinear optical spectroscopy signals of TPEF, and coherent anti-Stokes Raman scattering (CARS) imaging of E. coli and S. aureus. Furthermore, calculations with complete active space self-consistent field (CASSCF) reveals the hot electrons of SPs can efficiently induce the biological fluorescence of non-fluorescent flavin nucleotides on the surface of E. coli. This novel mechanism is expected to guide the development and application of new microbial detection reagents. Gram-negative and Gram-positive bacteria can be well distinguished by nonlinear microscopic imaging of the CARS signal at 1589\xa0cm-1. Benefit by the strong penetrability of non-linear optical signals, it is expected to realize in situ real-time detection and classification of pathogenic microbial infections in vivo.