title: Surface plasmon coupled emission in micrometer-scale cells: a leap from interface to bulk targets.
abstract: Surface plasmon coupled emission (SPCE) technique has attracted increasing attention in biomolecular interaction analysis and cell imaging because of its high sensitivity, low detection volume and low fluorescence background. Typically, the working range of SPCE is limited at nanometers to an interface. For micrometer-scale samples, new SPCE properties are expected because of complex coupling modes. In this work, cells with different subregions labeled were studied using a SPCE spectroscopy system. Angular and p-polarized emission was observed for cell membrane, cytoplasm, and nucleus labeled with DiI, Nile Red, and propidium iodide, respectively. The SPCE signals were always partially p-polarized, and the maximum emission angle did not shift, regardless of variations in emission wavelength, fluorophore distribution and stained layer thickness. Additionally, increased polarization and a broader angle distribution were also observed with an increase in sample thickness. We also investigated the impact of metallic substrates on the SPCE properties of cells. Compared with Au and Ni substrates, Al substrates presented better polarization and angle distribution. Moreover, the real-time detection of the cell labeling process was achieved by monitoring SPCE intensity. These findings expand SPCE from a surface technique to a 3D method for investigating bulk targets beyond the nanoscale interfaces, providing a basis to apply this technique to study cell membrane fluidity and biomolecule interactions inside the cell and to distinguish between cell subregions.
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University , Xiamen 361005, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.forename: Shuo-Hui
authors.initials: SH
authors.lastname: Cao
authors.forename: Wei-Peng
authors.initials: WP
authors.lastname: Cai
authors.forename: Xiao-Qing
authors.initials: XQ
authors.lastname: Liu
authors.forename: Yu-Hua
authors.initials: YH
authors.lastname: Weng
authors.forename: Kai-Xin
authors.initials: KX
authors.lastname: Xie
authors.forename: Si-Xin
authors.initials: SX
authors.lastname: Huo
authors.forename: Yao-Qun
authors.initials: YQ
authors.lastname: Li
mesh_terms: D002232:Carbocyanines; D003198:Computer Simulation; D005240:Feasibility Studies; D006046:Gold; D006367:HeLa Cells / Q000737:chemistry* / Q000166:cytology*; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D018613:Microscopy, Confocal; D008954:Models, Biological; D009532:Nickel; D010078:Oxazines; D019904:Polymethyl Methacrylate; D011419:Propidium; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: C582073:1,1'-dioleyl-3,3,3',3'-tetramethylindocarbocyanine; D002232:Carbocyanines; D010078:Oxazines; D011419:Propidium; D006046:Gold; D009532:Nickel; D019904:Polymethyl Methacrylate; C044808:nile red