拉曼 Raman spectroscopy.ppt

UV and Resonance Raman Spectroscopy UV lasers allow for better Raman performance, because of the 1/?4 dependence of scattering, but fluorescence is a problem With lasers in the 245-266 nm region, the Raman spectrum can be “fit” in the region above the laser but below the normal Stokes-shifted fluorescence spectrum UV and Resonance Raman Spectroscopy Resonance Raman scattering excites an electronic transition (e.g. using a UV laser in the 240-270 nm range) Transitions can achieve 1000x increase in signal Raman Resonance Raman Surface Enhanced Raman Spectroscopy (SERS) SERS is a form of Raman spectroscopy that involves a molecule adsorbed to the surface of a nanostructured metal surface which can support local surface plasmon resonance (LSPR) excitations The Raman scattering intensity depends on the product of the polarizability of the molecule and the intensity of the incident beam; the LSPR amplifies the beam intensity when the beam is in resonance with plasmon energy levels – leads to signal enhancements of 106 Single-molecule detection with SERS has been demonstrated R. A. Halverson, P. J. Vikesland, Environ. Sci. Technol. 2010, 44, 7749–7755, /10.1021/es101228z Coherent Anti-Stokes Raman Spectroscopy (CARS) In CARS, the sample is excited by a probe beam with frequency ?pump, a Stokes beam (?Stokes) and a probe beam (?probe) CARS uses tightly focused beams delivered via a microscope to achieve a phase matching condition necessary for the coherent process Scanning a sample using a given vibrational resonance frequency can be used to determine the spatial distribution a Raman-active vibrational transitions at this frequency CARS Applications CARS is commonly used to perform rapid chemical imaging of biological materials for these components DNA (phosphate stretching vibration) Protein (amide I stretch) Water (OH stretch) Lipids (CH vibrations – stretching, bending, etc…) Video-rate imaging of cells has been demonstrated C. L. Evans, X. S. Xie, Annu. Rev. Anal. Chem