FT-Raman spectroscopy of biological molecules

Abstract

Raman spectroscopy of biological molecules is often very difficult if not impossible due to a large fluorescence background from absorbing species, either from the molecule itself or an impurity. Photobleaching is occasionally successful in photochemically removing fluorescent impurities, but the majority of samples are not responsive to such treatment. Resonance enhancement of an absorbing species allows acquisition of Raman spectra in spite of competing fluorescence. However, the resonance Raman spectrum is characteristic of the chromophore only and little structural information is obtained from the spectrum about other parts of the molecule which are not resonantly enhanced. The newly developed technique of FT-Raman spectroscopy proves to be a solution to both of these problems for biological materials. Excitation with infrared wavelengths prevents electronic absorptions which give rise to fluorescence. In addition, the obtained spectra are completely nonresonant, allowing detection of vibrational modes of all parts of the molecule including the chromophore. We will present nonresonant, fluorescence free spectra of a range of biologically significant molecules including phospholipids and porphyrins.