From Steady-State to Ultrafast: Resonance Raman Approaches for Biological Samples

Abstract

Vibrational spectroscopy reports on molecular structure with chemical-bond specificity, but in biological systems the vibrational signals of a target chromophore are typically buried under contributions from the surrounding matrix. Resonance Raman (RR) spectroscopy addresses this problem by matching the excitation wavelength to an electronic transition of the chromophore of interest, which increases Raman cross sections by up to six orders of magnitude and restricts the enhanced modes to those coupled to the resonant electronic state. This chapter introduces the physical basis of resonance enhancement and shows how RR isolates chromophore-specific vibrational markers in complex biological systems. We then extend the same principle into the time domain with femtosecond stimulated resonance Raman spectroscopy (FSRRS). In FSRRS, the Raman pump is tuned across the visible range and can be placed in resonance with the transient absorption of a chosen excited-state species, which makes the Raman pump wavelength an additional experimental variable. Sampling this variable across the excited-state absorption manifold provides a selection criterion that separates coexisting transient species sharing a common vibrational window.