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Electrostatic interactions impact every aspect of the structure and function of proteins, nucleic acids, and membranes.The transition states for many enzyme-catalyzed reactions involve a change in the distribution of charge relative to the starting material and/or products, and the selective stabilization of charge-separated transition states may be essential for catalysis.The magnitudes of the electric fields in proteins and the variations in these fields at different sites are predicted to be enormous, but it is a challenge to obtain quantitative experimental information on these fields.We have developed vibrational Stark effect (VSE) spectroscopy to probe electrostatics and dynamics in organized systems, in particular in proteins where they can report on functionally important electric fields.