Introduction

Abstract

Because of their extraordinary sensitivity and speed, optical spectroscopic techniques are well suited for addressing a broad range of questions in molecular and cellular biophysics. Photomultipliers sensitive enough to detect a single photon make it possible to measure the fluorescence from individual molecules, and lasers providing light pulses with widths of less than 10−14 s can be used to probe molecular behavior on the time scale of nuclear motions. Spectroscopic properties such as absorbance, fluorescence and linear and circular dichroism can report on the identities, concentrations, energies, conformations and dynamics of molecules and can be sensitive to small changes in molecular structure or surroundings. Resonance energy transfer provides a way to probe intermolecular distances. Because they usually are not destructive, spectrophotometric techniques can be used with samples that must be recovered after an experiment. They also can provide analytical methods that avoid the need for radioisotopes or hazardous reagents. When combined with genetic engineering and microscopy, they provide windows to the locations, dynamics and turnover of particular molecules in living cells.