Electrophoresis in its various forms has had an enormous impact on biochemistry. The primary use has been the quantitative determination of electrophoretically distinct species in a mixture. A second, less common, type of application is the interpretation of the elctrophoretic mobility in terms of the surface charge of the species. The latter application has been hindered both by experimental difficulties of measuring the electrophoretic mobility and by the theoretical difficulties of relating the electrophoretic mobility to the number of electronic charges on the surface of the particle. Classical electrophoresis techniques measure the distance travelled per unit time by a charged particle in an electric field. Since electrophoretic velocities are small, the time required for transit of a measurable distance can be inconveniently long. However, if the velocity could be measured directly, then the electrophoresis experiment could be accomplished without waiting for macroscopic migration, and electrophoretically distinct species could be determined without requiring theirphysicalseparation.Velocitymeasurements of macroscopic objects are commonplace. Everyone is familiar with the use of radar for determining the velocity of an object by measuring the Doppler shift of the microwave radiation reflected from the object. The development of the optical laser has made it possible to use the Doppler principle to determine the velocities of particles of all sizes by measuring the frequency shift of the laser light which has been scattered from them. This new technique, usually called laser Doppler velocimetry, has already been applied to several interesting problems in biology and medicine [ 11. Determination ofelectrophoretic velocities by laser Doppler velocimetry is known as electrophoretic light scattering. The theory of electrophoretic light scattering and the first successful experiments were reported in 197 1 by Ware and Flygare [2]. Since that time the technique has been refined and adapted by several groups, and over thirty publications have appeared, including applications to proteins, nucleic acids, viruses, bacteria, blood cells and synthetic polyelectrolytes. The information obtained in these experiments always includes the electrophoretic mobility of the species being studied. In heterogeneous samples, the mobility of several species can be measured simultaneously, since each contributes a spectral peak corresponding to the Doppler shift of its characteristic velocity. In many cases the width of the peak in the electrophoretic light scattering spectrum is attributable solely to diffusion and can be used to determine the diffusion coefficient of that particular species. The linewidth can therefore be used as an indicator of macromolecular conformation or the state of aggregation or fusion of the observed species. In rhe case of very large particles such as blood cells, the diffusion broadening is negligible, and the electrophoretic light scattering spectrum is a simple histogram representing the electrophoretic distribution of the sample. spectrum analyzer or a Fourier transform minicomputer, and miscellaneous optical and electronic components. In addition, a special electrophoretic light-scattering chamber must be fabricated. Once a satisfactory apparatus has been assembled, experiments can be performed rapidly and efficiently. The complete electrophoretic spectrum of a sample can be measured in as little time as a few seconds and rarely takes longer than a few minutes. Signal-to-noise ratios of 100 or greater are common, and the precision of an electrophoretic mobility determination is typically between 1% and 5%. The technique is ideal for repeated analysis of a sample which is changing in time or for routine analysis of a large number of samples. Adaptation for on-line repeated analyses would be a straightforward modification of existing equipment. Electrophoreticlight scatteringmeasurements are performed on solutions or suspensions of charged particles. Attempts to utilize this technique to detect electrophoretic motion on supporting media such as paper or gels have not been successful Therefore, the high resolution among species obtainable by electrophoresis on supporting media is not matched by electrophoretic light scattering. For proteins and other macromolecules of molecular weight less than one million, the principal uses of this new technique will probably be the measurement of solution electrophoretic mobilities and the study of interacting systems.