ABSTRACT
Total internal reflection fluorescence (TIRF) is an optical technique especially well suited to the study of macromolecule/surface interactions. In particular, TIRF can be used to continuously monitor fluorescent macromolecules either close to (within about 2000 A or less) or directly adsorbed to a solid surface. This chapter provides a brief overview of some selected examples of recent applications of TIRF to the study of protein adsorption to solid surfaces. It begins with a brief overview of theoretical aspects related to TIRF. Factors recognized to be fundamentally relevant to protein/ surface interactions include adsorption, desorption, and diffusion kinetics, chemical equilibrium between surface-adsorbed proteins and free solution proteins, effects of the hydrodynamic environment on the adsorption process, and the conformation of the adsorbed protein layer. TIRF studies of protein adsorption can be divided into two categories: the quantitative investigation of kinetic, diffusion, and equilibrium characteristics; and the investigation of adsorbed protein conformation.
