“…A second important consideration when temperature is varied during the course of an experiment is the fact that the fluorescence itself is affected by temperature, regardless of binding characteristics. In general, quantum yield (Eastman and Rosa, 1968;Song et al, 1975;Cornelissen-Gude and Rettig, 1998;Haynes et al, 1993), and thus fluorescence intensity (Connors et al, 1998;Park, 1996;Law, 1994) both increase as temperature decreases, and, in polar solvents, the effect is even more pronounced (Waris et al, 1988; Bark and Force, 1993). In addition, excited-state lifetimes of fluorescent molecules are strongly affected by temperature, showing, in most cases, longer lifetimes at lower temperatures (Drain et al, 1998;Cornelissen-Gude and Rettig, 1998;Kumke et al, 1997;Young et al, 1997;van den Zegel et al, 1984).…”