Since the nineteenth century, a great amount of different biological structures and processes have been assessed by fluorescent dyes. Along with the uses of these compounds as vital and histological dyes, some fluorescent dyes have become valuable tools for the study of the pore phenomenon in plasma membranes. Some ion channels capable of forming large conductance channels, such as P2X7, TRPV1, VDAC-1 and the maxi-anion channels transiently alter the plasma membrane permeability, producing pores, which permit the passage of molecules of up to 1,000 Da. In this review, we discuss the uses of the fluorescent dyes chosen in diverse studies of this topic up to now.