We present photophysical properties of functionalized anthradithiophene (ADT) and pentacene (Pn) derivatives, as well as charge and energy transfer properties of donor-acceptor (D/A) pairs of these derivatives. All molecules studied were fluorescent and photostable enough to be imaged on the single-molecule level in a variety of polymeric and in a functionalized benzothiophene (BTBTB) crystalline host using room-temperature widefield epifluorescence microscopy. Flexibility of functionalization of both guest (ADT, Pn) and host (BTBTB or polymer) molecules can be used for systematic studies of nanoscale morphology and photophysics of D/A organic semiconductor bulk heterojunctions, as well as in applications relying on FRET, using single-molecule fluorescence microscopy.