NanoLuc substrate 2, a multifunctional dye, serves as an essential tool in various biological experiments. It enables researchers to observe and analyze cell structures, track biomolecules, evaluate cell functions, differentiate cell types, detect biomolecules, examine tissue pathology, and monitor microorganisms. The applications of this dye span from basic scientific research to a broad spectrum of clinical diagnostics. Furthermore, dyes like NanoLuc substrate 2 find extensive use in traditional sectors such as textile dyeing and in newer areas including functional textile processing, food pigments, and dye-sensitized solar cells.