Using maleimidoethylmonoamide cysteine (Fmoc)(StBu) (1) as a medium, cysteine residues on proteins of chicken eggshell membrane (ESM) were successfully converted into N-terminal cysteines. After a biocompatible condensation reaction between the N-terminal cysteine and fluorescent probe 2-cyanobenzothiazole-Gly-Gly-Gly-fluorescein isothiocyanate (2), a new fluorogenic structure luciferin-Gly-Gly-Gly-FITC (3) was obtained, which exhibits a 2-fold fluorescence emission increase compared to that of 2. Thus, a new method for effectively labeling cysteine residues on ESMs was developed. Enhanced fluorescence images of ESMs were directly observed under a microscope and a small animal imaging machine.