Fluorescence imaging of subcellular structures is crucial for understanding cellular functions and disease mechanisms. However, existing fluorescent dyes are plagued by inadequate biocompatibility and insufficient stability, which impede the advancement of high-resolution, long-term, and live-cell imaging. In this work, we innovatively synthesized a novel nickel nanocluster encapsulated with ficin (Ficin-Ni NCs) through a straightforward biomineralization approach as a fluorescent probe for imaging the nucleus. The Ficin-Ni NCs with intense green fluorescence exhibited a particle size of approximately 1.56 nm, significant Stokes shift (110 nm), and remarkable stability (98 % fluorescence intensity after 14 days). Meanwhile, the Ni NCs demonstrated excellent biocompatibility, maintaining cell viability above 90 % for both Raw264.7 and HEK-293T cells at a concentration of 60 μg/mL. Significantly, DNA gel electrophoresis experiments, using ethidium bromide (EB) as a reference, confirmed the ability of the NCs to bind DNA. Furthermore, the as-prepared Ficin-Ni NCs could achieve precise imaging of cell nuclei consistent with 4',6-diamidino-2-phenylindole (DAPI) within 15 min in a variety of animal cells, including HEK-293T cells, Raw264.7, bullfrog erythrocytes, and human oral epithelial cells. Simultaneously, Ni NCs also demonstrated remarkable potential for visualizing the nuclear structures in plant cells, such as onion and cinnamon epidermal cells. Therefore, this innovative fluorescent probe, initially prepared via biomineralization, emerges as a promising alternative to traditional EB or DAPI dyes, paving the way for rapid response, effectiveness, and reusability in cellular imaging.