Lignin is an abundant biomass material in the natural world, its application value and application scope ought to be enlarged. In this work, dealkalized lignin with good water-solubility was used to combine with water-insoluble boron-dipyrromethene (BODIPY) fluorophore to obtain readily water-soluble macromolecular probes. Firstly, a small molecular fluorescent BODIPY probe with two amino groups (-NH2) and another small molecular fluorescent probe with two chlorine groups (-Cl) were synthesized. Secondly, the small-molecular fluorescent probe with two -NH2 groups reacted with carbonyl groups in the lignin by Schiff base formation reaction. Meanwhile, the small-molecular fluorescent probe with two -Cl groups reacted with hydroxyl groups in the lignin by Williamson ether formation reaction. Eventually, two lignin-based fluorescent probes were obtained. The molecular weights of the two probes reached 22.4 kDa and 20.1 kDa. Moreover, these two fluorescent probes were readily soluble in H2O and DMF. Both of the fluorescent macromolecules have good recognition ability for Fe3+, with detection limits of 2.1 × 10−7 mol/L and 1.1 × 10−7 mol/L respectively. Fe3+ interacts with the fluorescent probe, and the imine bond (-CN-) reacts with Fe3+ to cause fluorescence response. The photo-induced electron transfer effect is enhanced, and the photo-induced electron transfer (PET) mechanism causes the fluorescent quenching. This work expands the application scope of lignin as a biomass material and lays a foundation for the development of lignin materials.