Imaging of hepatocellular nitric oxide (NO) in vivo is of great importance for understanding its roles in liver diseases, but such an imaging approach is still lacking. Herein, a hepatocyte-targeting fluorescent NO sensor has been fabricated. This sensor exhibits good water solubility, excellent selectivity, and high sensitivity (∼1.62 nM). The cells and in vivo imaging experiments reveal that Gal-RhB facilitates the visualization of hepatocellular NO in both HepG2 cells and zebrafish. These results demonstrate that Gal-RhB may be a promising tool for studying the function of NO in NO-associated liver diseases.