A newly synthesized imine-based receptor (L) showed remarkable specificity toward the Hg2+ ion in aqueous media over other metal ions. Coordination of L to Hg2+ induces a turn-on fluorescence response. This was explained based on the restricted imine isomerization along with PET on coordination to Hg2+. X-ray structural evidence tends to favor a C鈥揅 bond rotation rather than C鈺怤 isomerization for adopting a favorable conformation in L for coordination to Hg2+. This reagent could be used for imaging the accumulation of Hg2+ ions in HeLa cells.