The invention discloses a multi-response azacyclo-formaldehyde fluorescent probe molecule with a structure shown as a formula I and a preparation method and application thereof, the molecule realizesselective quantitative recognition of formaldehyde through a fluorescence analysis method, and formaldehyde, copper ions and sulfur ions are tested and evaluated in the fields of automotive trim materials, healthy home furnishing, biological medicines and the like. The multi-response azacyclo-formaldehyde fluorescent probe molecule can be used for fluorescence imaging of biological samples, can also be used as an ultraviolet light absorber to be applied to the technical fields of plastics, coatings, printing ink, display, illumination, glasses, daily-use chemical industry, textiles and the like, and in addition, the multi-response azacyclo-formaldehyde fluorescent probe molecule provided by the invention has relatively strong fluorescence emission performance under both solution and solidconditions; can be used as a fluorescent material in the fields of anti-counterfeiting technologies, light conversion materials, fluorescence labeling, fluorescence imaging and the like, wherein R isH or Cl, and X is CH or N.