Abstract: The realization of high sensitivity detection of pesticide residues has important significance and research value for understanding environmental problems and protecting human life safety. Nano-fluorescent probes can be used for rapid in-situ detection of pesticide residues, but there are many problems, such as high cost, high consumption, long preparation period of fluorescent probes, harm of toxic and harmful substances to laboratory personnel and environmental pollution. Through the transformation of scientific research achievements, the virtual simulation platform for the detection of pesticide residual pollutants by nano-fluorescence probe was designed and developed based on 3D virtual technology. The experimental platform was equipped with 7 modules, including preparation of nano-fluorescent probe, preparation of standard sample, pre-treatment of fruit and vegetable sample, pre-treatment of agricultural wastewater sample, preparation of detection sample, analysis of detection performance of nano-fluorescent probe, and mechanism of interaction between nano-fluorescent probe and detection object. Through the virtual simulation experiment platform, students can master the synthesis of materials, the operation of instruments, the observation of experimental phenomena, and the processing and analysis of experiment data. By means of the teaching platform, students can use virtual experiments to cultivate their comprehensive knowledge application ability, autonomous learning ability, problem-solving ability and hands-on practice ability, and enhance students’ awareness of scientific research innovation.