【Objective】 This research aims to explore the effects of different storage time on the separation effects of soil animals under the condition of soil low temperature storage, so as to provide a reference for a large number of collection and separation of soil animals.【Method】 On October 6, 2018, the natural Mongolian oak (Quercus mongolica) forest in Songhua Lake Nature Reserve, Jilin City, the soil at different depths was drilled by the five point sampling method, brought back to the laboratory and stored in the indoor 4 ℃ refrigerator. The Berlese-Tullgren method was used to separate the soil animal samples of Oribatida and Collembola on the same day, 7, 14, 21 and 28 days later. The species and genera of Oribatida and Collembola were determined according to the morphological characteristics under the microscope.【Result】 With the increase of preservation time, the individual number and the number of species of soil animals of Oribatida and Collembola showed a downward trend; Under the same preservation time and different depths, the species of Oribatida and Collembola had significant differences with other depths (P<0.05), and showed a downward trend. The number of species of Oribatida and Collembolan collected in litter layer was significantly different from that in 5, 10 and 15 cm soil layers (P<0.05), and also showed a downward trend.【Conclusion】 Increasing the soil preservation time under cold storage will reduce the separation efficiency of medium-sized soil animals. The separation effect of Oribatida is the best on the day of soil collection, and the isolation of Collembola was not affected by soil storage time. Oribatida and Collembola animals were mainly collected in the litter layer. The soil depth increased while the soil animals decreased.