The invention discloses a dustless filter film storage box for dust sampling and an application method of the dustless filter film storage box, and belongs to the technical field of sampling auxiliary devices. The dustless filter film storage box comprises a filter film box barrel, a storage barrel, a supply barrel, a bearing sheet, a spring, a filter film box cover, a spiral center rod, a non-closed spiral slope face plate, a double-tweezers-sheet clamping device, a thin metal sheet, a kneading key cap, a sliding arc groove, a bottom plate, a filter film box top cover and rotation cover sheets. Through the design, the filter film storage box can provide enough sampling filter film reserve parts for storing filter films for field sampling, and meanwhile the occupied space is greatly saved in size. Through the design of the supply barrel, automatic popup can be conducted for filter film supplementing along with reduction of the number of filter film sheets in the filter film usage process. A clamping tool is internalized into the double-tweezers-sheet clamping device, and an operation method is simplified. Due to the double-layer rotation cover sheet design on the filter film box top cover, simplification is achieved, operation is easy, and the operation error rate can be reduced; and a more reliable guarantee is provided for preventing to-be-used filter films from being polluted by the outside through various internalized operation components.