Abstract: In order to obtain the nucellus of Paeonia suffruticosa, the non-enzymatical isolating technique was studied on the basis of structural observation of their ovules. The ovules, fresh and fixed by alcohol solution of P. suffruticosa were used to observe its structure characteristics with the whole clearing technique by 84 disinfectants and the slicing technique under a fluorescence microscope, and their fresh ovules were still used to isolate its nucellus and other structures with gum block under a stereomicroscope. Results identified the outer integument of P. suffruticosa ovules was extremely thick and easy to be peeled off. The outer layer of the inner integument of P. suffruticosa could be separated and isolated easily from the integument tapetum. In whole ovule only the integument tapetum could emit clear fluorescence. After the outer integument, the outer layer of the inner integument and the integument tapetum were peeled off successively with gum block under a stereomicroscope, the nucellus with original form could be isolated.