Intracytoplasmic sperm injection (ICSI), as the savior of male infertility, has been applied to artificial assisted reproduction (IVF). The oocytes are inevitably ruptured by the microinjector in the process of microoperatin. To study the effects of mechanical damage from microopetations on embryo development, we analyzed 60 the spent culture media of porcine parthenogenetic embryos experienced different types of microopetation using Raman spectroscopy. The results showed that Group II with the puncture of egg zona and plasma membrane had the highest cleavage rate and blastocyst rate of porcine parthenogenetic embryos, which was significantly higher than Group I with intact oocytes (P<0.05) and Group III with the puncture of zona and plasma membrane and aspiration egg cytoplasm (P<0.05) on the cleavage rate, and was extremely significantly higher than Group III on the blastocyst rate (P<0.001). Raman spectroscopy of the spent culture media of these parthenogenetic embryos revealed that there were obvious reduced in the intensity of Raman characteristic peaks at 810 cm-1, 1371 cm-1 and 1563 cm-1 in the microoperation groups (Group II and Group III). These changes of Raman characteristic peaks suggested that ICSI had influenced the metabolism of embryonic membrane lipids, membrane proteins and nucleic acids. The PCA analysis also exhibited that these three groups were well distributed in different areas. In conclusion, the microoperation of ICSI caused some changes in the metabolism of embryos. Raman spectroscopy is a valuable technology for assessing embryonic metabolism.