Lithium-ion batteries (LIBs) have become a main energy storage device in various applications, such as portable appliances, renewable energy facilities, and electric vehicles. However, the poor thermal stability of LIBs may cause explosion or fire. The thermal runaway is the result of a failure of the separator inside LIB. Damages like tearing, piercing, and collapsing of the separator were simulated in a mechanical, an electrical, and a thermal way, and small discharge pulses of a few mV were detected at the time of separator damages. From the experimental results, this paper provided a method that can identify the separator failure before thermal runaway in the aspect of a potential explosion and fire prevention measures.