Molecular techniques are transforming our understanding of cellular function and disease. However, accurate molecular analysis methods will be limited if the input DNA, RNA, or protein is not derived from pure population of cells or is contaminated by the wrong cells. A novel Ultrasonic Vibration Microdissection (UVM) method was proposed to procure pure population of targeted cells from tissue sections for subsequent analysis. The principle of the Ultrasonic Vibration Cutting is analyzed, and a novel microknife is designed. A multilayer piezoelectric actuator is used to actuate a sharp needle vibrating with high frequency and low amplitude (Approx. 16–50 kHz, and 0-3μm) to cut the tissue. Contrast experiment was done to test the feasibility of UVM method. Experimental results show that the embedded tissue can be quickly and precisely cut with the ultrasonic vibration micro-dissection method.