To reveal the penetration mechanism and present the penetration characteristics of high-speed micro-jet with injection volume larger than 0.3mL into soft tissue, the present study conducted experimental research on high-speed water-jet penetration into ballistic gelatin. The free jet dynamics of an air-powered needle-free injector that can emit up to 1.27mL of liquid at once and the penetration dynamics were visualized to reveal the details of the penetration process. In the early unstable stage, the jet is emitted in the form of pulses, and the first jet pulse can rapidly generate an initial slender channel in gelatin in a very short time. In the subsequent stable stage, energy input produces dispersion and further increases the penetration depth slowly. Changing the driving pressure by the power source mainly changes the penetration depth increment by dispersion; while changing the nozzle diameter mainly affects the penetration depth in the initial stage. The central position of the dispersion area in the injection direction was firstly defined in the present work and it was found that an approximate linear relationship between this position and the maximum penetration depth exits for different nozzle diameters and driving pressures when injecting the same liquid dose. These research results can provide a basis for a thorough understanding of the penetration characteristics of high-speed micro-jet with injection volume larger than 0.3mL into soft tissue, as well as the design and operation of the air-powered needle-free injector.