Microinjection is a widely used technique for introducing exogenous materials into cells. Many applications of microinjection, such as gene editing and drug testing, rely on the accurate control of the deposition volume. However, the deposition volume in microinjection is presently calibrated in an open medium without considering the cell inner pressure effect, which we experimentally show in this paper that it can induce an error as large as 30% between the actual deposition volume and the set volume. In this work, the relationship between the cell inner pressure and the deposition volume was analytically modeled and experimentally validated. On the basis of the developed model, the cell inner pressure of a given cell type can be well estimated from the injection pressure and the resulting deposition volume. The quantitated cell inner pressure is then used to reduce the error between the set volume and the actual deposition volume. Experiments conducted on human bladder cancer cells (T24 and RT4) showed that T24 cells have a higher inner pressure than RT4 cells (405 ± 45 Pa vs 341 ± 34 Pa), and after compensating for the cell inner pressure, the error between the intended set volume and the actual deposition volume into a cell became less than 3%.