In the anticancer research with traditional Chinese medicine, many medicinally effective components can only dissolve in higher polar organic solvents, such as ethanol, dimethyl sulfoxide (DMSO) etc. However, organic solvents may directly interfere with the accuracy of therapeutic efficacy evaluation. Therefore the study on effects of organic solvents with different concentrations on Hela cells is of great significance. The digital holography is a non-destructive and non-contact method to image the transparent sample without staining and with the high precision and high resolution. In this paper, the digital holography is proposed to replace the methyl-thiazol-tetrazolium (MTT) or the Giemsa dye method. Based on the pre-magnification off-axis Fresnel digital holographic theory, an inverted microscopy system is built to obtain the phase-contrast images of the Hela cells, which are added different concentrations of organic solvents. Compared to the control group, there is significantly differences with the shapes of Hela cells with different organic solvents. The size of cell with ethanol 25% is no significantly difference with the control group. But the sizes of cells in the solutions with ethanol 12.5% and 50% are smaller than the control group. Next, the sizes of cells in the solutions with DMSO 12.5%, 25% and 50% are great smaller, compared with the control group. The results show that the digital holography has high practical value in detecting the changes in the shape of cells and is helpful in the choice of organic solvents for further apoptosis study.