The imaging capability(sensitivity, signal-to-noise ratio, imaging speed and spatial resolution) of a domestic-made high-resolution Hadamrd transform(HT) microscopic fluorescence imaging system was discussed. The results show that the system can be applied for high-resolution single-cell imaging sensitively in a short time, the cell image provided by the system has a high-resolution up to 511×512 pixel. The results also shows that this system has a more excellent resolving power than conventional optical microscope, under a 100× objective(oil lens) the spatial resolution up to 0.24 μm per pixel in X direction was obtained, which is close to the resolution limit of conventional optical microscope. The system was used for multi-parameter evaluation of breast tumor malignancy and malignant grade for a single tumor cell based on nuclei DNA ploidy measurements, and satisfactory results were obtained.