Provided is an image deblurring algorithm based on a sparse positive source separation model. The image deblurring algorithm is used for performing processing on a blurry image collected by an optical microscopic imaging system and generated due to diffraction effect and optical deviation; under a condition of single light sensitive imaging and of not increasing an external imaging device, a spatial resolution of the optical microscopic system may be improved to a nanoscale. In the disclosure, a blurring process of microscopic imaging is expressed as a linear combination of a Point Spread Function of the imaging system; by embedding the process into a positive separation optimized frame, adding a sparsity constraint and solving to remove blurring, the high-resolution microscopic imaging is implemented.