Confocal laser scanning microscopy is one of the most widely used tools for high-resolution imaging of biological cells. However, the imaging resolution of conventional confocal technology is limited by diffraction, and more complex optical principles and expensive optical-mechanical structures are usually required to improve the resolution. This study proposed a deep residual neural network algorithm that can effectively improve the imaging resolution of the confocal microscopy in real time. The reliability and real-time performance of the algorithm were verified through imaging experiments on different biological structures, and an imaging resolution of less than 120 nm was achieved in a more cost-effective manner. This study contributes to the real-time improvement of the imaging resolution of confocal microscopy and expands the application scenarios of confocal microscopy in biological imaging.