The confocal imaging has become one of the most widely applied microscopic techniques in various fields, such as biotechnology, automation engineering, optical engineering, solid-state physics, metallurgy, integrated circuit inspection, etc. Confocal laser scanning microscopy (CLSM) is primarily based on the use of apertures in the detection path to provide the acquired three-dimensional images with satisfactory contrast and resolution. The major objective of this paper is to analyze the imaging performance of the confocal microscopes with varying opto-mechanical conditions. In this paper, the working principles of the one- and two-dimensional scanning mechanisms in the microscopic systems are first reviewed and verified by opto-mechanical simulations. Then, for the imaging performance, the tolerance to the fabrication and assembly of the optical components in conventional confocal microscopes is also investigated by simulations. The simulation results indicate the importance of eliminating the effects of stray light in the microscopic systems.