Two-dimensional (2D) materials with van der Waals stacking have been reported to have extraordinary mechanical and electromechanical properties, which give them revolutionary potential in various fields. However, due to the atomic-scale thickness of these 2D materials, their fascinating properties cannot be effectively characterized in many cases using conventional measurement techniques. Based on typical microscopy techniques such as scanning electron microscopy (SEM), transmission electron microscopy (TEM), and atomic force microscopy (AFM), a range of in situ microscopy techniques have been developed to systematically quantify the mechanical and electromechanical properties of 2D materials. This review highlights the advancements of in situ microscopy techniques for studying elasticity and fracture, adhesion and separation, structural superlubricity, as well as c-axis piezoresistivity and rotation angle-related transport of 2D materials. The methods and results of various microscopy experiments, including nanoindentation using AFM, pressurized bubble tests, self-retraction experiments, pull-to-peel methods and so on, are compared, and their respective advantages and limitations are discussed. Finally, we summarize the current challenges in these microscopy techniques and outline development opportunities.