Twistronics, an emerging field focused on exploring the unique electrical properties induced by twist interface in graphene multilayers, has garnered significant attention in recent years. The general manipulation of twist angle depends on the assembly of van der Waals (vdW) layered materials, which has led to the discovery of unconventional superconductivity, ferroelectricity, and nonlinear optics, thereby expanding the realm of twistronics. Recently, in situ tuning of interlayer conductivity in vdW layered materials has been achieved based on scanning probe microscope. In this Perspective, the advancements in in situ twistronics are focused on by reviewing the state-of-the-art in situ manipulating technology, discussing the underlying mechanism based on the concept of structural superlubricity, and exploiting the real-time twistronic tests under scanning electron microscope (SEM). It is shown that the real-time manipulation under SEM allows for visualizing and monitoring the interface status during in situ twistronic testing. By harnessing the unique tribological properties of vdW layered materials, this novel platform not only enhances the fabrication of twistronic devices but also facilitates the fundamental understanding of interface phenomena in vdW layered materials. Moreover, this platform holds great promise for the application of twistronic-mechanical systems, providing avenues for the integration of twistronics into various mechanical frameworks.