MXenes, a family of 2D transition metal carbides, nitrides, and carbonitrides, have shown promising performance in various applications such as electromagnetic interference shielding, chemical sensing, and energy storage. To enhance their electronic conductivity, researchers have focused on tailoring the surface chemistry of MXenes. This study investigates the correlation between MXene surface de-functionalization and increased electronic conductivity through in situ vacuum annealing, electrical biasing, and spectroscopic analysis within the transmission electron microscope (TEM). The results show that de-intercalation and surface de-functionalization, particularly the loss of –OH and –F terminations, lead to increased conductivity. Additionally, intercalation can induce transitions between metallic and semiconductor-like transport, with inter-flake effects causing negative temperature-dependence of resistance. These findings provide a foundation for the development of intercalation- and termination-engineered MXenes, which could lead to improved electronic conductivity and the realization of semiconducting, magnetic, and topologically insulating MXenes.MXenes, a family of 2D transition metal carbides, nitrides, and carbonitrides, have shown promising performance in various applications such as electromagnetic interference shielding, chemical sensing, and energy storage. To enhance their electronic conductivity, researchers have focused on tailoring the surface chemistry of MXenes. This study investigates the correlation between MXene surface de-functionalization and increased electronic conductivity through in situ vacuum annealing, electrical biasing, and spectroscopic analysis within the transmission electron microscope (TEM). The results show that de-intercalation and surface de-functionalization, particularly the loss of –OH and –F terminations, lead to increased conductivity. Additionally, intercalation can induce transitions between metallic and semiconductor-like transport, with inter-flake effects causing negative temperature-dependence of resistance. These findings provide a foundation for the development of intercalation- and termination-engineered MXenes, which could lead to improved electronic conductivity and the realization of semiconducting, magnetic, and topologically insulating MXenes.