The cellular and molecular mechanisms underlying the establishment of vascular connections between primary and lateral roots have recently gained significant attention. Here, I present a protocol to visualize and quantify xylem connection defects during lateral root development. I describe steps for employing stains to infer whether the defects observed in xylem bridges are associated with alterations in the xylem differentiation program, including programmed cell death and secondary cell wall deposition. For complete details on the use and execution of this protocol, please refer to Blanco-Touriñán et al. (2023) and Ursache et al. (2018). ¹ ^, ² .