Neutrophils are typically the first immune cells to be deployed to a site of damage or infection and are critical to human health. The multi-step adhesion cascade is a paradigm that describes a series of increasingly adhesive steps that circulating leukocytes make with the underlying endothelium, culminating in their complete arrest from blood flow and subsequent migration towards the affected area. For over a century, microscopes have enabled researchers to visualise leukocyte behaviours such as tethering, rolling, firm adhesion and transendothelial migration (TEM). In only the last few decades, researchers have begun to unravel the molecular mechanisms that control these cellular behaviours. In their article, Aleksandar Ivetic and colleagues (Rahman et al., 2021) show that L-selectin, a cell adhesion molecule that hitherto was only known to regulate neutrophil tethering and rolling, is actively involved in regulating neutrophil TEM. Using FRET, they show that L-selectin co-clusters in cis with PECAM-1 specifically during TEM – serving to expedite movement across TNF-, but not IL-1β-activated endothelial monolayers. This cytokine-specific co-clustering between L-selectin and PECAM-1 is explored in more detail and highlights the diverse involvement of L-selectin in the neutrophil multi-step adhesion cascade.