Intravital imaging of neutrophil recruitment in intestinal ischemia-reperfusion injury

Highlights: • Two-photon laser scanning microscopy is a powerful tool for intravital imaging. • Dynamic neutrophil recruitment can be observed during intestinal I/R. • Imaging reveals the villus height and the diameter of crypt in reperfused ileum. Neutrophils are known to be key players in innate immunity. Activated neutrophils induce local inflammation, which results in pathophysiologic changes during intestinal ischemia-reperfusion injury (IRI). However, most studies have been based on static assessments, and few have examined real-time intravital neutrophil recruitment. We herein report a method for imaging and evaluating dynamic changes in the neutrophil recruitment in intestinal IRI using two-photon laser scanning microscopy (TPLSM).