Adhesion of leukocytes (L) to microvascular endothelium (E) is a required step in the L-E interaction leading to tissue injury in ischemia-reperfusion. To assess the optimum period for therapy aimed at ameliorating negative effects of this required step, we investigated the time course of L-E adhesion in the hamster cheek pouch using 2 hr of ischemia and 1 hr of reperfusion in our model of I-R injury (Am. J. Physiol-261: 1626, 1991). Leukocytes adhering (stationary for > or = 30 sec) to postcapillary venules (15-30 microns in diameter) were counted after labeling with acridine orange. Prior to the induction of ischemia, there were no significant differences in the number of adherent leukocytes in each area chosen for study (1.9 +/- 0.6 vs 2.0 +/- 0.3; mean number of leukocytes/100-microns vessel length +/- SD). After 10 and 20 min of reperfusion there was no significant difference in leukocyte adhesion in the ischemic area relative to the control (2.7 +/- 0.5 vs 2.8 +/- 0.8, and 5.3 +/- 2.8 vs 2.4 +/- 0.6, respectively). Leukocyte adherence increased significantly after 30 min of reperfusion and remained elevated at 1 hr of reperfusion in the postischemic area relative to the nonischemic control area (7.8 +/- 1.3 vs 3.6 +/- 0.6, and 8.3 +/- 0.8 vs 4.1 +/- 0.6, respectively; P < 0.01). Leukocyte adhesion in the postischemic area after 30 min reperfusion was not significantly different from the adhesion at the end of 1 hr reperfusion. These data suggest that (1) peak leukocyte adhesion occurs after 30 min of normal reperfusion and (2) postischemic therapeutic intervention may be most beneficial when instituted within this early time period.