Purpose: To evaluate quantitatively leukocyte dynamics in vivo in the rat retinal microcirculation during ischemia reperfusion injury with the use of acridine orange digital fluorography.
Methods: Retinal ischemia was induced in anesthetized pigmented rats by a temporary ligation of the optic nerve. After 60 minutes of ischemia, leukocyte behavior in the retinal microcirculation was evaluated, with acridine orange digital fluorography--consisting of a scanning laser ophthalmoscope and the fluorescent nuclear dye, acridine orange--during reperfusion at 1, 2, 4, 6, 12, 24, 48, 96, and 168 hours. The obtained images were recorded on videotape and analyzed with a computer-assisted image analysis system.
Results: Rolling leukocytes along the major retinal veins were observed in treated rats during the reperfusion period; no rolling leukocytes were observed in the control rats. The number of rolling leukocytes gradually increased and peaked at 102 +/- 40 cells/minute 12 hours after reperfusion; few rolling leukocytes were observed at 96 hours. The velocity of rolling leukocytes at 12 hours (19.1 +/- 3.5 microns/second; P < 0.05) was significantly lower than that at the other three times. No rolling leukocytes were observed along the arterial walls throughout the experiments. The number of accumulated leukocytes increased as time elapsed, peaked at 931 +/- 187 cells/mm2 24 hours after reperfusion, and decreased thereafter.
Conclusions: Leukocyte dynamics in the retinal microcirculation can be quantitatively evaluated during ischemia reperfusion injury.