Intraluminal occlusion of the middle cerebral artery (MCA) is used extensively in cerebral ischemia research. We tested a modified nylon suture in a rat model of middle cerebral artery occlusion (MCAO) under two anesthesia regimens. Sprague-Dawley rats were divided into six groups (Group 1, Poly-L-lysine-coated suture under ketamine/xylazine anesthesia; Group 2, modified suture under ketamine/xylazine anesthesia; Group 3, Poly-L-lysine-coated suture under ketamine/xylazine anesthesia with mechanical ventilation; Group 4, modified suture under ketamine/xylazine anesthesia with mechanical ventilation; Group 5, Poly-L-lysine-coated suture under isoflurane anesthesia; Group 6, modified suture under isoflurane anesthesia) and subjected to 2 hours MCAO. Regional cerebral blood flow (rCBF) was monitored by Laser-Doppler flowmetry. Neurological evaluation and ischemic lesion (TTC stain) were assessed at 24 hr of reperfusion. The total ischemic lesion (sum of areas with lacking and intermediate TTC staining) was similar among all six groups. Compared with a Poly-L-lysine-coated suture technique, the modified suture technique produced a lower rCBF, larger infarct size, smaller variance of infarct size, and greater neurological deficit. In addition, isoflurane significantly reduced infarct size. We conclude that use of this modified suture technique with ketamine/xylazine anesthesia and mechanical ventilation produces a more consistent change in cerebral ischemic damage following MCAO in rats.