A Computer-Assisted Image-Analysis System for Analyzing Polymorphonuclear Leukocyte Chemotaxis in Patients with Diabetes Mellitus

A computer-assisted image-analysis system that precisely tracks the cell movements of up to 50 polymorphonuclear leukocytes (PMNLs) was developed and used to quantitatively measure cellular chemokinesis and chemotaxis in normal individuals and in diabetic patients with hyperglycemia. The PMNLs were tested in Zigmond chambers with or without a gradient of 10-7 M n-formylmethionylleucylphenylalanine (f-Met-Leu-Phe), Cellular movement was recorded on videotape by using a videocamera mounted on the microscope. The videotapes were analyzed by computer programs to calculate the speed and direction of each PMNL at to-sec intervals. Average rates of chemokinesis were 19.6 μm/min without and 25.3 μm/min with f-Met-Leu-Phe. McCutcheon indices, which measure chemotaxis, were 0.01 without and 0.48 with f-Met-Leu-Phe, Similar values were observed in diabetic patients after fasting (average glucose, 217 mg/1OO ml) and 2 hr after glucose challenge (average glucose, 309 mg/1OO ml), These values demonstrate that PMNLs from diabetic patients with hyperglycemia move at normal rates and respond appropriately to f-Met-Leu-Phe.