Flow cytometric assessment of platelet function in patients with peripheral arterial occlusive disease Spencer W. Galt, MD, Martha D. McDaniel, MD, Kenneth.

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Presentation transcript:

Flow cytometric assessment of platelet function in patients with peripheral arterial occlusive disease Spencer W. Galt, MD, Martha D. McDaniel, MD, Kenneth A. Ault, MD, Jane Mitchell, BS, Jack L. Cronenwett, MD Journal of Vascular Surgery Volume 14, Issue 6, Pages 747-756 (December 1991) DOI: 10.1067/mva.1991.33419 Copyright © 1991 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Sample output from the flow cytometer, examining peripheral venous blood fixed in 2% paraformaldehyde then stained with PLT-1/FITC and biotin S12/streptavidin-phycoerythrin, (a) immediately after phlebotomy or (b) after 5 minutes' stimulation with 20 μmol/L ADP. Previous gating on FITC fluorescence ensures that all particles in both plots are platelets. Log phycoerythrin fluorescence, on the ordinate, is proportional to S12 binding; log forward-angle light scatter, on the abscissa, is proportional to particle size. Quadrants are established by the use of appropriate controls. The line at x = 191 is the size threshold; particles falling below this value are single platelets, whereas those exceeding it are aggregates. The line at y = 5.05 is the log phycoerythrin fluorescence threshold; particles above it exhibit greater phycoerythrin fluorescence than nonspecifically stained platelets. By summing the percent of total events that occur in the upper two quadrants, the total number of platelet-containing particles that are “GMP-140 positive” is determined. Similarly, by summing the percent of total events that occur in the two quadrants to the right, the percent of platelet aggregates is determined. In (a) there are fewer GMP-140 positive platelets and fewer aggregates than in (b), where secretion and aggregation have occurred in response to ADP stimulation. Journal of Vascular Surgery 1991 14, 747-756DOI: (10.1067/mva.1991.33419) Copyright © 1991 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Percent of platelets binding S12 after exposure of whole blood to increasing concentrations of ADP for 5 minutes in vitro. Data are mean ± SEM. Journal of Vascular Surgery 1991 14, 747-756DOI: (10.1067/mva.1991.33419) Copyright © 1991 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Percent platelet aggregates after exposure of whole blood to increasing concentrations of ADP for 5 minutes in vitro. Data are mean ± SEM. *p < 0.05 versus controls; †p = 0.002 versus controls, by ANOVA. Journal of Vascular Surgery 1991 14, 747-756DOI: (10.1067/mva.1991.33419) Copyright © 1991 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions