Surface expression of P-selectin is known to be a marker of platelet activation in patients with acute coronary syndromes. However, direct comparisons of flow cytometer data may be obscured by differences in methodology, artifactual platelet activation during washing procedures, choice of antibodies, absence of control measurements, and possible observer bias due to unblinded data collection. We sought to test the hypothesis that the model of flow cytometer represents another variable affecting P-selectin measurements. Platelet P-selectin in whole blood was measured by FACScan (Becton Dickinson, Inc., San Diego, CA, USA) or EPICS XL (Coulter Corporation, Hialeah, FL, USA) flow cytometry in 338 patients presenting with chest pain to the emergency departments of three community hospitals as part of a multicenter diagnostic trial. Platelet expression of P-selectin (% of cell positivity) was consistently higher for each discharge diagnosis when measured with FACScan flow cytometer (13.2+/-4.1 for myocardial infarction, 10.0+/-3.6 for unstable angina, 9.9+/-3.5 for heart failure, 4.7+/-0.1 for gastrointestinal illness, and 6.3+/-0.7 for patients with noncardiac chest pain) when compared with results obtained from the EPICS XL instrument (2.4+/-0.2, 2.5+/-0.2, 2.5+/-0.1, 1.8+/-0.1, and 2.3+/-0.1 respectively, p=0.0001 for all groups). This study reveals marked discrepancies in the level of platelet P-selectin measurement based exclusively on the model of flow cytometer used. If P-selectin is to become a diagnostic tool for differentiating an etiology of chest pain, standardized measurements must be defined for each model of flow cytometer.