Flow cytometric evaluation of platelet–leukocyte conjugate stability over time: methodological implications of sample handling and processing
- 1 January 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Journal of Thrombosis and Thrombolysis
- Vol. 51 (1), 120-128
- https://doi.org/10.1007/s11239-020-02186-5
Abstract
Platelet activation and subsequent aggregation is a vital component of atherothrombosis resulting in acute myocardial infarction. Therefore, quantifying platelet aggregation is a valuable measure for elucidating the pathogenesis of acute coronary syndromes (ACS). Circulating platelet-monocyte conjugates (PMC) as determined by flow cytometry (FCM) are an important measure of in vivo platelet aggregation. However, the influence of sample handling on FCM measurement of PMC is not well-studied. The changes in FCM measurement of PMC with variation in sample handling techniques were evaluated. The stability of PMC concentrations over time with changes in fixation and immunolabeling intervals was assessed. The effect of Time-to-Fix and Time-to-Stain on FCM PMC measurements was investigated in five healthy volunteers. Time-to-Fix (i.e., interval between phlebotomy and sample fixation) was performed at 3, 30, and 60 min. Time-to-Stain (i.e., time of fixed sample storage to staining) was performed at 1, 24, and 48 h. Increasing Time-to-Stain from 1 to 24 or 48 h resulted in lower PMC measures (p < 0.0001). A statistically significant difference in PMC measurement with increasing Time-to-Fix was not observed (p < 0.41). Postponement of sample staining has deleterious effects on the measurement of PMC via FCM. Delays in immunolabeling of fixed samples compromised measurement of PMC by 30% over the first 24 h. Staining/FCM should be completed within an hour of collection.Keywords
Funding Information
- American Heart Association (11CRP7300003)
- Foundation for the National Institutes of Health (1P20 GM103492)
This publication has 33 references indexed in Scilit:
- Association of platelet‐monocyte aggregates with platelet activation, systemic inflammation, and myocardial injury in patients with non‐st elevation acute coronary syndromesClinical Cardiology, 2007
- Flow cytometric analysis of circulating platelet-monocyte aggregates in whole blood: Methodological considerationsThrombosis and Haemostasis, 2007
- Evaluation Of Platelet Function By Flow CytometryPathophysiology of Haemostasis and Thrombosis, 2006
- Platelets in inflammation and atherogenesisJCI Insight, 2005
- Application of Flow Cytometry to Platelet DisordersSeminars in Thrombosis and Hemostasis, 2004
- The clinical utility of flow cytometry in the study of plateletsSeminars in Hematology, 2001
- The clinical utility of flow cytometry in the study of plateletsSeminars in Hematology, 2001
- The Pathogenesis of Coronary Artery Disease and the Acute Coronary SyndromesThe New England Journal of Medicine, 1992
- The Pathogenesis of Coronary Artery Disease and the Acute Coronary SyndromesThe New England Journal of Medicine, 1992
- Towards a better definition of human leucocyte surface moleculesImmunology Today, 1989