Background
Flow cytometry is an important technique for understanding multiple aspects of blood platelet biology. Despite the widespread use of the platform for assessing platelet function, the optimisation and careful consideration of pre-analytical conditions, sample processing techniques and data analysis strategies should be regularly assessed. When set up and designed with optimal conditions it can ensure the acquisition of robust and reproducible flow cytometry data. However, these parameters are rarely described despite their importance.
Objectives
We aimed to characterise the effects of several pre-analytical variables on the analysis of blood platelets by multiparameter fluorescent flow cytometry.
Methods
We assessed anticoagulant choice, sample material, sample processing and storage times on four distinct and commonly used markers of platelet activation including fibrinogen binding, expression of CD62P and CD42b, and phosphatidylserine exposure.
Results
The use of sub-optimal conditions led to increases in basal platelet activity and reduced sensitivities to stimulation, however the use of optimal conditions protected the platelets from artefactual stimulation and preserved basal activity and sensitivity to activation.
Summary
The optimal pre-analytical conditions identified here for the measurement of platelet phenotype by flow cytometry suggests a framework for future development of multiparameter platelet assays for high quality datasets and advanced analysis.
ORCID: 
CORE (COnnecting REpositories)
CORE (COnnecting REpositories)