Whole blood flow cytometry protocol for the assessment of platelet phenotype, function, and cellular interactions

Abstract

Platelets are a key component of the hemostatic system and their roles in inflammation via interactions with leukocytes have also gained attention in recent years. Changes in platelet phenotype and function can cause bleeding and/or thrombosis and, as such, monitoring platelet-specific changes is crucial to assessing hemostasis in the clinical setting. Currently, available platelet function tests such as platelet aggregometry and thromboelastography require a large volume of blood, which is a major limitation for the pediatric population. Whole blood flow cytometric analysis of platelets is increasingly utilized in recent years, primarily due to the sensitivity of this method, but also because it only requires a small amount of blood with minimal sample manipulation. We have developed a whole blood flow cytometry methodological approach that enables the assessment of platelet phenotype, function, and their interactions with monocytes and neutrophils.

Keywords:

• Flow cytometry
• platelet function
• platelet-leukocyte aggregates
• whole blood

Acknowledgements

The authors would like to thank Dr. Matthew Burton and his team from the Murdoch Children’s Research Institute Flow Cytometry and Imaging facility for their assistance in troubleshooting the flow cytometry assays. MDL is an ISAC IMI Scholar. We acknowledge funding from the Australian National Health and Medical Research Council, grant 1129317.

Conflicts of Interest

The authors have no conflicts of interest to disclose.

Supplementary material

Supplemental data for this article can be accessed on the publisher’s website.

Additional information

Funding

This study was funded by National Health and Medical Research Council (NHMRC), [Grant Number 1129317].