Platelet labelling Protocol

1. Dilute citrated whole blood 1:10 in PBS.
2. Diluted whole blood (50 ml) was incubated with 5 ml of FITC conjugated monoclonal antibody (CD42a, or CD41 or CD61) and 5 ml PE conjugated Glycophorin-A at RT for 15 mins. This enables separation of platelets from noise and red blood cells.
3. This preparation was then diluted with PBS (300 ml) and analysed by flow cytometry.
4. Instrument settings were FSC and SSC parameters adjusted to LOG settings to enable detection and separation of platelets from electronic noise. An event stop gate was placed around the platelet cloud and a minimum of 10,000 events collected.