The Düsseldorf- IIIx- Protocol
- Blood Vessel Puncture (arterial or undwelled venous; 1mm - caliber needle;
no stasis)
- Immediate Stabilization (10 Vol.-% Special Medium)
- Second Step Fixation (0.5% Paraformaldehyde)
- Differential Centrifugation (10 Min., 250g; 5 Min., 700g)
Platelet Rich Plasma (PRP)
- Washing
- Blocking Fc- Receptors with 10 Vol.-% Rabbit Serum
- Adjustment of Platelet Count (50000/ul)
- Staining with Monoclonal Antibody Panel (titrated saturating conditions)
- Indirect Fluorescence Labeling with F(ab)2- FITC fragments (titrated saturating
conditions)
- Washing
- Flow Cytometric Analysis (FacScan, approved instrument)
- List Mode Data Processing
Test Evaluation:
1.) Percentage Marker Positive Platelets (qualitative)
2.) Mean Fluorescence Intensity (quantitative)
3.) Activation Index (= 1 x 2)