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)



Prof.Dr.D. Tschöpe, Diabetesklini, Herz- und Diabeteszentrum Nordrhein-Westfalen, Universitätsklinik der Ruhr-Universität Bochum, Georgstr.11, D-32545 Bad Oeynhausen, Germany, Tel: +49/5731/972292, Fax: +49/5731/971967
Last Update: Jan 02, 2012