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)

PD Dr.D.Tschöpe, Tel: +49/211/3382-665, Fax: +49/211/3382-665, E-mail: tschoepe@dfi.uni-duesseldorf.de, Arb.Gruppe Zelluläre Hämostase und Klinische Angiologie, Diabetes Forschungsinstitut an der Heinrich Heine Universität Düsseldorf, Auf'm Hennekamp 65, D-40225 Düsseldorf, Germany
Last Update: May 9, 1996

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CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories and distributed free of charge as an educational service to the cytometry community. If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL, Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu EMAIL robinson@flowcyt.cyto.purdue.edu