Platelet analysis using flowcytometric procedures

'Ex vivo' testing of functional platelet properties using conventional techniques reflects the overall behaviour of the whole platelet population in the sample under investigation. Since many functional aspects depend on ultrastructural constituents which may vary from one cell to another, multiparameter single cell analysis of platelets may be advantageous in providing direct insight into deviations at the cellular level relevant to the pathophysiology of disease states such as bleeding disorders or thrombophilia. Immunolabelling with monoclonal antibodies against membrane antigens has been combined with flowcytometry to provide a standardized, highly specific and sensitive analytical tool. The assay has been optimized for simultaneous two colour fluorescence staining, and this allows the testing of whole blood to provide a quick monitoring method for the differential diagnosis of thrombasthenic diseases like Bernard Soulier's syndrome or Glanzmann's thrombasthenia in which typical staining patterns lack the specific fluorescence for glycoproteins Ib and IIb/IIIa respectively. Also changes in the antigenicity of the outer membrane of activated platelets are detectable with monoclonal antibodies against specific antigenic epitopes such as thrombospondin (a secretion marker) or α-granule and lysosomal proteins (extrusion markers). However, for detection of activated platelets in diseases associated with a prethrombotic state, the procedures for immunolabelling platelets with monoclonal antibodies and instrumental detection sensitivity remain to be optimized. After further development, flowcytometric assays of the functional status of individual platelets may be superior to the measurement of the indirect plasma markers such as platelet factor 4 or β-thromboglobulin for routine diagnosis of the prethrombotic state.