Platelet counting-development of a reference method and a reference preparation

A reference method has been defined for platelet counting by counting chamber haemocytometry, and a procedure has been developed, using platelet rich plasma (PRP) and a Coulter ZBI counter which provides a reliable, rapid and relatively simple method for platelet counting which is closely comparable to the reference method and can thus be used both as a secondary reference method and a routine method. The optimal method for obtaining PRP from EDTA blood was by augmented sedimentation with Boyum's methyl cellulose-metrizoate mixture at sp. gr. 1.08. This was shown to yield a platelet suspension which reflects closely the platelet content of the original blood. Calibration of electronic counters for platelet counting requires material which is stable and which parallels natural human platelets in size, distribution and other physical characteristics. A suspension of glutaraldehyde-fixed human platelets in glycerol appears to be suitable as a reference preparation. Its method of production is described.