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A method for measuring the buoyant density of platelets which function normally after the procedure, has been developed and critically compared to published methods.  Following velocity sedimentation into a gradient of polyvinyl pyrrol 1idone-coated colloidal silica particles (Percoll) at physiological pH and osmolarity, 90% (n=10, SD=10) of the whole blood platelet population was recovered with leukocyte contamination of .002% (n=10, SD=.001); erythrocyte contamination was less.  After centrifugation to equilibrium through a second continuous linear Percoll gradient, platelet density showed a Gaussian distribution about a mode of 1.0645 g/ml. (n=13, SD=.0015). Leakage of β thromboglobulin (βTG), lactic dehydrogenase (LDH) and serotonin into the gradients was negligible. Intracellular LDH, serotonin and total protein correlated closely with platelet count, but βTG distribution was skewed towards the denser fractions  Platelets recovered from the second gradient showed normal aggregation patterns and produced thromboxane B2 and secreted βTG when stimulated by thrombin or arachidonic acid. Addition of plasma was necessary to produce ristocetin-and ADP-induced aggregation, and significant radioactivity was not associated with platelets isolated from blood to which 125I had been added  Under physiological conditions, arabinogalactan II (Stractan) caused more spontaneous leakage of α-and dense- granule markers than did Percoll. Platelets taken from different interfaces of a discontinuous Stractan gradient were seen to contain platelets of the full density range when they underwent equilibrium centrifugation in linear continuous Stractan or Percoll gradients.  True buoyant density of minimally altered platelets has been measured by this new technique.

The Density Distribution Of Intact Platelets Following Their Isolation From All Other Blood Constituents