Previous studies have shown that cryopreservation of normal platelets induces a reduction in the contents of secretion granules, the generation of thromboxane B2, and aggregability. The present study investigates whether these changes in the total population occur to the same extent in four size-dependent subpopulations with mean platelet volumes of 4.2, 5.7, 8.0, and 11.1 μm3, obtained by counterflow centrifugation. Cryopreservation reduced the contents of the alpha granule markers and the generation of thromboxane B2 in the platelets from the four fractions to the same extent as in the platelets from the total suspension. Maximal aggregation of the platelets in response to collagen was measured by optical aggregation. The average decrease in light transmission after freezing was 47 ± 3 percent (SEM) for the platelets in the total population, 40 ± 3 percent for the largest platelets, and 65 ± 5 percent for the smallest platelets, which indicates that aggregability was better preserved in the larger platelets than in the smaller cells. It is possible that, in the smallest platelets, a decrease in thromboxane generation of approximately 70 percent becomes rate-limiting for aggregation. Further improvements in the clinical use of freeze-preserved platelets may be sought in the preparation of concentrates with relatively high counts of large platelets.