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Platelet Count

Overview :

Blood collection and storage

Platelet counts use a freshly-collected blood specimen to which a chemical called EDTA has been added to prevent clotting before the test begins. About 5 mL of blood are drawn from a vein in the patient's inner elbow region. Blood drawn from a vein helps to produce a more accurate count than blood drawn from a fingertip. Collection of the sample takes only a few minutes.

After collection, the mean platelet volume of EDTA-blood will increase over time. This increase is caused by a change in the shape of the platelets after removal from the body. The changing volume is relatively stable for a period of one to three hours after collection. This period is the best time to count the sample when using electronic instruments, because the platelets will be within a standard size range.

Counting methods

Platelets can be observed in a direct blood smear for approximate quantity and shape. A direct smear is made by placing a drop of blood onto a microscope slide and spreading it into a thin layer. After staining to make the various blood cells easier to see and distinguish, a laboratory technician views the smear through a light microscope. Accurate assessment of the number of platelets requires other methods of counting. There are three methods used to count platelets; hemacytometer, voltage-pulse counting, and electro-optical counting.

HEMACYTOMETER COUNTING. The microscopic method uses a phase contrast microscope to view blood on a hemacytometer slide. A sample of the diluted blood mixture is placed in a hemacytometer, which is an instrument with a grid etched into its surface to guide the counting. For a proper count, the platelets should be evenly distributed in the hemacytometer. Counts made from samples with platelet clumping are considered unreliable. Clumping can be caused by several factors, such as clotting before addition of the anticoagulant and allowing the blood to remain in contact with a capillary blood vessel during collection. Errors in platelet counting are more common when blood is collected from capillaries than from veins.

ELECTRONIC COUNTING. Electronic counting of platelets is the most common method. There are two types of electronic counting, voltage-pulse and electro-optical counting systems. In both systems, the collected blood is diluted and counted by passing the blood through an electronic counter. The instruments are set to count only particles within the proper size range for platelets. The upper and lower levels of the size range are called size exclusion limits. Any cells or material larger or smaller than the size exclusion limits will not be counted. Any object in the proper size range is counted, however, even if it isn't a platelet. For these instruments to work properly, the sample must not contain other material that might mistakenly be counted as platelets. Electronic counting instruments sometimes produce artificially low platelet counts. If a platelet and another blood cell pass through the counter at the same time, the instrument will not count the larger cell because of the size exclusion limits, which will cause the instrument to accidentally miss the platelet. Clumps of platelets will not be counted because clumps exceed the upper size exclusion limit for platelets. In addition, if the patient has a high white blood cell count, electronic counting may yield an unusually low platelet count because white blood cells may filter out some of the platelets before the sample is counted. On the other hand, if the red blood cells in the sample have burst, their fragments will be falsely counted as platelets.

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