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The PB smear is a microscopic examination of the blood in which cell components are identified, quantified, and evaluated for abnormalities. The white blood cells are classified as nucleated or nonnucleated cells. The nucleated cells are further subdivided into bands, basophilic stippling, nucleated red cells, and nucleated red cells. A differential count is obtained by examining at least 100 cells, counting all small lymphocytes, granulocytes, and erythrocytes, and then counting large mononuclear cells and platelets (Figures 3.1 and 3.2). This gives the white blood cell count with differential. A white blood cell count in excess of 100,000/mm³ is considered abnormal. Figure 3.3 presents a typical hemogram with leukocytosis, and Figure 3.4 presents a normal smear. The most common abnormalities are:
Leukopenia: Decreased number of leukocytes. Common causes of leukopenia include chemotherapy, radiation, and infection. In some cases, an underlying malignant disease is associated with a decrease in the total number of leukocytes. Leukopenia is indicated by hemoglobin concentration, packed cell volume, and white blood cell count.
Leukocytosis: Increased number of leukocytes.
Lymphocytosis: Increased number of lymphocytes.
Thrombocytopenia: Decreased number of platelets.
Hypogranulocytosis: Decreased number of neutrophils or granulocytes. Causes include drug therapy, malignancy, infection, or nutritional deficiency.
The preparation of blood films is critical for proper microscopic examination. In general, a drop of fresh, properly anticoagulated blood is smeared onto a clean, glass microscope slide. A cover slip or rubber band is used to maintain the shape of the blood smear. Smear preparation is operator dependent and thus needs to be performed carefully and skillfully. The blood film must be wet and should not touch the edges of the slide because this area must be available for scanning. The smear should be examined at 20°-40° and the smear edge should extend beyond the area to be scanned. It is best to stain the smear with Romanowsky-type stains such as May-Grnwald-Giemsa and Wright stains.7 The smear should be dried between 30 and 60 minutes before scanning to ensure proper fixation and air drying. The longer it takes to dry the smear, the more difficult it is to fix and stain the smear. The edges of the smear should be properly dried, because this area should be available for scanning. The edges of the smear should be clean and free of precipitate.7
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