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Wright Stain

Wright’s stain is a histologic stain that facilitates the differentiation of blood cell types. It is classically a mixture of eosin (red) and methylene blue dyes. It is used primarily to stain peripheral blood smears, urine samples, and bone marrow aspirates which are examined under a light microscope.

Wright's Stain
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Reagents

The dye may be purchased as a powder which is then mixed to methanol or a ready-made solution may be obtained.

  1. Staining Solution
    Wright’s stain powder = 1.0 gm
    Water free methanol = 400 ml
  2. Phosphate buffer (0.15M, ph 6.5/6.8)
    Potassium dihydrogen phosphate, anhydrous = 0.663 gm
    Disodium hydrogen phosphate, anhydrous = 0.256 gm
    Distilled water = 100 ml

Storage and Shelf life:

Wright’s Stain should be stored at room temperature and protected from light. Under these conditions it has a shelf life of 52 weeks from the date of manufacture.

Procedure:

  • Dip slide for a few seconds in methanol as a fixative step and allow slide to air dry completely.
  • Place slide on a level staining rack and place 1.0 ml of the Wright Stain Solution upon the smear 1 – 3 minutes.
  • Add 2.0 ml distilled water or Phosphate buffer pH 6.5 and let stand twice as long as 1-3 minuts.
  • Rinse stained smear with water or the Phosphate buffer pH 6.5 until the edges show faintly pinkish-red.
  • Wipe the underside of the slide and let the slide air dry in a vertical position.
  • Examine the slide under the microscope.

Results:

Erthrocytesyellowish-red
Polymorphonuclears: Nucleus dark purple
Polymorphonuclears: Granulesreddish-lilac
Polymorphonuclears: Cytoplasmpale-pink
Eosinophiles: Nucleiblue
Eosinophiles: Granulesred to orange-red
Eosinophiles: Cytoplasmblue
Basophiles: Nucleuspurple to dark blue
Basophiles: Granulesvery dark purple
Lymphocytes: Nucleidark purple
Lymphocytes: Cytoplasmsky blue
Plateletsviolet to purple granules

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