Wright Stain Method

Created by Kathleen Patrick, Modified on Thu, 31 Oct at 1:59 PM by Kathleen Patrick

EMS26060

Fixation:

Streak thin (approximately one cell thick) smears across a sterile slide by means of a second slide or cover glass. Air dry quickly.

Staining:

Place 1.0ml of the Wright Stain Solution upon the smear 1 – 3 minutes.
Add 2.0ml distilled water or Phosphate buffer pH 6.5 and let stand twice as long as in step 1.
Rinse stained smear with water or the Phosphate buffer pH 6.5 until the edges show faintly pinkish-red.
For a Giemsa appearance stain 10 minutes in one volume Wright Stain and 4 volumes Phosphate buffer, pH 6.5.
Blot dry very carefully. Stain may be adjusted by further dilution or in the timing of either before or after dilution in the above procedure.

Results:

Erthrocytes	yellowish-red
Polymorphonuclears: Nucleus	dark purple
Polymorphonuclears: Granules	reddish-lilac
Polymorphonuclears: Cytoplasm	pale-pink
Eosinophiles: Nuclei	blue
Eosinophiles: Granules	red to orange-red
Eosinophiles: Cytoplasm	blue
Basophiles: Nucleus	purple to dark blue
Basophiles: Granules	very dark purple
Lymphocytes: Nuclei	dark purple
Lymphocytes: Cytoplasm	sky blue
Platelets	violet to purple granules

References:

Conn, J I Biological Stains, 8th Ed.,Williams and Wilkins Co. Baltimore, 1969.

Clark, G (ed.) Staining Procedures, Williams and Wilkens Co., Baltimore, 3rd Ed., c 1972, p. 122