Wright Stain, Modified

Newcomer Supply Beaker

(use: Stronger for auto stainers.)

Product Options:

Part # 1421A 500 ml $28.40 H
Part # 1421B 1 L $46.50 H
Part # 1421C 1 gal $90.50 H

SOLUTION:

500 ml 1 Liter 1 Gallon
Wright Stain, Modified Part 1421A Part 1421B Part 1421C

 

Additionally Needed:

Alcohol, Methanol Anhydrous, ACS Part 12236
Wright Stain Buffer, pH 6.8 Part 1430

 

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Wright Stain, Modified for Smears, provides a concentrated Wright’s formula for differential staining of cell types in peripheral blood smears and bone marrow smears/films. This procedure is applicable for either hand or automated staining processes.

 

METHOD:

Technique: Flat staining rack method
Solutions: All solutions are manufactured by Newcomer Supply, Inc.

 

PRESTAINING PREPARATION:

    1. Prepare within an accepted time frame, a well-made blood smear or bone marrow smear/film per your laboratories protocol, with a focus on uniform cell distribution.
    2. Allow slides to thoroughly air-dry prior to staining.
    3. Filter Wright Stain, Modified prior to use with quality filter paper.
        1. Filter sufficient stain to allow 1 ml of stain per slide.

 

STAINING PROCEDURE:

    1. Place slides on flat staining rack suspended over sink.
    2. Fix by flooding slides with Methanol (Part 12236); 10-30 seconds.
    3. Drain off
    4. Flood each slide with 1 ml of filtered Wright Stain, Modified for 3-5 minutes.
        1. See Procedure Notes #1 and #2.
    5. Retain Wright Stain, Modified on slides.
    6. Directly add 1 ml of Wright Stain Buffer, pH 6.8 (Part 1430) to each slide; agitate gently to mix with retained Wright Stain.
    7. Stain for an additional 6-10 minutes.
    8. Wash well in distilled water; rinse thoroughly in running tap water.
    9. Air-dry slides in a vertical position; examine microscopically.
    10. If coverslip is preferred, allow slides to air-dry and coverslip with compatible mounting medium.

 

RESULTS:

Erythrocytes Pink
Neutrophils Granules – Purple
Eosinophils Granules – Pink
White blood cells Chromatin – Purple
Lymphocytes Cytoplasm – Blue
Monocytes Cytoplasm- Blue
Bacteria Deep Blue

 

PROCEDURE NOTES:

    1. Timings provided are suggested ranges. Optimal times will depend upon staining intensity preference.
    2. Smears containing primarily normal cell populations require minimum staining time; immature cells and bone marrow smears/films may require longer staining times.
    3. The color range of stained cells may vary depending on buffer pH and pH of rinse water.
        1. Alkalinity is indicated by red blood cells being blue-grey and white blood cells only blue.
        2. Acidity is indicated by red blood cells being bright red or pink and lack of proper staining in white blood cells.
        3. If necessary, adjust buffer pH accordingly to 6.8 +/ – 0.2.

 

REFERENCES:

    1. Lillie, R. D., and Harold Fullmer. Histopathologic Technic and Practical Histochemistry. 4th ed. New York: McGraw-Hill, 1976. 747-748.
    2. McPherson, Richard and Matthew Pincus. Henry’s Clinical Diagnosis and Management by Laboratory Methods. 22nd ed. Philadelphia: Elsevier Saunders, 2011. 522-532.
    3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 154-155.
    4. Modifications developed by Newcomer Supply Laboratory.