Products

Wright Stain, Buffered

 

For staining procedure, click here.

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Part # Non-Kit

                                                                                    

SOLUTION: 500 ml  1 Liter 1 Gallon
Wright Stain Solution, Buffered Part 1422A Part 1422B Part 1422C

Additionally Needed:

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

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

 

APPLICATION:

Newcomer Supply Wright Stain, Buffered for Smears provides a quick staining technique, used for differential staining of cell types in peripheral blood smears as well as bone marrow smears/films.

 

METHOD:

Technique: Coplin jar or flat staining rack method

Solutions: All solutions are manufactured by Newcomer Supply, Inc.

 

STAINING PROCEDURE:

  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 Solution, Buffered prior to use.
  4. Prepare 25% Aqueous Methanol Rinse; combine and mix well.
  1. Distilled water                           30 ml  or  3 ml
  2. Methanol                                     10 ml  or  1 ml
  1. Coplin Jar Method: See Procedure Notes #1 and #2.
  1. Fix smears in methanol for 15 seconds.
  2. Stain in Wright Stain Solution, Buffered for 1-2 minutes.
  3. Place smear directly into Wright Stain Buffer Solution, pH 6.8, 1 to 4 minutes.  Do Not Agitate!
  4. Dip smears in 25% Aqueous Methanol Rinse for 1 second.
  5. Rinse in distilled water.
  6. Air-dry slides in a vertical position, then examine microscopically.
  7. If coverslip is preferred, allow slides to air-dry and coverslip with compatible mounting medium.
  1. Flat Staining Rack Method: See Procedure Notes #1 and #2.
  1. Place slides on a flat staining rack suspended over a sink.
  2. Fix smear by flooding slide with methanol for 15 seconds.
  3. Shake off excess methanol; apply 1 ml Wright Stain Solution, Buffered to each slide for 1 minute.
  4. Retain Wright Stain Solution, Buffered on slides, directly add 2 ml of Wright Stain Buffer Solution, pH 6.8 to each slide; gently agitate to mix. Stain for an additional 3 minutes.
  5. Flood smears with 25% Aqueous Methanol Rinse for 1 second.
  6. Rinse in distilled water.
  7. Air-dry slides in a vertical position, then examine microscopically.
  8. 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. The timings provided in this procedure are suggested ranges.  Optimal staining times will depend upon staining intensity preference.
  2. Smears containing primarily normal cell populations require minimum staining time; immature cells may require a longer staining time. Bone marrow smears/films may also require a longer staining time.
  3. The color range of the stained cells may vary depending upon the pH of the buffer and the pH of the rinse water used.
  1. Alkalinity is indicated by red blood cells being blue-grey and white blood cells only blue.
  2. Acidity is indicated by red blood cell 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.