Giemsa Stock Stain

Dilute with water or buffer.

Product Options:

Part # 1120A500 ml $63.50H
Part # 1120B1 L $81.70H

          

SOLUTION: 500 ml 1 Liter
Giemsa Stock Stain Part 1120A Part 1120B

 

Additionally Needed:      

Alcohol, Methanol Anhydrous, ACS Part 12236
Phosphate Buffer, pH 7.0 Part 1331

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

 

APPLICATION:

Newcomer Supply Giemsa Stain is a simple one-step method designed to demonstrate differential staining of cells types in peripheral blood smears and bone marrow smears/films as well as a method for detecting rickettsia, bacteria and parasites.

 

METHOD:

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

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

STAINING PROCEDURE:

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.

  1. Proceed with either the thin or thick smear/film staining method.

 

Thin Smear/Film Staining Method: See Procedure Notes #1 and #2.

  1. Allow smear to thoroughly air-dry prior to staining.
  2. Fix smear in Methanol: 1-2 minutes.
  3. Air-dry slides in a vertical position.
  4. Prepare fresh 1:20 Working Giemsa Stain; combine and mix well.
    1. Giemsa Stock Stain                           2 ml
    2. Phosphate Buffer, pH 7.0 (Part 1331)    40 ml
  5. Stain in Working Giemsa Stain for 20-30 minutes.
  6. Rinse briefly in Phosphate Buffer, pH 7.0 or distilled water.
  7. Air-dry slides in a vertical position.
  8. If coverslip is preferred, allow slides to air-dry; coverslip with compatible mounting medium.

 

Thick Smear/Film Staining Method: See Procedure Notes #1 and #2.

  1. Allow smear to thoroughly air-dry prior to staining; several hours or overnight.
  2. Proceed directly to stain; do not place in fixative.
  3. Prepare fresh 1:50 Working Giemsa Stain; combine and mix well.
    1. Giemsa Stock Stain                           1 ml
    2. Phosphate Buffer, pH 7.0 (Part 1331)    50 ml
  4. Stain in Working Giemsa Stain for 50 minutes.
  5. Rinse briefly in Phosphate Buffer, pH 7.0 or distilled water.
  6. Air-dry slides in a vertical position.
  7. If coverslip is preferred, allow slides to air-dry; coverslip with compatible mounting medium.

 

RESULTS:

Erythrocytes Orange - pink to rose
Platelets Red to purple granules with blue halo

Granulocytes

Neutrophils Nucleus - Dark blue to violet
Cytoplasm - Pink
Granules - Purple to lilac
Eosinophils Nucleus - Blue
Granules - Orange to pink
Basophils Nucleus - Deep blue to violet
Granules - Deep blue to violet

Mononuclear Cells                 

Lymphocytes Nuclei - Deep blue to violet
Cytoplasm - Light blue
Monocytes Nuclei - Light blue/purple
Cytoplasm - Pale gray/blue
Mast cells Nuclei - Deep blue to violet
Granules - Deep blue-violet
Malarial parasites Nucleus - Red chromatin dot
Cytoplasm – Blue
Rickettsia Bluish purple
Bacteria Blue

 

PROCEDURE NOTES:

  1. The timings provided are suggested ranges.  Optimal staining times will depend upon smear/film thickness and preference of stain intensity.
  2. Smears/films containing primarily normal cell populations require minimum staining time.
    1. Immature cells may require a longer staining time.
    2. Bone marrow smears/films may require a longer staining time.

 

REFERENCES:

  1. Bailey, W. Robert, and Elvyn Scott. Diagnostic Microbiology. 4th ed. St Louis: C. V. Mosby Company, 1974. 394.
  2. Garcia, Lynne Shore. Diagnostic Medical Parasitology. 5th ed. Washington DC: ASM Press, 2007. 888-889.
  3. McPherson, Richard and Matthew Pincus. Henry’s Clinical Diagnosis and Management by Laboratory Methods. 22nd ed. Philadelphia:  Elsevier Saunders, 2011. 522-531.
  4. Modifications developed by Newcomer Supply Laboratory.