Giemsa, Stock

Reagents for this procedure are sold as individual stain solutions and are available for purchase under separate part numbers with storage requirements and expiration date designated per bottle.

Product Options:

Part # Non-Kit

                                                                                     


                               

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

Additionally Needed:

Alcohol, Methanol Anhydrous, ACS Part 12236

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 staining procedure provided below.

 

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 smear to thoroughly air-dry prior to staining.
  3. Fix smear in Methanol for 3-5 minutes.
  4. Air-dry slides in a vertical position.
  5. Prepare Working Giemsa Stain; combine, mix well and filter.
  1. Giemsa Stock Stain Solution            6 ml
  2. Distilled water                                        30 ml
  3. See Procedure Note #1.
  1. Stain in filtered Working Giemsa Stain for 30-45 minutes.
  1. See Procedure Notes #2 and #3.
  1. Wash in distilled water.
  2. Air-dry slides in a vertical position, then examine microscopically.
  3. If coverslip is preferred, allow slides to air-dry and 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. For more intense staining substitute a 1% sodium carbonate solution for distilled water in preparing the Working Giemsa Stain.
  2. The timings provided in this procedure are suggested ranges.  Optimal staining times will depend upon staining intensity preference.
  3. 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.

 

REFERENCES:

  1. Bailey, W. Robert, and Elvyn Scott. Diagnostic Microbiology. 4th ed. St Louis: C. V. Mosby Company, 1974. 394.
  2. Luna, Lee G. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. 3rd ed. New York: Blakiston Division, McGraw-Hill, 1968. 127-128.
  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.