May Grunwald/Giemsa for Stain Bone Marrow

EMS Catalog #: 26250-Series 

Fixation:

1. Smears: Draw a curetted fragment lightly across a sterile slide in a serpentine path. Don’t allow the streak to dry completely. Start with step #4.
2. Sections: Zenkers’s or other well-fixed tissue. Start with step #1

Sections:

Paraffin, 6 microns

Staining Procedures:

1. Deparaffinize and hydrate to distilled water.
2. If use Zerker’s contains mercury, remove mercuric chloride crystals with Lugol’s or Gram’s Iodine solution, clear with Sodium Thiosulfate, 5% for 3 minutes. If Zenker’s use is contained no mercury, go to step #4
3. Wash in tap water, 10 minutes and rinse in distilled water for two changes
4. Place promptly in Methyl Alcohol, two changes, three minutes each.
5. Stain with May Grunwald Solution (#26250-01) or Jenner Solution (#26250-1A) 3 minutes. Add an equal volume of distilled water and allow to stand one minutes. Drain, no rinsing. Alternatively, stain in Working Jenner’s (dilute 1:1 with distilled water) 6 minutes.
6.  
1. Sections: Stain in Working Giemsa (freshly dilute 2.5 ml Giemsa Stock Solution (#26250-02) in 50 ml distilled water, do not reuse), 45 minutes
2. Smears: Cover for 12 minutes with Working Giemsa (15 drops of Giemsa Stock Solution (#26250-02) to 10 drops of distilled water)
7.  
1. Sections: Differentiate each slide individually in Acetic Acid, 1% with gentle agitation, checking often under the microscope until nuclei are well defined. Rinse quickly in distilled water.
2. Smears: Differentiate in distilled water, agitating for approximately 5 seconds and checking under the microscope.
8. Section only: dehydrate quickly in 95% alcohol and then absolute alcohol, and clear in two changes of xylene.
9. 1. Sections: Mount with Permount (#17986-01) or DPX (#13510)
   2. Smears: Blot with fine grain filter paper and mount.

Stain Results:

References:

Beck, R.C., Laboratory Manual of Hematological Technique. W.B. Saunders & Co., Philadelphia, c. 1938. pp233-236

Luna, L.G., (ed) Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology, 3rd ed. McGraw-Hill, New York, c. 1968, pp212-122.

Clark, G., (ed.). Staining Procedures, 3rd ed., Williams & Wilkins, Baltimore, pp. 131-132, c. 1973.

 Strumia, M.M., J. Lab Clin. Med, 21:930-934, 1935-1936.

Product Information

May Grunwald/Giemsa for Stain Bone Marrow