May Grunwald/Giemsa for Stain Bone Marrow
EMS Catalog #: 26250-Series
- 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.
- Sections: Zenkers’s or other well-fixed tissue. Start with step #1
Paraffin, 6 microns
- Deparaffinize and hydrate to distilled water.
- 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
- Wash in tap water, 10 minutes and rinse in distilled water for two changes
- Place promptly in Methyl Alcohol, two changes, three minutes each.
- 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. Drai, no rinsing. Alternatively, stain in Working Jenner’s (dilute 1:1 with distilled water) 6 minutes.
- Sections: Stain in Working Giemsa (freshly dilute 2.5ml Giemsa Stock Solution (#26250-02) in 50ml distilled water, do not reuse), 45 minutes
- Smears: Cover for 12 minutes with Working Giemsa (15 drops of Giemsa Stock Solution (#26250-02) to 10 drops of distilled water)
- 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.
- Smears: Differentiate in distilled water, agitating for approximately 5 seconds and checking under the microscope.
- Section only: dehydrate quickly in 95% alcohol and then absolute alcohol, and clear in two changes of xylene.
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.