May-Grunwald/Geimsa Combined Stain For Bone Marrow
- 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: Zenker’s (#64123-05) or other well-fixed tissue. Start with step 1
Paraffin, 6 microns
- Deparraffinize and hydrate to distilled water
- Remove mercuric chloride crystals with Lugol’s or Gram’s Iodine solution with Sodium Thiosulfate, 5% for three minutes.
- Wash in tap water, 10 minutes and rinse in distilled water for two changes.
- Place promptly in Methyl Alcohol, two changes, three times each.
- Stain with May-Grunwald Solution or Jenner Solution 3 minutes. Add an equal volume of distilled water and allow standing one minute. Drain, no rinsing. Alternatively stain in working Jenner’s (dilute 1:1 with distilled water, 6 minutes.
- a. Sections: Stain in working Giemsa (freshly stain in working Giemsa, by diluted 2.5ml Giemsa Stock Solution (#15940)
in 50 ml distilled water, do not re-use), 45 minute
b. Smears: Cover for 12 minutes with working Giemsa (15 drops of Giemsa Stock (#15940) to 10 drops of distilled water)
- a. 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
b. Smears: Differentiate in distilled water, agitating for approximately 5 seconds and checking under the microscope.
- (Section only) Dehydrated quickly in 95% alcohol and absolute alcohol and clear in two changes of xylene.
- a. Sections: Mount with Permount (#17986-01)
or other synthetic mount and
b. Smears: Blot with fine grain filter paper and mount.
|Pink – Rose||Cytoplasm|
Tones are redder than when stained with Giemsa alone.
Beck, R.C., Laborotory Manual of Hematological Technique, W.B. Saunders & Co., Philadelphia, c1938, pp233 - 236
Clark, G., (ed.), Staining Procedures, 3rd Edition, William & Wilkins, Baltimore, pp131 & 132, c 1973.
Strumia, M.M., J. Lab. Clin. Med., 21:930-934, 1935 – 1936.
Luma, L.G., (ed.) Manual of Histologic Staining Method of the AFIP, 3rd edition, McGraw-Hill, NY, pp 121 & 122, c1968.