Wolbach's Giemsa Method
EMS Catalog #: 26132-Series
Fixation:
Russell’s Zenker Fixative (#64123-05) or other well fixed tissues.
Sections:
Paraffin at 6 µm sections
Staining:
- Deparaffinize and hydrate to distilled water.
- If Zenker fixed, remove “Zenker’s crystals” by placing in Gram’s Iodine (#26131-04) or Lugol’s Iodine (#26131-4A) for 15 minutes. Rinse in water and clear in Sodium Thiosulfate, 5% (#26131-05) for three minutes. Wash in running water for 15 minutes.
- Rinse in distilled water and stain in Working Giemsa Solution (#26132-1A) overnight.
Working Giemsa Solution may be made from the stock solution (#26132-01) by mixing
- Giemsa Stock Solution - 1.25ml
- Methanol - 1.5ml
- Distilled Water - 50.0ml
NOTE: Giemsa stain colors more effectively in tissue at an acid pH. If this has not occurred in the preparation or decalcification steps, wash in acid alcohol, then begin stain
- Differentiate in Rosin Alcohol Working solution (#26132-2A) until the sections are purplish-pink color. Check under microscope.
- Dehydrate in two changes of absolute alcohol and clear in two changes of xylene.
- Mount with Permount (#17986-01).
Stain Results:
| Nuclei, Bacteria | Blue |
| Rickettsia | Purple |
| Collagen, other tissue elements | Pink to rose |
References:
Wolbach, S.B., Todd, J.L. and Palfrey, F.W., The Etilogy of Pathology Typhus, Harvard University Press, Cambridge, MA., p.13-14, c. 1922
Luna, L.G., (ed) Manual of Histologic Staining Methods of the AFIP, 3rd edition, McGraw-Hill, New York, p. 119, c. 1968.
Online Ordering
Wolbach's Giemsa Method is available online from the EMS Catalog. For ordering or product information, click here.
