EMS26801

Components:

  • Gill’S Haematoxylin No 1

  • Hydrochloric Acid 0.05% Solution

  • Scott’s Water Solution

  • Modified Orange G Solution

  • Modified EA Staining Solution

Fixation:

95% Ethyl Alcohol

Staining Procedure:

  1. Fixation in 95% ethyl alcohol for 15 minutes.

  2. Two changes of distilled water, 20 dips each.

  3. Stain in Gills haematoxylin No. 1 for 2-3 minutes.

  4. Wash in two changes of distilled water until the excess stain is removed from the filter.

  5. Place in Hydrochloric Acid 0.05% for 45 seconds or until the filter turns yellow.

  6. Wash in two changes of distilled water, 20 dips each.

  7. Scott’s Tap Water Substitute for 1 minute.

  8. Wash in two changes of distilled water, 20 dips each.

  9. Two changes of 95% ethyl alcohol, 10 dips each.

  10.  Counterstain in Modified OG-6 Staining Solution for 2 to 3 minutes.

  11.  Two changes 95% ethyl alcohol, 1 minute each

  12.  Counterstain further in Modified EA Staining Solution for 2 to 3 minutes.

  13.  Dehydrate in 2 changes each of 95% ethyl alcohol and absolute ethyl alcohol. Clear in 2 changes of Xylene.

  14.  Soak filter in Permount for 15 minutes and mount in Permount.

Stain Results:

Nuclei

Deep Blue to Purple

References:

Gill. G.W., Acta Cytol., 18:300-311, 1974

Diagnostic Cytology by Membrane Filter, Application Bulletin 100, Gelman Instrument Co., Ann Arbor, MI, 1978.