Papanicolaou Stain for Membrane Filters

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.

Results:

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.