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Electron Microscopy Sciences

Technical Data Sheets

Papanicolaou Stain For Membrane Filters

EMS Catalog #26801

Fixation

95% Ethyl Alcohol

Staining

  1. Fixation in 95% ethyl alcohol for 15 minutes.
  2. Two changes of distilled water, 20 dips each.
  3. Stain in Gills Hematoxylin 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

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.

Online Ordering

Gill’s Hematoxylin No 1, Hydrochloric Acid 0.05%, Scott’s Water Solution, Modified Orange G Staining Sol. and Modified EA Staining Solution for Papanicolaou Stain for Membrane Filters are available online from the EMS Catalog. For ordering or product information, click here.