I have found that that this gives a pleasing tinctorial result: Haematoxylin and Eosin
Principle Traditional HE procedures generally worked poorly on GM sections, with background staining of the plastic embedment and the loosening of sections from glass slides when alcoholic eosin is used. Aqueous solutions tend to stain poorly. Sections are firstly stained with Celestine blue, followed by a regressive Harris's haematoxylin (excess stain is removed from the plastic with acid alcohol). The sections are then counterstained with buffered eosin Y/Phloxine B. Solutions 1. Celestine Blue - Iron Alum 2. Harris's Haematoxylin 3. Buffer for Eosin Y/Phloxine B. Solution A 2.875ml Glacial Acetic acid in 500ml distilled water. Solution B 4.1g Sodium Acetate in 500ml distilled water. 150 ml Solution A + 350 ml Solution B. pH to 4.85. 4. Eosin Y/Phloxine B. Eosin Y (CI 45380) 5g. Phloxine B (CI 45410) 0.5g. Buffer 500ml. Add 2 crystals Thymol. Technique 1. Hydrate sections. 2. Stain in Celestine Blue 15 minutes. 3. Wash in running water. 4. Stain in Harris's Haematoxylin 15 minutes. 5. Wash, differentiate and blue till surrounding plastic is clear. 6. Stain in buffered Eosin for 6 minutes. 7. Wash lightly in running water. 8. Differentiate in alcohol till surrounding plastic is clear. 9. Clear in Xylene and mount. REFERENCES: Castro,M.D., (1985) "A Haematoxylin - Eosin Phloxine Stain for tissues embedded in Glycol Methacrylate" J. Histotechn 8(1); 23-24. Green,G.H.,Kurnein,F., (1981) "Glycol Methacrylate embedding in General Histopathology " ACP Broadsheet 97. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: Keena Leon Guerrero via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Friday, 25 September 2020 8:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] JB4 embedded specimens Hello All, I was wondering if anyone has a good Hematoxylin and Eosin (H&E) staining protocol for JB4 embedded specimens? Thanks! Keena Leon Guerrero _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of NSW Health or any of its entities. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet