Hi April, Try this Control tissues: liver, kidney and prostate Fixation and Sectioning: Air dried unfixed 8µm frozen sections
Solutions: 1. Substrate solution Naphtol AS-B1 phosphate 0.005g (Fluka 70494 1g) Dimethylfomamide 0.5ml Warning: Suspected Carcinogen – see MSDS 2. Acetate buffer pH 4.8 Sodium acetate 4.8g Distilled water 100ml 0.6% Acetic Acid 14.75ml pH to 4.8 Make up to 1000 ml with distilled water 3. Sodium nitrite: Warning: Toxic – see MSDS Sodium nitrite 1.0g Distilled water 25.0ml 4. Pararosaniline-HCl stock solution. Warning: Suspected Carcinogen – see MSDS Pararosaniline hydrochloride (Basic Fuchsin CI 42510) 0.8g Distilled water 16ml Concentrated Hydrochloric acid 4ml Heat gently, cool to room temperature and filter (store 4C) 5. Incubation solution: Take 0.4ml solution 3 + 0.4ml solution 4 and mix. Stand for 2 minutes and then add to: Substrate Solution (Solution 1) 0.5ml Acetate buffer pH 4.8 25ml Adjust with 0.1M NaOH to pH 4.5-5.0. Procedure: 1. Take air-dried sections and incubate in the incubation solution at 37oC for 1 hour 2. Wash with distilled water and counterstain with Harris's Haematoxylin for 1 minute. 3. Wash in running tap water 4. Dehydrate rapidly through fresh alcohols to xylene and mount. Results: Acid phosphatase activity red Nuclei blue 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 ________________________________________ From: Oler, April via Histonet [histonet@lists.utsouthwestern.edu] Sent: Friday, January 26, 2018 6:26 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Acid Phosphatase Protocol Good afternoon, I am writing to you all because I am in search of Acid Phosphatase muscle protocols that do not utilize sodium barbital. I'm hoping to eliminating sodium barbital from my lab in the near future. If anyone is willing to share their protocol, or advice, I would very much appreciate any assistance. Thank you! April Oler, HT(ASCP) Senior Clinical Technologist IPOX/DIF/Muscle Laboratories Michigan Medicine Ann Arbor, MI ********************************************************** Electronic Mail is not secure, may not be read every day, and should not be used for urgent or sensitive issues _______________________________________________ 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