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)
Dimethylfomamide0.5ml
Warning: Suspected Carcinogen – see MSDS
2. Acetate buffer pH 4.8
Sodium acetate 4.8g
Distilled water 100ml
0.6% Acetic Acid14.75ml
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
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