Unfixed Cryostat or cold Formal Calcium Fixed/Gum Sucrose treated sections on Jejunal Biopsy (5-7m)
A. Phosphate - Citrate Buffer
--Stock A: 0.2M Disodium hydrogen orthophosphate 2.83g/100ml
--Stock B: 0.1M Citric acid 2.1g/100ml
Composition of Buffer pH 7.0
--Stock A 82.3ml
--Stock B 17.7ml
B. Incubating Solution
--Sodium Nitrite 0.4ml
--(Add the pararosaniline dropwise to)
--(the Sodium Nitrite until the solution)
--(is corn coloured.)
--pH7 0.1M Citric Acid/Phosphate Buffer 9.2ml
--(Adjust pH to 6.5 after mixing well)
--N:N-Dimethyl formamide 0.5ml
1. Incubate for 1 hour at room temperature, by hot plate.
2. Rinse in distilled water.
3. Immerse in 4% formaldehyde for half an hour.
4. Wash, counterstain in haematoxylin for 30 seconds to 1 minute.
5. Wash, dehydrate, clear and mount in D.P.X.
Sucrase activity - Orange/Red.
NB: Incubating time may be longer depending on the potency of the solution.
NOTE: No responsibility is assumed by The University of Nottingham or the Queens Medical Centre NHS Trust for any injury and/or damage to persons or property as a matter of products liability, negligence or
otherwise, or from any use or operation of any methods, products, instructions or ideas contained in the material herein. It is the users responsibilty to ensure that all procedures are carried out according to appropriate Health and Safety requirements.
© Copyright 1997 University of Nottingham Medical School Division of Histopathology. This page was last built on Mon, May 5, 1997
with Frontier. Thanks for looking in. Comments to James.Lowe@nottingham.ac.uk