Histotechnology Technical methods

Enzyme Histochemical Stain

Methods Index

Pathology Homepage

PHOSPHORYLASE

SECTIONS

Unfixed cryostat


SOLUTIONS

Add the reagents in the following order. Use unfiltered.

Incubating solution

--0.1M Acetate buffer pH5.9 100ml

--0.1M Magnesium chloride 10ml

--Glucose-1-phosphate 1g

--Glycogen (Oyster/rabbit) 20mg

--ATP salt 50mg

--Sodium fluoride 1.8g

--Ethanol 20ml

--Polyvinyl pyrolidine MW, 44,000 9g

--STORE AT - 20ÉC in 30ml aliquots


METHOD

1. Incubate in solution at 37ÉC for 90 minutes.

2 Fix in ethanol for 3 minutes. AIR DRY.

3 Wash in 1:30 lugols iodine for 5 minutes

4 Mount in 9:1 glycerine jelly/lugols iodine.

RESULTS

Phosphorylase activity - blue/black.


NOTES

The Solution is kept in a closed columbia jar and is frozen after each incubation

Replace when its potency is diminished.

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