Histotechnology Technical methods

Enzyme Histochemical Stain

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ACETYL CHOLINESTERASE

SECTIONS

Unfixed cryostat or cold Formol calcium/gum sucrose fixed sections.


SOLUTIONS

a. Incubating solution.

--acetyl thiocholine iodide 5mg

--0.1M acetate buffer (pH.6.0) 6.5ml

--0.1m sodium citrate (2.94g/100ml) 0.5ml

--30mM copper sulphate (0.58g/100ml) 1.0ml

--Distilled water 1.0ml

--5mM potassium ferricyanide (0.165g/100ml) 1.0ml

--Add in order, mixing well at each stage


METHOD

1. Incubate in solution a. at 37ÉC for 15-120 minutes.

2. Rinse in distilled water.

3. Counterstain in Haematoxylin.

4. Dehydrate, clear . Mount sections in DPX


RESULTS

Cholinesterase activity Red/brown.


NOTES

i) Care should be taken when using cholinesterase inhibitors as they are extremely toxic.

ii) Unfixed cryostat sections of rectal biopsy take one hour.

iii) Unfixed cryostat sections of muscle biopsy take one and half to two hours.

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