Histotechnology Technical methods

Enzyme Histochemical Stain

Methods Index

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LACTASE

(Indigogenic procedure according to LOjda et al 1979)

SECTIONS

Unfixed cryostat or cold Formol calcium/gum sucrose fixed, jejunal biopsy.


SOLUTIONS

a. Incubation solution

5-Bromo-4-chloro-3-indoxyl-B D fucoside 5mg 5-Bromo-4-chloro-3-indoxyl-B D fucopyranoside 5mg

Dissolve in

--N N Dimethylformamide 0.5ml

--0.1M citric acid phosphate buffer pH 6.0 10ml --1.65% potassium ferricyanide (50Mmol/L) 0.83ml

--2.11% potassium ferrocyanide (50Mmol/L) 0.83ml


METHOD

1. Incubate sections in the medium at 37ÉC for 1 hour in a closed container to avoid evaporation.

2. Rinse in distilled water.

3. Fix in 4% formaldehyde for 5 minutes at room temperature.

4. Rinse in distilled water.

5. Counterstain in Mayers carmalum for 5 minutes.

6. Was. Dehydrate, clear . Mount sections in DPX


RESULTS

Enzyme activity - Turquoise

NOTES

The medium is expensive but can be used repeatedly. After incubation it can be stored in a closed vial at -20ÉC. The potency of the incubation solution lessens with use and therefore sections may increasing lengths of time in the solution as it ages.

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