Histotechnology Technical methods

Enzyme Histochemical Stain

Methods Index

Pathology Homepage

CYTOCHROME OXIDASE TECHNIQUE

TISSUE

Cryostat sections (Fresh), Muscle Biopsy.

INCUBATING SOLUTION

  • Catalase (20ug/ml) 1ml
    • (4mg in 10ml, remove 2.5ml and make up to 50ml in dist. H20).
  • Cytochrome c (Type 2) 10mg
  • 0.1m phosphate buffer pH 7.4 9ml
  • Add 3,3' diaminobenzidine tetra hydrocholoride (DAB) 5mg
  • Adjust pH to 7.4 before use.

METHOD

  1. Incubate sections at room temperature for 2 hours.
  2. Rinse in distilled H20.
  3. Fix in formol calcium 15 minutes.
  4. Counterstain in haematoxylin 15 seconds.
  5. Wash and Blue.
  6. Dehydrate, clear . Mount sections in DPX

RESULTS

Brown reaction product indicates cytochrome oxidase activity

Note: DAB - Carcinogenic - HANDLE ACCORDING TO COSHH PROTOCOL.

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