Histotechnology Technical methods

Connective Tissue Stain for Muscle Biopsy

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


Staining solution

  • Chromotrope 2R 0.6g
  • Fast green FCF 0.3g
  • Phosphotungstic acid 0.6g
  • Glacial acetic acid 1.0ml
  • Distilled water 100ml
  • Adjust to pH 3.4


  1. Stain nuclei well (one minute) with haematoxylin and wash well in water.
  2. Stain in staining solution for approximately 30 seconds-1 mintue.
  3. Rinse in 0.2% acetic acid.
  4. Dehydrate, clear . Mount sections in DPX


Nemaline rods, ragged red fibres . Red on a blue/green background.

This method is virtually only used on muscle biopsies. The staining solution deteriorates with age, so if only used rarely it should be made up fresh each time.

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