 |
 |
 |
|
|
|
STORAGE ASSOCIATED ARTEFACT IN EQUINE MUSCLE BIOPSY SAMPLES
Maile, C. A., Stanley, R. and Piercy, R.J.
Comparative Neuromuscular Diseases Laboratory, The Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, Hertfordshire, AL9 7TA.
Muscle biopsy in equine veterinary practice is increasingly used for investigating exercise-related myopathies and poor performance. Although traditional formalin fixation is sometimes used, for complete evaluation, fresh tissue is required that is frozen on arrival at a specialist laboratory.
Aims: To determine optimal storage conditions for fresh equine biopsy specimens submitted to laboratories and investigate the hypothesis that altered temperature, duration and hydration during storage influence severity of artefact.
Methods: A skeletal muscle sample was divided into 6 independent samples from each of 8 recently euthanased horses. Each sample was stored in a different way, designed to mimic possible conditions encountered in practice, including altered storage time, temperature and hydration status. All samples were then frozen in isopentane precooled in liquid nitrogen and sectioned and stained using haematoxylin and eosin and periodic acid schiff reagent. Stained sections were assessed and ranked on two occasions by two separate blinded observers.
Results: Weighted Kappa and Spearman’s rank correlation coefficients of inter- and intra-observer agreements were high. Both prolonged storage and increased hydration resulted in more storage-associated artefact.
Conclusions and Practical Significance: Results suggest that current recommendations for 24 hour storage of equine muscle may not be optimal: instead, preservation appeared best in samples stored at 0oC on dry (rather than damp) gauze. Although further study is required to determine whether this condition is suitable for all histochemical and immunohistochemical techniques, these results may have important implications for optimal shipping of equine muscle samples to laboratories.
BVetMed Final Year Elective Project presented by Charlotte Maile (cmaile@rvc.ac.uk) at the British Equine Veterinary Association (BEVA) Congress 2007
