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A new technique, which clearly distinguishes fibre types in fixed muscle tissue
Author(s) -
Behan W. M. H.,
Cossar D. W.,
Madden H. A.,
McKay I. C.
Publication year - 2002
Publication title -
neuropathology and applied neurobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.538
H-Index - 95
eISSN - 1365-2990
pISSN - 0305-1846
DOI - 10.1046/j.1365-2990.2002.39286_58.x
Subject(s) - gene isoform , myosin , myofibril , immunohistochemistry , wax , frozen section procedure , muscle tissue , atpase , skeletal muscle , chemistry , monoclonal antibody , anatomy , biology , pathology , biomedical engineering , biochemistry , antibody , medicine , enzyme , immunology , gene
Aims: A method of distinguishing between type 1 and 2 skeletal muscle fibres in wax‐embedded tissue is needed. The ATPase method is the basis for fibre identification on frozen tissue and a new method should not give significantly different results. Isoforms of myosin and myofibrillar ATPase are known to correlate. Materials and methods: We devised an immunohistochemical double‐labelling (IHC) protocol using monoclonal antibodies to fast and slow myosin. We compared results in the two methods by morphometric analysis of frozen muscle and then applied the method to paraffin‐embedded tissue. Results: On frozen sections there were no significant differences ( P = 0.57) in the percentages of type 1 (46% IHC method vs. 48% ATPase) or type 2 fibres (54% vs. 52%) and 2a and 2b subtypes were distinguished easily. Cross‐sectional area (in µm 2 ), diameter (µm) and form factor were all similar. Various diagnostic samples of wax embedded tissue were then examined. These gave excellent results with clear colour contrast: type 1 fibres, black and type 2 fibres, red (see examples). Conclusion: An IHC method based on the fast and slow isoforms of myosin gives similar results to the ATPase method while providing an important advantage in its applicability to wax‐embedded muscle.