Objectivity of two methods of differentiating fibre types and repeatability of measurements by application of the TEMA image analysis system.
Abstract: The objectivity of two of the most widely used methods for differentiation of fibre types, i.e. 1) the myosin ATP-ase method (Brooke and Kaiser, 1970a,b) and 2) the combined method, by which the myosin ATP-ase reaction is used to differentiate between fast and slow twitch fibres and NADH-tetrazolium reductase activity is used to identify the subgroups of fast twitch fibres (Ashmore and Doerr, 1970, Peter et al., 1972), was assessed in muscle samples from horses, calves and pigs. We also assessed the objectivity of the alpha-amylase-PAS preparation for the visualisation of capillaries (Andersen, 1975) in these species. For the purpose of reducing the time costs of histochemical analysis of muscle samples, we have developed an interactive image analysis system which is described. All analyses are performed on this system. In accordance with several other investigations, differences between the two methods of differentiating fibre types were found only for the relative distribution of the fast-twitch fibre subgroups (p < 0.02 and p < 0.05 for the relative number of type IIA and IIB, respectively) whereas when calculated in relative area, only the proportion of type IIA fibres differed significantly (p < 0.05). Significant interaction effects between method and person and method and species were found for the same traits. When methods were analysed separately, the combined method displayed a significant variance component of person for the distribution of type IIA fibres both in relative number and in relative area (p < 0.01 and p 87%), the impact of differences in pre-requisites (varied degrees of overlap between the fibre types) for performing the differentiation by the combined method raises a question of the reliability of this method. Apparently, no general rules for comparison of results of distribution of the two subgroups of fast twitch fibres by the two methods are applicable. The alpha-amylase-PAS method was found to be a fairly objective method to identify capillaries in muscles from horses, calves and pigs. However, as capillarity described in combination with other traits to give an indication of diffusion characteristics is significantly influenced by person, it is recommended that the same person perform all the analysis of a project. In addition to the methodological results in this study, we have shown that by application of the TEMA image analysis system, which is more rapid compared with the time-consuming traditional method for evaluation of histochemical preparations, analysis of samples from large-scale experiments are now possible.
Publication Date: 1998-06-06 PubMed ID: 9615191
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The article presents a study comparing the objectivity and efficiency of two methods used for differentiating muscle fibre types in animals. The study also uses the TEMA image analysis system to enhance the speed and precision of the measurements.
Objectivity of Muscle Fibre Differentiating Techniques
- The report examines the objectivity of two popularly used methods for distinguishing fibre types in the muscles of horses, calves and pigs. The subject methods are the Myosin ATP-ase method and the combined method (using Myosin ATP-ase and NADH-tetrazolium reductase reaction). Additionally, the alpha-amylase-PAS preparation, used for visualising capillaries, is also scrutinised for objectivity.
- The researchers found a minor difference in the relative distribution of fast-twitch fibre subgroups when using the two differentiation methods. The difference was significant only for Type IIA fibres when calculated based on relative area.
- There were notable interaction effects between the methodology used, person conducting the test and the species being examined.
- When both methods were tested separately, the combined method displayed a significantly varying component based on who was performing the test. This raised questions about the reliability of the combined method.
Use of the TEMA Image Analysis System
- The study also introduces an interactive image analysis system, aiming at reducing the time costs of histochemical analysis of muscle samples.
- The TEMA image analysis system was applied, resulting in increased efficiency in the traditional evaluation process. This method proved particularly useful for analysing samples from large-scale experiments.
Implications and Recommendations
- The researchers caution that no general rule can be made to compare results from the two methods for fast-twitch fibres.
- The alpha-amylase-PAS method is deemed fairly accurate for identifying capillaries in animal muscles. However, since capillarity, when combined with other traits, to show diffusion characteristics, is heavily influenced by the person conducting the test, it is advised that the same person perform all the tests within a project.
Cite This Article
APA
Henckel P, Ducro B, Oksbjerg N, Hassing L.
(1998).
Objectivity of two methods of differentiating fibre types and repeatability of measurements by application of the TEMA image analysis system.
Eur J Histochem, 42(1), 49-62.
Publication
Researcher Affiliations
- Danish Institute of Animal Science, Foulum Research Centre, Tjele, Denmark.
MeSH Terms
- Animals
- Capillaries
- Cattle
- Horses
- Image Processing, Computer-Assisted / methods
- Muscle Fibers, Skeletal / cytology
- Muscle, Skeletal / anatomy & histology
- Muscle, Skeletal / blood supply
- Reproducibility of Results
- Swine
Citations
This article has been cited 4 times.- Karen P, Števanec M, Smerdu V, Cvetko E, Kubínová L, Eržen I. Software for muscle fibre type classification and analysis. Eur J Histochem 2009 Jun 29;53(2):e11.
- Lee SH, Kim JM, Ryu YC, Ko KS. Effects of Morphological Characteristics of Muscle Fibers on Porcine Growth Performance and Pork Quality. Korean J Food Sci Anim Resour 2016 Oct 31;36(5):583-593.
- Meunier B, Picard B, Astruc T, Labas R. Development of image analysis tool for the classification of muscle fibre type using immunohistochemical staining. Histochem Cell Biol 2010 Sep;134(3):307-17.
- Karen P, Stevanec M, Smerdu V, Cvetko E, Kubínová L, Erzen I. Software for muscle fibre type classification and analysis. Eur J Histochem 2009 Apr-Jun;53(2):e11.
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