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Application of the fractionator and vertical slices to estimate total capillary length in skeletal muscle

  • EMILIO ARTACHO-PÉRULA (a1), RAFAEL ROLDÁN-VILLALOBOS (a1) and LUIS M. CRUZ-ORIVE (a2)

Abstract

A new stereological method is proposed which combines vertical slice projections with the fractionator to estimate the total capillary length in a skeletal muscle. The method was demonstrated on the soleus muscle of a Wistar rat. The implementation required capillary highlighting, tissue sampling, and data acquisition in the form of intersection counts between capillary projections and cycloid test lines. The capillaries were demonstrated using vascular perfusion (with gelatine) of the hind leg of the rat. The sampling procedure followed the fractionator design, namely a multistage systematic sampling design with a known sampling fraction at each stage. To make the design unbiased, vertical slices were used; for efficiency, the vertical axis was chosen parallel to the main axis of the muscle. As prescribed to avoid bias, the cycloid test lines were superimposed on the slice projections, viewed under the light microscope, with their minor axes normal to the vertical axis. The estimation precision was compared for different sampling and subsampling fractions. The proposed method was globally highly efficient, unbiased, and easy to implement.

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Corresponding author

Correspondence to Dr E. Artacho-Pérula, Department of Morphological Sciences (Section of Histology), School of Medicine, University of Córdoba. Avda. Menéndez Pidal, s/n. E-14071 Córdoba (Spain). Tel: +34 957 218261; fax: +34 957 218246; e-mail: cm1arpee@lucano.uco.es

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Application of the fractionator and vertical slices to estimate total capillary length in skeletal muscle

  • EMILIO ARTACHO-PÉRULA (a1), RAFAEL ROLDÁN-VILLALOBOS (a1) and LUIS M. CRUZ-ORIVE (a2)

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