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

Published online by Cambridge University Press:  01 October 1999

EMILIO ARTACHO-PÉRULA
Affiliation:
Department of Morphological Sciences (Section of Histology), School of Medicine, University of Córdoba, Spain
RAFAEL ROLDÁN-VILLALOBOS
Affiliation:
Department of Morphological Sciences (Section of Histology), School of Medicine, University of Córdoba, Spain
LUIS M. CRUZ-ORIVE
Affiliation:
Department of Mathematics, Statistics and Computation, Faculty of Sciences, University of Cantabria, Santander, Spain
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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.

Type
Research Article
Copyright
© Anatomical Society of Great Britain and Ireland 1999

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