The rats were anesthetized by spontaneous sevoflurane inhalation and were sacrificed by decapitation. The entire triceps of the lower leg were removed, fixed in 10% formalin for 1 month, and macerated with 3% potassium hydroxide solution containing 0.2% hydrogen peroxide for 3 weeks. The macerated muscles were subsequently decalcified in Sihler's solution I (one part of glacial acetic acid, two parts of glycerine, and 12 parts of 1% aqueous chloral hydrate) for 3 weeks, stained with Sihler's solution II (one part of Ehrlich's haematoxylin, two parts of glycerine, and 12 parts of 1% aqueous chloral hydrate) for 4 weeks, destained in Sihler's solution I for 3–24 h, neutralized in 0.05% lithium carbonate solution for 2 h, and then hyalinised using glycerine gradients [40, 60, 80, and 100% (1 week in each concentration)]. The location of the intramuscular nerve-branch-dense region was observed under an X-ray view box and then photographed. The relative location of the INDR and its center along the muscle belly length in percentile measures was determined using a micrometer.
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