We undertook a three-dimensional reconstruction of intramuscular collagen networks of bovine muscle using an immunohistochemical/confocal laser-scanning microscopic method. By immunohistochemical staining, type I and III collagens were observed mainly in the perimysium, while type IV collagen was observed in the endomysium. On the other hand, type V and VI collagens were observed in both the perimysium and endomysium. By confocal laser-scanning microscopy, the collagen observed in the perimysium was three-dimensionally reconstructed as plate-shaped layers whereas the collagen observed in the endomysium surrounded myofibers. The three-dimensionally reconstructed observations using immunohistochemical/confocal laser-scanning microscopic method is useful for investigating collagen networks in muscle.