Abstract
A confocal laser scanning microscope was successfully
used to visualize the T-system in skeletal
muscle fibers of the frog skeletal muscle stained with
the Golgi silver method. The muscle tissues were
selectively stained for the T-system and prepared
for electron microscopic observations. The staining
pattern varied among muscle fibers, but the fibers
selectively stained for the T-system could be readily
identified in reflected light by their yellowish color
among black-colored unstained fibers on the
trimmed face of an Araldite-embedded block under
a dissecting microscope. Under a confocal laser
scanning microscope with a reflection filter, the
stained fibers showed characteristic networks of the
T-system at the level of the Z-disc, providing sharp
and high-contrast images in optical sections of thick
slices. Such images resembled those obtained with a
transmission electron microscope in thick sections
cut from the same blocks. Sharp optical section
images were obtained at depths up to ～10 μm from
the specimen surface. The three-dimensional distribution
of the T-tubules was examined in stereo-pair
images reconstructed from serial optical sections. In
such stereo images, one could follow the plane of the
network of the T-system, often recognizing a helicoidal
pattern of the plane of the T-system.
