Nuclear microscopy of normal and necrotic skeletal muscle fibers: an intracellular elemental microanalysis

The in situ total elemental composition and elemental concentrations present in mouse soleus (type I) and gastrocnemius (type IIA) muscle fibers were analyzed by using nuclear microscopy (NM). Elemental changes in necrotic fibers, induced by intramuscular injection with snake venom (Pseudechis australis), were also studied 3 h post-injection. Nuclear microscopy is a new method based on nuclear technology that utilizes the interaction between a million-electron-volt nuclear particle beam and the muscle sample (in the case of the present study). Elemental analysis was done at the parts per million (ppm) level of sensitivity on unfixed, rapidly frozen and unstained single fast- and slow-twitch muscle fibers with imaging capabilities of micron spatial resolution and in multi-elemental mode. In total, 12 different intracellular elements were mapped, co-localized and analyzed in single normal and necrotic skeletal muscle fibers from mice. Elements such as potassium, sulfur, phosphorus, chlorine and sodium were found in concentrations from 1000 to 18,000 ppm. Unlike conventional electron-probe X-ray microanalysis, NM also detected and analyzed the trace elements such as magnesium, calcium, iron and zinc that were found in concentrations of 50 to 1000 ppm. Other elements--copper, manganese and rubidium--were also detected in concentrations of less than 50 ppm. The trace elements calcium, iron and zinc were more abundant in the soleus than the gastrocnemius (the level of iron was statistically significant). Calcium, sodium and chlorine were significantly elevated in venom-induced necrotic soleus muscle fibers.