Comparison of the Different Protocols for Fixation and Staining of Differentiated Skeletal Muscle Cells
LI Jing, LIU Lutao, GU Shuyu, CHEN Huaqun*
The effects, as well as the advantages and disadvantages of different fixation and chemical staining methods for differentiated skeletal muscle cells were compared. The possibility of the chemical staining applied in the analysis of the cells was explored. C2C12, the precursor skeletal muscle cell, was used in this study. After three days of myogenic differentiation, the cells were fixed with 4% PFA(paraformaldehyde), 100% methanol or Carnoy fixative for 5 min, 10 min, and 15 min, respectively. The best fixation protocol was thus determined. Then, the fixed cells were stained with the dye solutions of modified phenol fuchsin, Giemsa, or HE (hematoxylin-eosin). The results showed that among those protocols, the fixation with 4% PFA for 5 min or 10 min better preserved the morphology of the myotubes than the others, while the HE staining was the best one for the staining of differentiated myocytes including the formed myotubes and the unfused myocytes. However, immunofluorescence staining was better than all those three chemical staining protocols.
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