Figure 3

Immunohistochemistry of cell pellets of myotube cultures. 3,3^′ diaminobenzidintetrahydrochlorid (= DAB) stainings of canine myotubes, Primary antibody: mouse anti-Desmin, secondary antibody: goat-anti-mouse-biotinylated, A, B: (A 100-fold and B 400-fold mag.) primary culture prior to freezing, approximately 70% to 80% of the cells are positive for the muscle-specific intermediate filament desmin. C, D (C 100-fold and D 400-fold mag.) showing a cell culture pellet after freezing and recultivation. Estimated value of desmin positive cells is 40%. Structural integrity of myotubes is intact, but morphology has been lost due to pellet preparation. E: ascites of non-immunised BALB/cJ mice in a dilution of 1:1000 as negative control (100-fold mag.). F: normal canine skeletal muscle tissue as positive control, few vimentin-expressing cells (100-fold mag.). Bar= 100 μm. mag. = magnification.