The immunofluorescence localization of beta-actin and phalloidin in C2C12 cells. The examination of the C2C12 cell line of mouse muscle cells under Olympus IX51 inverted fluorescence microscopy. The observed magnification was at 400X (A-E) with the use of CellSens imaging software. The cell line was cultured over a period of 12 days within a 10% FBS growth media. The cells were labelled with Beta-actin (green), Alexa Fluor 546 phalloidin (red), and DAPI (nucleus; blue). The days of fixation include prior to differentiation at myoblast stage (MB), Day 0, Day 5, Day 7, and negative control Day 12. The stages of differentiation within myogenesis include myoblasts to myotubes.

Beta-actin is present with sampling days of myoblast, Day 0 and day 5. Localization of beta-actin surrounds the nucleus within the myoblast and then slowly is distributed throughout the cell aiding is cell structure in differentiating myoblasts and growing myotubes on Day 0 and Day 4.

Preliminary evidence of beta-actin presents that it appears in the earlier stages of myogenesis as it depletes after days 4-5 once the majority of the cell group is differentiated into myotubes. This matched the theorized conclusion that beta-actin is primarily localized within the early stages of myogenesis.

Figure 2. The morphology and nucleation of C2C12 cells at different days of differentiation. The examination of the morphology of the C2C12 cell line under Olympus IX51 inverted microscopy and DIC (Differential interference contrast) microscopy. The observed magnification using the ocular lens at 40x and with the use of CellSens imaging software a DIC and DAPI overlay was finalized to the observing images. The cell line was cultured over a period of 10 days within a 10% FBS, DMEM, and 1% Penicillin-Streptomycin media. The nuclei were labelled with a DAPI stain (blue) to identify cell count and viability. The days of fixation include prior to differentiation at the myoblast stage (MB), Day 0, Day 4, Day 7, and the negative control at Day 6. The stages of differentiation within myogenesis include myoblasts to myotubes.

The morphological changes are visually seen as the myoblasts in MB and Day 0 are mono-nucleated and star-shaped. The myotubes are identified on days 4, 7, and 12 as multi-nucleated and rectangular-shaped

These are some other images of Fall semester 2021 of the sampling days variety between MB, Day 0, 4, and 7.

These images are not suited for comparison but are still used as a visual representation of beta-actin (green), phalloidin (red), and DAPI (blue).