C2C12 Muscle Cell Imaging using Quantum Dots
Almodovar-Cruz, Janice M.
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C2C12 cells are a mouse myoblast cell line that is differentiates rapidly, forming contractile myotubes and producing characteristic muscle proteins. This research is about imaging these cells to track their motion under stimulus chemically or electrically. Also, we would like to perform experiments to track how fast DiI is digested by the cells. The method is to use florescent quantum dots and dyes attached to cytoskeletal markers as proteins to know which produce more fluorescence in the Deltavision microscope. The techniques used included: growing cells in a 75 mL plate with a dilution 1:15 waiting approximately 70% of confluent to split them, counting cells using an hematocitometer, using fibronectin and collagen coated to add the DiI and using quantum dot conjugated with Î±-5 IgG. These protocols use the TRITC microscope channel because the red filter which excites the DiI fluorescence and the antibodies conjugated with the quantum dots. Our DiI experiments demonstrate that the fluorescence decrease when the time passes. QDs experiments proved when it’s conjugated with Î±-11 and binds to the cell not lose fluorescence as normal dyes. We assume that the quantum dot has a long fluorescence lifetime and photostability. However, we must continue working with the QD conjugated with Î±-5 because so far not produce fluorescence and has not been linked to cells. For future experiments we will go to work with quantum dots conjugated with secondary antibodies, but first we will have to do an antibody reduction using SDS-PAGE.