Project Outline Part 2

To accomplish this, we followed the protocol from ThermoFisher titled, “ER-Tracker™ Dyes for Live-Cell Endoplasmic Reticulum Labeling”.  Initially we incubated the plate of cells at 370C for 30 minutes. Following the incubation period, the cells were again rinsed with PBS and given media without CO2. The resultant cells appeared to over-fluoresce in unexpected locations so we repeated the procedure. We decided to incubated for 20 minutes in the staining mixture to prevent the overfluorescense. The cells incubated for 20 minutes appeared to clearly mark parts of the ER. These distinct sections of the ER marked by the tracker did not seem to distinctly overlap with the portions of the ER marked by the plasmid. However, we can confidently say that the plasmid marked the ER, just not the same aspects as the ER tracker did.