For the foreseeable future this would be be my last week of my URSS project. This week I completed one last passage and then transfected my construct into both the HeLa and Cx32 stable cell lines. Once the transfection was completed, and all measures had been taken to maintain the cells health, the setup for data collection was complete.
The protocol I would be undertaking for data collection is Dye loading. This involves flooding the outside of the cell with dye and then placing the cell under different conditions. In my case this would be 35mmHg aCSF and 70mmHg aCSF. In Wild-type Cx32 the 70mmHg aCSF would induce opening in the Cx32, allowing the dye into the cell, illuminating it. A video of the Rig I used can be found below.
As the project is drawing to an end I have had time to reflect upon it. This project has benefited me in a multitude of ways. Not only have I got experience in molecular techniques, cell culture protocols and also in dye loading, but I also learned lessons about how to approach research. Failure and mistakes are part of research, and I have learnt to embrace them and use them as a learning experience. This project has proved to me that research is my preferred career choice. The fact that I could experience this researching CMTX is a privilege. The prospect of this research being used to further the treatment and knowledge of this disease is exciting to say the least, and is a major driving factor that motivates me to do research.
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