Blog 3

Since the last update, I have been able to design the protocols for my control experiments. This includes analyzing VASP location in wild-type, untreated macrophages, macrophages with small beads, macrophages with the addition of factors needed for activation, and macrophages with beads and the factors needed for activation. Each of these controls will need to be repeated three times to obtain results that can be analyzed using statistics. I will also be working on creating Mycobacterium bovis-BCG that is fluorescently labeled with GFP (green fluorescent protein). Once this is generated and my controls have been completed and analyzed, I will be able to infect the macrophages with the bacteria and look at the location of VASP in the macrophage. These experiments will require the use of primary and secondary antibodies for immunofluorescent examination on the fluorescent microscope.
One challenge I am currently facing is due to the computer connected to the microscope crashing. This has caused me to delay my control experiments because the cells cannot be analyzed without the fluorescent microscope. Successes of this project have included correct culturing of the macrophages and actively growing M. bovis-BCG. So far this project has strengthened my ability to design protocols. It has also helped me to learn more about literature searches and how to solve setbacks. I am looking forward to starting my control experiments as soon as the computer is repaired.

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