Glomerular Airways Lab Report
OBJECTIVE
The objective of this lab was to determine the effects of two unknown drugs on the microtubule pathways in COS-7 cells through the process of immunostaining. METHOD
Before arrival, the COS-7 cells were grown in culture dishes submerged in a growth medium that contained bovine serum. Two coverslips covered in the COS-7 cells were provided. After washing the coverslip with Phosphate Buffer Saline (PBS), one coverslip was treated with the control solution, and the other was treated with the X drug. Then, in order to preserve the cell, methanol was added to both coverslips. Since methanol coagulates proteins, it was also used to permeabilize the cell membrane. Once the cells were fixed and permeabilized, blocking buffer was added to each coverslip to block the nonspecific antibodies in order to keep them from binding to everything in the cell. Then the primary mouse antibodies were added to the cells, and the secondary antibodies are added to bind to the primary antibodies. After these cells were given time to incubate with the second antibody, they were ready to be viewed. Using the mounting medium that contained DAPI, the coverslips were mounted on the slide for the microscope. …show more content…
In the drug treated cell, the centrosome is not visible, and there is a lack of microtubules in the space that the centrosome would presumably be. There is a surplus of microtubules lining the perimeter of the cell. There are also noticeably more microtubules in the overall cell. The nucleus (blue) looks more crisp and concentrated in the drug treated
The objective of this lab was to determine the effects of two unknown drugs on the microtubule pathways in COS-7 cells through the process of immunostaining. METHOD
Before arrival, the COS-7 cells were grown in culture dishes submerged in a growth medium that contained bovine serum. Two coverslips covered in the COS-7 cells were provided. After washing the coverslip with Phosphate Buffer Saline (PBS), one coverslip was treated with the control solution, and the other was treated with the X drug. Then, in order to preserve the cell, methanol was added to both coverslips. Since methanol coagulates proteins, it was also used to permeabilize the cell membrane. Once the cells were fixed and permeabilized, blocking buffer was added to each coverslip to block the nonspecific antibodies in order to keep them from binding to everything in the cell. Then the primary mouse antibodies were added to the cells, and the secondary antibodies are added to bind to the primary antibodies. After these cells were given time to incubate with the second antibody, they were ready to be viewed. Using the mounting medium that contained DAPI, the coverslips were mounted on the slide for the microscope. …show more content…
In the drug treated cell, the centrosome is not visible, and there is a lack of microtubules in the space that the centrosome would presumably be. There is a surplus of microtubules lining the perimeter of the cell. There are also noticeably more microtubules in the overall cell. The nucleus (blue) looks more crisp and concentrated in the drug treated