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Flow Cytometry

After the cell lines have been established from single colonies of the green fluorescent cells that were collected through cell sorting, the cells are put into the flow cytometer. The flow cytometer confirms that the 4T1 cells have been successfully modified and that the plasmid was correctly transfected. The flow cytometer works by chemically measuring the proteins being produced by the 4T1 cells. It allows researchers to visualize and verify the presence of their target proteins. Flow cytometry is doubly useful in studying and identifying mixed populations of cells since the flow cytometer can identify up to 16 colors of fluorescently tagged cells, while the cell sorter can only identify up to 8. The flow cytometer is also faster and more accurate than the cell sorter. Using this machine allows verification that the cells have been modified in the most secure, sure way possible. However, there is a distinct difference in the cell sorter and flow cytometer. The cell sorter is able to separate and analyze cells while still keeping them alive; on the other hand, the flow cytometer kills the cells. Thus, the cell sorter is used first so cell lines can be established, even though the flow cytometer is more efficient.

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