This week we completed the
experimental procedure I discussed in my previous post, and during this process
I had the novel experience of working with the flow cytometry machine. Flow
cytometry is a procedure by which cells, suspended in a fluid, are pumped one
at a time through an extremely narrow tube. As the cells flow through, they are
lit by a laser, and a receptor on the other side records the degree of
fluorescence that the cells exhibit. We used this machine to test for yellow
and red fluorescence in order to quantify the amount of E2F1 and DNA in the
cells, respectively.
Unfortunately, we did not grow
enough cells, and so we are repeating the 5 day cell phase experiment next
week. We had an adequate amount of cells grown in 10% BGS media, because that
quantity of BGS allows cells to go into proliferation. However, we also grew
cells in starvation media, which contains much less BGS. Placing cells in starvation
media pushes the cells into quiescence, preventing them from dividing. We had
hoped that by placing the cells in starvation media the transient color would
be present to a higher degree at day three, but forgot to account for the fact
that the cells would not divide when we seeded the plates.
