Week 3 – Getting the facts from FACS analysis

Mar 11, 2019

In my previous post, I mentioned that in every trial, there are two stages. In this week’s post, I will primarily be talking about the first stage, which is treatment and FACS analysis. At least 12 hours after treatment, I check the cells under the microscope first to see if the drugs actually had any effect. Then, I collect them and insert them into a flow cytometer machine, which spits out different kinds of FACS data. The data I am interested in is the live-dead data and the cell-cycle data. The live-dead data shows levels of live and dead cells, and the cell-cycle data shows the part of the cell cycle at which the live cells were at.

FACS Analysis stands for Fluorescence-Activated Cell Sorting, and it is used to profile a large population of cells in a liquid media. In a flow cytometer, cells are passed through a narrow channel, and light is used to illuminate the cells in that channel. Sensors then detect where the light is reflected off the cells, and the cells are plotted on a graph. Raw data shows each sample’s graph with the x-axis of forward scattered light, which means the light is reflected in the same direction as it was emitted, and the y-axis of side-scattered light, which means the light is reflected in different directions.

In the picture below, you see sample graphs of my data. This is just one trial, and only 6 of the 24 conditions. As you can see, we have 2 levels, Apo (apoptosis) and Live (…which means live). In the first graph for example, 39.9% of the cells are dead because of apoptosis, and 59.3% are still alive.

I will be analyzing several more graphs like this to determine what is the best treatment dosage. From there, I will choose 10 conditions to test for western blot.

One Reply to “Week 3 – Getting the facts from FACS analysis”

  1. Sahithi P. says:

    Are you going to run the test multiple times, or is one run enough to determine the treatment dosage?

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