Week Two: Experimental Preparation

Apr 02, 2019

This week I spent a lot of time mixing different solutions and stocks. On Monday I created a batch of PBS solution. PBS stands for Phosphate-buffered saline and it is used to maintain the pH of the cells. The cells that I incubate and use in the future would be suspended in PBS.

On Tuesday I began to grow fibroblast cells. Fibroblast cells are non cancerous cells and are the most common cells in the body. In the experiment, they will be used as the control. I grew them in a dish.

Wednesday and Thursday were very similar. On both days I created solutions of cells. I am currently using colon cells and they are very small and plentiful. I had to calculate the number of cells per milliliter and then dilute accordingly. I tried a ratio of 1:2, 1:3, and 1:4 of cells to PBS solution. I found that the ideal ratio of cells to work with was 1:3.

On Friday I began growing cells for my Monday assay. I used a 96 well plate and grew cells at densities of 1000, 2000, 4000, 6000, 8000, 10,000, 15,000, 20,000, 25,000, and 30,000. This was done with the colon cancer cells. I then placed the cells in the incubator because they require a minimum 48 hour growth period.

Overall, a lot of what I did this week was in preparation for the experiment next week. Next week I will be running cell assays on the cancer cells. Assay is a method by which to assess the cytotoxicity of a mater. It is used to calculate how many cells are dead or alive in the solution and whether or not that solution can be used for further experimentation.


One Reply to “Week Two: Experimental Preparation”

  1. William E. says:

    Hi Eva!
    It sounds like you had a fascinating week in the lab. Regarding the experiment that will be run next week, what do you predict the outcome will be? What percentage of the cells will be dead? What percentage will be alive? I’m also interested to find out whether or not the time the cells spend incubating in the PBS might affect the outcome during their experiment. I understand that PBS is used to maintain the cells ideal PH levels but might it have unintended side-effects that could tamper with the outcome of the experiment? I can’t wait to hear what results you find next week from your experiment!

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