Hello Harbour Friends, today was VERY exciting. We are going to again delve into the world of biology and cancer research from my times at NIH!
Today was very exciting. I was able to check on my cells from last week, which to my surprise, were clean! They grew perfectly with no sight of bacteria, fungi, or other contaminants. I guess you could say I’m getting the hang of this. I was able to take last week’s cells and further multiply them by transferring cells into more growth media with little to no help from Mr. Choh. From now on I will refer to “Dr. Choh as “Mr. Choh”. I walked in today and asked for Dr. Choh, he shook my hand and humbly said, “I am not a Dr.”
The steps are as follows:
Aspirate solution (suction it off)
Add phosphate buffer solution (also known as PBS)
Close flask and rock solution around to balance the pH
Aspirate the PBS solution
Add 1 ml of trypsin and rock back and forth, let it sit for a minute then smack the side of the flask
Add 10 ml of growth media
Take five ml of solution and transfer
Add fifteen ml of growth media to original flask
I did this about 4 or so times for the RD and RH-30 cancer cell lineage. I added the 5 ml samples to test tubes that I will use later. After this, I went to lunch. They have quite a nice cafeteria area, with a coffee shop as well as a gift shop. The gift shop has nothing really of interest though. I should get a gift for my friend just to mess with him; he doesn’t always like gifts, especially if they are weird or useless .
After this it was back to the lab. I had to first count how many cells I had grown. They use these slides that have 2 little wells, one for inserting the cell culture, and another for the air to escape, after this is done the slide locks both wells. I was then brought to a very big machine, about the size of a desktop computer tower. This machine had a slot that you inserted the slide into. The machine was called cellometer. The machine was connected to a Macintosh computer via USB. You open a program called Cellometer and you hit display image. The microscope inside the machine takes a small square picture of your cells. You then enter the cell type and hit count cells. The machine counts the cells and does the calculations for how many cells you have per ml of fluid. We analyzed the data and plated the cells. In plating the cells you have to make SURE that you have only 20,000 cells per well. The plate is made up of 96 wells that you can divide and label into different sections. I had to grab only a few µl’s (microliters) of the cells. We divided the plate into two halves, each half being a different lineage of cells. Then we had 2 sections of 4 rows on each half. Rows 1- 4 were used to give a control, low dosage of cancer drug treatment, moderate dose, then the highest dose. Rows 5-8 were used for control, low dose of antibody, moderate dose, and then high. We will see how quickly these cells grow in next week’s post.
Overall, I had a wonderful time! Again, I look forward to any feedback on this week’s blog! I look forward to talking about next week’s adventures. Thanks for reading, Aaron J.