Day 5 - A Day in the Life of a Budding Scientist

For the first day of this second week, I made my way up to the lab once again. I started off my morning abruptly by continuing the lab work that the doctor I've been shadowing has been working on since last Monday. She started by plating 6 wells, and cleaned out the prior cell flasks that were used with PBS (Phosphate-buffered saline), so that the serum from the media that was used last Friday for the cell cultures would go away. If the serum from the media wasn't taken away, it wouldn't allow the cell bonds to break like we needed them to. She then added Trypsin to the cell flasks so that the cell bonds could then break. Then, we spun the flasks in the centrifuge, so that all of the cells could fall to the bottom of the flask, and form into a pellet at the bottom
The cells were then resuspended in the media, checked for their cell count, and finally plated into the wells with a certain amount of media (dependent on their cell count).

Then, in the afternoon we started RNA Isolation of the CD73 gene in mice that the lab already started experimenting on. There were many experiments going on at once, so there was some multitasking involved as well. As well as the RNA Isolation, there was also the preparation of Electrophoresis, which where we just prepared the gel needed for the rest of the experiment tomorrow. For the RNA Isolation of CD73, it was somewhat similar to the cell culture in the morning, because after adding the lysis buffer to the mice's RNA, they are also spun in the centrifuge to get a cell pellet at the bottom. The last experiment that was also going on was the use of the leftover lysis buffer to create some Lysate, which needed to be made by putting the lysis buffer in ice, and moving it around in the wells for around 15 minutes. The RNA Isolation had to sit for 15 minutes as well.

It seems to be as though the experiments that I am involved in in the lab are seeming to get more and more interesting, as I'm also finally starting to fully understand the specific aspects of cancer research, immunotherapy and immunology as a whole. After heading home, I also think this was another great start to the second week of this J-Term.

Comments

Post a Comment

Popular posts from this blog

Day 8 - Adenosine All Over

Today, I had gone up to the lab I've been working in for the past two weeks. I started to read another grant that was about the role of adenosine in the function of immune cells, and how it's immunosuppresent. Later, I went to see the doctor I had been shadowing working, as well as operating on mice livers to see the role of low blood flow. That is one of the many things that adenosine receptors are needed for as well. I also helped grow macrophages, and did cell culture with them by adding more media to the already plated cell cultures that were done a few days prior. What I also saw the doctor do is start staining the cells from the liver, so that he could see how many T cells and macrophages there would be left after a liver injury, in the mice. This was a very slow day in the lab, as there wasn't very much to do today. Afterwards, I headed home, concluding my second to last day at Memorial Hermann.
Read more

Day 4 - The Start of Experimentation

For the last day of the first week of my J-Term, I had finally started to work in the lab itself. I arrived at Memorial Hermann in the morning, and made my way to the same lab once again. I was then greeted by the doctor that I would be shadowing and working with today. We both walked over to the cell culture room, so that she could explain to me the ongoing experiment that she was working on. She was working on cell cultures of course, and specifically, how CD73 plays a role in turning Adenosine into ATP, and how silencing that same CD73 changes the process as a whole. After a brief explanation of what we would be doing for that morning, for the remainder of this morning I watched her work on the controls that she needed for this experiment, as she already finished with the actual cell cultures a few days before, on Monday. She also explained what she was using for the controls, like the media, lipofectamine, etc. After she finished, we let them sit for 5 hours, and I went off to l
Read more

Day 6 - Of Mice and Men

I headed up to the lab in Memorial Hermann once again, starting new lab work that I wasn't familiar with yet. I first went to see the mice that the doctors had been working with so far, and waited out after they had to collect the red blood cells and bone marrow derived macrophages from the mice. They took the long bones of the mouse, cleaned off all the muscle, and flushed out all the bone marrow of all of its cells. Then, the docor filtered the cells to get rid of the remaining connective tissue that he didn't need, and spun them down in the centrifuge. He then resuspended them in ACK Buffer for five minutes, to get make the red blood cells burst, as he only needed to the bone marrow derived macrophages for these cell cultures. They were then spun down again to get rid of the red blood cells, and just have bone marrow pellets at the bottom of the flasks. The remaining bone marrow macrophages were resuspended in the media, and plated in the cell culture flasks with 20mls of t
Read more