My third week in the Change lab has been amazing, and I learned a lot! My week started off pretty standard. I finished extra data entry from last week and worked with the days FORCE participants.
After learning that I passed my biosaftey training course I got to work in the wet lab for the first time! I started by shadowing an undergraduate student from CU in the lab while he prepped blood for testing. The blood we worked with was from the OASIS study (which is one I haven’t worked on before). First we centrifuged the blood in order to separate out the plasma. We also centrifuged using CPT tubes (cell preparation tubes) to separate the blood. CPT tubes have a filter that, when spun, lets the regular blood go through it while trapping plasma, certain cell types, and proteins. After this we pipetted a whole blood sample(not centrifuged) into tubes and stuck them in the freezer, which is kept at -80 degrees Celsius! After this was done we took more blood samples and added a PBS (phosphate buffered saline) solution. When PBS is added and centrifuged it helps to wash the desired cells to make sure there isn’t anything that could disrupt the test. Next, we plated some of the blood sample. This consisted of adding 3 mL of blood only containing PBS to two of the basins to use as a control. There is also 3 basins filled with blood containing LPS (lipopolysaccharide) which stimulates inflammation and infection. With the LPS the blood will be tested on its ability to create an immune response to the inflammation and infection. After all five of the basins are filled they are put in an incubator for 24 hours. In addition to prepping the blood for LPS testing, we also prepped for PBMC(peripheral blood mono nuclear cells) testing. PBMC’s are part of the immune system, and we prepped them by isolating them and then freezing them. This was a really interesting day. I was handed a lot of information at one time, but I learned a ton!
I was able to work in the wet lab again later in the week! This time I worked under someone else with blood from the COSMIC study. To begin we took one of the plated blood samples that had been in the incubator for 24 hours and worked with it. We separated the controls and the ones with LPS and did another PBS wash to ensure that the blood samples only had the desired cells for testing. The PBS separates out clotting factors, plasma, and platelets that might get stuck to the cannabinoids and cytokines, which are what is being tested. Cannabinoids are components of cannabis that are tested for the COSMIC study. Cytokines are just types of immune cells that the study screens for. After this I got to extract some plasma myself! I had to wear gloves and spray my gloves with ethanol before I could start. All of the blood extracting has to be done in a biosaftey cabinet where everything has been previously sprayed with ethanol. To extract the plasma a pipette is set to how every many mL is required for the test, then a clean tip is put on and the plasma is extracted. The tip then has to be disposed of in an autoclave bag for safety concerns. I loved working with the blood samples! I learned so much in just two days, and it has definitely been my favorite part!
On Wednesday this week I didn’t have to work, so I went to Eldorado spring with my host family! We went a short hike. We saw tons of climbers and other hikers there! It was really cool, and we would have kept going but it started to thunderstorm!
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