Michael King - Week 7 - Loh Lab UCSF

Our cell cultures have finally started to grow healthily, and we were finally able to perform a few experiments on the cultures this week. The first experiment we performed was to treat five different cell lines with two different experimental drugs. After a 24 hour incubation, we determined the viability of each cell line with and without the drugs by first looking at the samples under a microscope, and later with the Vicell machine. After assessing viability, we performed a western blot to determine any changes in protein expression between treated and untreated cells. We also demoed a new gel reading machine from a local company, and the images turned out very well. My postdoc was pleased with the results from the experiment, as they clearly indicated there was one protein that would be induced when cells were susceptible to drug therapy.

I also set up my own experiment last week with just one cell line, but multiple types of chemotherapy. We will again perform a similar analysis of each sample with a western blot to look for protein markers. I am fairly independent in terms of taking care of the cells now, and I usually check viability everyday and look at them under the microscope to ensure they are not contaminated. I also have to prepare and change the media, and I have been coming in for around an hour each weekend to make sure the cells' media is replaced. All of this work has to be done under the hood to ensure sterility.

I also finally completed all of my mouse training, which involved me taking a three hour training class. I learned how to perform injections on mice and how to give them anesthesia. I also had to take the mandatory euthanasia training that involved severing the necks of mice. The mice are not the easiest to work with, and they do bite quite often.

We have been getting some very good results these past weeks, and everything is going smoothly. I only have two weeks left, and I have definitely learned a lot in the last six. My postdoc will not actually be around for the last week, so I will need to be even more independent in taking care of the cell cultures and performing western blots.