This week has been a difficult week for everyone in my lab. Our coworker and good friend Mona Cornwell passed away over the weekend. Mona was a lab technician and histologist, so she was the one who was embedding all the tissue samples and making all the slides. Mona and my mentor Sue had worked together for over 30 years, and while Sue was busy doing her own work, Mona would take me and show me some stuff in the lab. I learned a lot from Mona and her and I had gotten pretty close because my mentor Sue often was working somewhere else, so Mona and I had spent a lot of time together. Mona was a friend to all and there wasn’t a mean bone in her body, she really made a huge impact on my experience at North Shore and I will never forget what she did for me. Needless to say it has been a quiet week in the lab and it has been tough for everyone.
This is the machine I use that reads the cells on my 96 well plates so we can have some sort of cell count that we can compare in different plates for different days.
For my project that I started last week, I continued and finished it and collected all my data. To collect my data I have to put my ninety-six well plates into a special machine that reads the amount of cells there are in each well, so then it transfers to a computer and I can put the data onto a spreadsheet. The results were not exactly what we wanted them to be, so we decided to do the experiment over again but add more variables to the cells. So now we not only have EGF and 0.5% media, we are also putting in the condition media, which is used food, from CAF cells and NPF cells. CAF (Cancer Associated Fibroblasts) cells are cancerous stromal cells and NPF (Normal Prostate Fibroblasts) cells are non cancerous prostate cells so we can compare with the CAF. Conditioned media is food that we put in the CAF and NPF flasks for 2 days and then we take out filter it then put it into the wells, and the theory here is that the cancerous stromal cells give off a signal to the cancerous epithelial cells that tells them to grow faster, and this signal is supposed to be in the media we took from the CAF flasks, while the NPF is not supposed to have this signal. So we are testing to see if the conditioned media from the CAF makes the cells grow more than the conditioned media from the NPF, and whether or not African American cells are affected differently. We also changed our slide layout a little bit so now we have five trials of each condition and we are only doing 5,000 cells per well because we thought that we saw better results in the 5,000 cell wells in the previous experiment.
This is a chart of my data from one of my days of data collection and it has the general layout of what we were expecting except we hoped to see that 0.5% (blue) was about half of 5% (green) and 1 ng/ml (yellow) and 100 ng/ml (red) were about ¾ of 10 ng/ml (orange) and 10 ng/ml was more than 5%. So that is why we decided to repeat the experiment, and since we had done it once, the second time has been much easier.
I got about halfway through this second project and next week I will finish collecting the data and give a presentation to my co-workers in the lab. Since I did the same experiment last week, this time it has gone much easier and I have done it with much more confidence, so I am sure that the data will be a little more reliable because I have more experience and I know exactly what I am doing, unlike the first time I did it.
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