One thing I learned that day- learning to use excel is something you need to do... no matter what. It is extremely useful and actually kind of fun (once you get the hang of it). Alas, this time I did not use excel. Instead, I did everything the old school way (which is surprising since I belong to the tech generation)- with paper and pen (and a calculator, but that's beside the point). After about 30 to 45 minutes of crunching numbers and filling out the excel sheet I emailed Dr. Lacombe my final file, with the data, the graph, everything.
Before comparing our analysis, we went to the culture room. in the cell culture, we changed the medium of the PC3 clonogenic assay, PC3 cell line flasks, and 22Rv1 cell line flasks. We also passed the DU145 cell line into new flasks and froze 1 mL of each flask.
Once that was completed, Dr. Lacombe and I reviewed the clonogenic assay. Here is a my graph and Dr. Lacombe's graph.
The discrepancies between our graphs is due to errors in counting the colonies. When counting, I either counted colonies that had less than 50 cells or counted colonies that were close together as one when they should have been counted separately.
The expected result were:
1. The non-irradiated cell line would be the least radioresistant- meaning the most number of cells would have died in this cell line
2. The cells that received the fractionated dose (5x2Gy) should have been the most radioresistant. Each time the cells were irradiated a portion of the cells survived. These cells exhibited radioresistance meaning each time they replicated the daughter cells would inherit this radioresistance. In the end, most of the cells on this plate should have been radioresistant and survived.
The analyzed results did not match this expectation. Instead, the non-irradiated cells turned out to be the most radioresistant and the cells that received the fractionated dose were the most radiosensitive.
Interesting. So, referring to the difference between your analyzed results and expected results, is it a significant difference? What does the difference indicate to you? What does it mean in terms of your project and your goal?
ReplyDeleteGreat question. To see if the results are significant and to make sure that this is the actual trend we will have to make another clonogenic assay. This will happen in the next week or so. Hopefully I'll be able to answer your question in greater detail then!
DeleteI love that you are sharing some of your data about your progress. It looks great! How will you be conducting your analyses? Will you use Excel, SPSS, SAS, or R for the actual statistics?
ReplyDeleteThank you and we will be using excel to conduct our analyses. Dr. Lacombe will be doing some of the analysis on his own as well.
Delete