WEEK TWO PAGE FOR SUMMER 99 EXPERIMENTS AT THE UNIV. OF MICHIGAN

Monday 5-17: Looked like not much work to do today. Set up meeting with Go-shau for protocols necessary for ICAM experiment. Looked at experiment under ELISA plate using the same concentration of MTS/PMS mixture. I waited for 60 mins and looked at the plate at 490 nm. The results were consistent with the hypothesis that more GM-CSF corresponds to more macrophage proliferation. The higher concentration of GM-CSF didn't have as much of an impact on the number at the lower concetrations resulting in a line that seemed to be leveling off at higher concentrations. See graph below:

Given these results, we concluded that the MTS assay will work for our later experiments. Donna was doing some flow-cytometry of epithelium cells labelled with PI. This stain labels the DNA allowing us to see what stage the cells are in. The experiment consists of cells deprived of serum and exposed to certain cytokines. I sat in with her and helped her perform this procedure. I am very thankful to her for explaining the procedures for flow cytometry. Thanks Donna!

Wednesday 5-19: Helped Donna with her flow cytometry today. Her experiment involves cells of the T2I cell line which is a lung epithelium cell line. Conditions which she exposes them to are combinations of Serum and GMCSF and the flow cytometry deals with finding out what phase of the cell cycle most of the cells are in after two days of experimental conditions.

Thursday 5-20: Planning to extract and isolate type to lung cells from a rat tomorrow so had to get all the solutions ready. Long day. Everything must be ready because the procedure itself is very long. Got the protocol and adapted it to my specifications. I'll put it on here after I actually do the experiment so any changes can be recorded.

Friday 5-21: Couldn't do the above planned experiment today. Got details about another one of my experiments in which I need the media from the growth of T2I cells after four days. So I had to expand the cell line. Monday is the rat sacrifice.

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