Tuesday, 6-8-99: Took reading for experiment 2.  As I suspected the absorbance is higher in the low number of cells.  This can  be explained by the following:  MTS is not taken up as easily by cells that are higher in competition because their metabolism is lower than cells which still have adequate medium.  Solution:  More medium?  Less cells?

Wednesday, 6-9-99:  Planned experiment 3, plates and amount of medium per well etc.

Thursday, 6-10-99:  New experiment-referred to as exp. 3 started today.  New rats came in Tuesday and were allowed 48 hours to rest.
    The extraction of macrophages from the lung needed for this experiment involves lavaging the lungs with ph 7.4 PBS.  The first surgery went as planned but the rat was abnormally small and he died as soon as the pentabarbitol (a .8cc dose) was given.  Extraction of the lungs showed heavy blood clots in the lungs.  A second rat was obtained and 4.8 million macs were obtained specifically.
The plate plan for this experiment is as follows:

30,000 cells/ well   6 replicates. 1ng/ml GMCSF in first column only.  200uL/well.
 

Once the cells were plated, I incubated them for 1 hour.  I plated 9 plates total.  I took a DAY 0 reading on one of the plates.  I then saved this plate for a 24 hour reading.  The results will be posted at the end of this experiment.

Friday, 6-11-99: DAY 1 of experiment three yielded satisfactory results at the 60 minute reading.  After taking that reading, I also popped all the bubbles in the plate (since it is not sterile anymore) and took another reading with similar results.