I have a problem with an experimental design that I hope someone might help me with. I also hope that I adequately explain the problem so that it is understood. We wish to look at the Calcium mobilization within endothelial cells using Indo-1. The stimulus we wish to use might be NK cells. The NK cells will be loaded with Fluo-3 since we are also interesed in the calcium dynamics of the stimulating cell as well. The problems with the lag between adding the cells and then continuing to acquire data are understood however, our problem is that we will have a nice base line for endothelial cells to which we can compare the resulting stimulation but we will not have such a base line for the NK cells since they will be added as the stimulus, making any data that we acquire regarding the Fluo-3 signal relatively uninterpretable. We have thought that we might run a baseline of the endothelial, remove the sample, continue along the same time line, put on a sample of NK cells, run a short baseline, return the endothelial cells to the run and then add the NK cells to them. This would give us a short endothelial and NK baseline and then we can collect the signals and potentially compare stimulation to baseline. Does this sound completely foolish? Would this be accepted as valid data? Is there any other way of accomplishing the same end result? We have considered reversing the order of adding the cells to see what was happening in separate experiments i.e. add NK to endothelial as one experiment and then the endothelial to the NK as a second experiment, but we would like to do both simultaneously if we could. Any comments and suggestions would be most appreciated. Haywood Pyle pyle@kfshrc.edu.sa
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:49:48 EST