I'm planning an experiment where I can access intracellular cytokines in a specific subregion of the brain in mice. However, this brain region is quite small, maybe 50,000 cells per animal. I know I will need to pool mice but how many would I need to pool? Can I use 500,000 cells? Pooling more than 10 mice wouldn't be feasible.
Thank you!
Hello Kallon Crowther
Yes, you may use 500,000 cells. For your experiment you will need cells in the range of 10^5 to 10^6. It is better to be prepared for a higher cell number in case of any error during the experiment.
Best.
If possible if you know the frequency of your positive population that can also help determine how many cells to add per well. Rare population of cells would be harder/impossible to see if you add too few cells. The numbers indicated by Malcom are good, but in general its good to have an idea (if possible) of your % positive population (out of total live cells)
I agree with Aaron Dhanda you need to perform flow on single dissociated region to determine what is your positive population Then you can estimate how many mice you need and how feasible this experiment.