HI, I have a qestion, I always got less than 105 cell on...

if you can detect IFNgamma and IL-4, they should differentiate into Th1 and Th2. For less cell No. problem, one possibility is that you lose many cells during staining and washing. You can count cells before you collect and stain them, if they are not less than 105, you need to think about your staining process. If before staining you have small number of cells, do not use CD3 after transferring cells to new plate, because anti-CD3 will cause cell death, that's why anti-CD3 is plate-bound.

th1,th17,treg , harvest three wells in each group，thanks！

1.5x105/well,

May I know which population you detect is less than 105? What's the number of cells when you start this experiment?