Sooo...I have been using counting beads to quantify the concentration of RBCs in a mouse. However, when I count 1000 beads/sample for example, half of my bead population binds to my RBCs. Initially, I was gating out beads from my control "just bead" sample on Forward/Side scatter. Therefore, in my calculation I was using 500beads instead of 1000. When I include 1000 beads in my calculation, my concentration of RBCs, halves which makes sense because my bead volume is increased but RBC number is staying the same. However, my supervisor thinks that if I use 1000 beads in my calculation my conc. should double. He explained it, it made sense, but now I go back to my workings it doesn't...anyone able to explain a) have your counting beads ever bound to RBCs and re-explain what my supervisor means....hopefully I make sense!