I'm trying to measure cell size using flow. It's simple enough but I find when I use a hypotonic solution, my cells appear to be shrinking!! I grow my cells in submerged culture and split them from the flask. Once I get a cell pellet I resuspend this in a bath solution of 140mM NaCl, 2.5mM KCl, 1.2mM CaCl2, 0.5mM MgCl2, 5mM Glucose and 10mM HEPES. The hypotonic contains 75mM NaCl.
Any ideas what's going wrong or any suggestions as to how to do it differently?