-mix 10ul 0.4% Trypan Blue to 10ul cell samples, pipette to mix.

-load 10ul of the Trypan and cell mixture to either A or B chamber on the disposable slide. Make sure the slide sit on a flat surface for 15 seconds or so to let the cell particles settle down. This will make the focusing much easier.

-insert the slide ( the chamber with sample first in, cover slide up)

-Autofocus light on, press up or down arrow to have the cells in focus. Use magnify at 4X for better view.

Protocol parameters:

-use noise reduction level to adjust the sensitivity

-live cell sensitivity to adjust miss called cells

-size limit to exclude debris

-circularity: perfect round cell is 100, 1 for definitely not round