4
Introduction
Basic principle
of counting
ADAM-MC2 is based on staining mammalian cell DNA with a fluorescent
dye, Propidium Iodide (PI). PI does not enter cells with intact membranes
or active metabolism. In contrast, cells with damaged membranes or with
inactive metabolism are unable to prevent PI entering the cell. As a result,
the nuclei of non-viable cells will only be stained. The ADAM-MC2 pro-
vides two kinds of staining solutions. AccuStain Solution T for the total cell
counting is composed of the fluorescent dye (PI) and lysis solution. Ac-
cuStain Solution N for the non-viable cell counting is composed of the
fluorescent dye and PBS.
In order to measure the total concentration of cells, the plasma mem-
branes of all the cells must be disrupted to stain all the Nuclei with PI. The
process of disrupting and staining is achieved by treatment with AccuStain
Solution T.
In the second solution, live cells remain intact and are not stained.
Only the non-viable cells are stained and detected. After treatment, the
prepared cells will be loaded into the chip. The viability will be automati-
cally calculated in the ADAM-MC2 software after each measurement of the
total cells and the non-viable cells.
Add PI
T Channel
A: Total cell / B: Non-viable cell
N Channel
AccuChip 4 Channel
AccuChip 2 Channel
*Viability(%)=
X 100
A
(A-B)
Total cell count
Non-viable cell count
Non-viable cell
Non-viable cell
Viable cell
Viable cell
PI-stained nuclei
PI-stained nuclei
PI-stained nuclei
T1
N1
T2
N2
Add PI + Iysis sol.