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8
Introduction
Basic principle
of counting
ADAM-CellT is an instrument which counts mammalian cell DNA by staining
with a fluorescent dye, Propidium Iodide (PI) or Acridine Orange (AO).
PI does not enter cells with intact membranes or active metabolism.
In contrast, cells with damaged membranes or cells with inactive metabolism
are unable to prevent PI entering the cell. As a result, the nuclei of cell
membrane-damaged normal cells or non-viable cells will be stained.
Solution T
PI
is composed of the PI and cell membrane lysis buffer.
Since lysis buffer in Solution T
PI
changes intact cell membrane to damaged cell
membrane condition, both viable cells and non-viable cells can be stained. AO
included in Solution T
AO
is permeable dye which permeates cell membrane
and stains DNA. Regardless of the condition of the cell membrane or active
metabolism, AO can stain both viable cells and non- viable cells.
The ADAM-CellT provides two kinds of staining solutions: AccuStain Solution
T for the total cell counting and AccuStain Solution N for the non-viable cell
counting. AccuStain Solution T is categorized into Solution T
PI
and Solution T
AO
.
AccuStain Solution N for the non-viable cell counting is composed of the PI
alone. After staining samples, the prepared cells will be loaded into the chip.
The viability will be automatically calculated in the ADAM-CellT software after
each measurement of the total cells and the non-viable cells.
T Channel
A: Total cell / B: Non-viable cell
N Channel
AccuChip 4 Channel
AccuChip 2 Channel
*Viability(%)=
X 100
A
(A-B)
T1
N1
T2
N2
.