ADAM-MC™ Automatic Cell Counter

Technology

The ADAM MC automated cell counter performs viability and cell counting measurements using the PI staining method of dead-cell staining combined with advanced image analysis. ADAM, is the new standard of automatic cell counting, based on a fluorescent microscopy technique for counting cells. The ADAM utilizes sensitive fluorescence dye staining, LED optics and CCD detection technologies to make the cell analysis more accurate and reliable. To count cells using ADAM, the cells are mixed with a Propidium Iodide (PI) stain and directly pipetted on to a disposable plastic chip. The chip is then loaded onto a precision stage. An ADAM system is automatically focused onto the chip and cells that have been stained are recorded by a sensitive CCD camera. The image results are automatically processed generating the cell count which is displayed on the front of the instrument. Simple. Fast. Accurate. Reliable.

Procedure

As accurate as FACS counting Cell counting accuracy depends on the counting volume. ADAM analyzes 60 images which is equivalent to an 8.5 ul counting volume. 8.5 ul counting is 8.5 times more accurate than a 1 ul analysis. These improvements specifically address variation attributable to low sample reads (some competitor models only perform one read ), low sampling volumes ( most models only read 1ul or less ), and inaccurate resolution of cells (often found when cells clump together ). The results are amazing and it is common to have R2 values over 0.99 and CVs under 1%.

New technology behind the counting accuracy of ADAM-MC

After the samples are stained with a fluorescent dye, propium iodide, which intercalates DNA to stain the nucleus of target cells, fluorescent images are taken automatically. The system takes the 22-60 images by the movement of an automatic X-Y stage. The obtained images are processed by image analysis software integrated inside the system. All procedures are fully automatic once the stained samples have been dropped to the disposable chip and the chip is loaded.

• LED excitation
• Precision automatic stage
• Automatic focusing
• Sensitive CCD detection
• Automatic image analysis
• Disposable microchips
• Large counting volume guarantees accurate results

Automatic cell counting device, ADAM!

Propidium Iodide Staining

ADAM-MC is based on staining mammalian cell DNA with a fluorescent dye, Propidium Iodide (PI). PI does not enter cells with intact cell 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. ADAM-MC provides two kinds of staining solutions. AccuStain Solution T for the total cell counting is composed of the fluorescent dye (PI) and lysis solution. AccuStain 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 membranes 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 automatically calculated in the ADAM-MC software after each measurement of the total cells and the non-viable cells.

Accuracy and Repeatability

Samples with low, medium and high concentrations of cells counted with ADAM-MC.	Correlation of total cell counting between hemocytometer and ADAM-MC using CHO cells.

Comparison of cell viabilities between ADAM-MC and flow cytometry. SK-OV, HeLa and NCl-H23 cells were treated with 100, 300M H2O2 for 3 hours, then analyzed by ADAM and flow cytometry. Values are given as means +-SD of three experiments.