Cell cycle assays are applied in early toxicity testing and in screening for anti-cancer agents. One of the most important aspects in anti-cancer treatments is the inhibition of cell proliferation and cell division.

BCell cycle assays are applied in early toxicity testing and in screening for anti-cancer agents. One of the most important aspects in anti-cancer treatments is the inhibition of cell proliferation and cell division.

Both events can be analyzed using High Content Screening (HCS) approaches by multiplexing cell cycle-specific cellular events. Examples are BrdU or EdU staining, which detect the S-phase of the cell cycle by incorporating the nucleoside analog Uridine into newly synthesized DNA strands.

There are also well validated protein markers that are associated with certain cell cycle phases. One example is the phosphorylated histone H3 (pHH3), which is a common M-phase marker. DNA histogramming is another basic analysis tool which transfers very well to image based automated analysis of DNA content per cell.

High Content Screening can quantify all the cell cycle analysis readout parameters which are used in flow cytometry, the traditional tool for cell cycle analysis. Cell cycle analysis relies greatly on statistics over a large number of cells so to get reliable data it is essential to acquire multiple image fields per sample combining data from several thousand cells. A large number of cells must be analyzed in a short period of time in order to achieve an acceptable screen throughput.

The figure above shows a schematic representation of the cell cycle phases, incorporation of EdU and pHH3 detection. The cells have been stained with a nuclear marker (blue), an antibody for EdU (green) and an antibody for pHH3 (red) in various stages of the cell cycle.

Both events can be analyzed using High Content Screening (HCS) approaches by multiplexing cell cycle-specific cellular events. Examples are BrdU or EdU staining, which detect the S-phase of the cell cycle by incorporating the nucleoside analog Uridine into newly synthesized DNA strands.

There are also well validated protein markers that are associated with certain cell cycle phases. One example is the phosphorylated histone H3 (pHH3), which is a common M-phase marker. DNA histogramming is another basic analysis tool which transfers very well to image based automated analysis of DNA content per cell.

High Content Screening can quantify all the cell cycle analysis readout parameters which are used in flow cytometry, the traditional tool for cell cycle analysis. Cell cycle analysis relies greatly on statistics over a large number of cells so to get reliable data it is essential to acquire multiple image fields per sample combining data from several thousand cells. A large number of cells must be analyzed in a short period of time in order to achieve an acceptable screen throughput.