Phenotypic Screen for Mitotic Inhibitors

This causes the accumulation of round-shaped cells with intense staining of the DNA and microtubules, which can be detected by fluorescence staining using antibodies or dyes. The spindle-assembly checkpoint in cells that are arrested in mitosis is inactivated only after proper error correction. Alternatively, cells commit suicide by apoptosis. 

High-content screen (HCS) is a phenotypic screening that monitors multiple cellular parameters simultaneously. HCS employs fluorescence-based reagents (antibodies, dyes that bind or localize to a given cell component, sensors) to generate a multicolor fluorescence readout that is usually recorded using automated optical image acquisition devices. 

Triton X-100). Alternatively, cells can be fixed and permeablized with organic solvents such as methanol and acetone. The type of fixative depends on the cellular components that need to be detected and the employed antibodies. Cellular components are then detected either directly by means of dyes (e.g., fluorescent dyes that bind to and stain DNA) or antibodies that are labeled with fluorophores. Alternatively, an indirect detection can be performed by means of primary antibody that specifically binds to a protein of interest and a secondary antibody that recognizes the primary antibody and is labeled for detection (usually with a fluorophore). Finally, samples are analyzed by means of fluorescence microscopy. 

O Control wells O Normal phenotype t Mitotic phenotype 

The action of a given compound is dose dependent if the influence of the compound in the studied system (e.g., on cells) changes when the concentration of the compound is changed. 

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