DNA Damage Applications (Micronucleus Assay)
Introduction
The iGeneration systems, with automation capabilities and enhanced software, may be used to detect and quantify micronuclear formation. Evaluation of clastogenic effects of drugs is usually accomplished by some form of the micronucleus assay, where broken or detached chromosomes are separated from the spindle apparatus. After cells undergo mitosis, the fragments become trapped in the cell cytoplasm and form micronuclei.
Highlights of Analysis with iGeneration Imaging Cytometers
The iNovator Software Development Toolkit allows you to automate a cell-line micronucleus assay. The following figure illustrates the basic strategy used to identify cells, nuclei and micronuclei in order to quantify micronuclear formation. The assay is done in multi-well plates examining a titration series of a clastogenic drug such as etoposide.

In panel A, cells are identified using a whole-cell marker. The watershed algorithm identifies the nuclei of all cells, including those that are binucleated, as well as any associated micronuclei. Binucleated cells result in actively dividing cells but do not advance, as cytokinesis is blocked by cytochalasin. This allows for identification of cells that are actively growing and dividing. Using a strategy where specific populations are identified, binucleated cells are identified and associated micronuclei are quantified. An example of overall data analysis is seen below.
Benefits of Using iGeneration Technology
The optional iNovator Application Development Toolkit makes it possible to identify closely spaced objects such as nuclei in a binucleated cell, as well as the associated micronuclei. Multiple probes can be analyzed in a single sample while obtaining widefield images, which may be correlated to the quantitative data in galleries or field images. The iGeneration platform combines the advantages of flow cytometry and image analysis in a single system. The iGeneration inverted format supports a diversity of carrier types, allowing flexibility in matching the needs of your application with the appropriate carrier type.
More Information

For more information, review the CompuCyte Published Materials and CompuCyte Bibliography.
Request the Micronucleus Assay Application Note.

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