The Comet Assay module is dedicated to the Single Cell Gel Electrophoresis (SCGE) analysis commonly referred to as the Comet Assay: it incorporates all the tools needed to characterize nuclei.

The experiment

Analyses of slides from a given experiment are grouped into a protocol which defines the name, the test compound, its concentration and the date of analysis. An experiment can be performed over several days and can be organized in the form of a directory (created automatically according to the experiment name) in which the analytical protocol, images and results are saved.

Acquisition and calibration

The user can choose from the whole set of image acquisition tools (color or B&W) and calibration tools (up to 10 objectives). The use of a motorized stage is possible, allowing analysis of the entire exploration zone.

Automated measurement

The software automatically detects the locations of the various comets. The user can then validate the different measurements in real time. He or she can choose the site for background above and/or below and at any moment can graphically observe the detected head and tail outlines on the image. The user can modify the measurement zone for the comet and the background, as well as the detection parameters.Two graphics (2D and 3D profiles) enable the user to better visualize the various intensities (comet, head, tail and background) and potential saturation.

Results

All the results are saved in an Excel compatible spreadsheet:
- Comet: surface, DNA quantification.
- Head: surface, radius, % DNA in the head, average gray scale and standard deviation. - Tail: length, % DNA in the tail, % tail DNA/total DNA.
- Derived parameters: Tail Moment, Olive Tail Moment, Tail Moment Arm, etc. The measurement set is then analyzed to generate a series of statistics (mean, number, total, standard deviation, SEM, minimum and maximum).