Komet Software for Comet Assay - Functionality

Rapid Scoring, Multi-session Capability & Automatic Data Organization

• Once a scoring session is in progress, speed is of the essence. The user acquires and scores cells quickly with a single right mouse click.
• Komet toolbar allows rapid and convenient program control.
• Scoring sessions can be suspended and resumed at a later time with the session manager.
• Background correction for every cell scored and automatic detection of head and tail extent. Instantaneous computation of all parameters.
• Immediate visual feedback of analysis results, on image, in data summary and in Analysis Graphics.
• Head DNA, Tail DNA, Tail Length, Olive and Extent Tail Moments and L/H (Tail Length to Head Diameter Ratio) are presented in the Key Results panel.
• 24 parameters are computed from the comet image based on intensity and migration patterns.
• Comet integrated intensity, moment, skew and inertia are included.
• User can over-ride to interactive scoring on-demand - especially useful for “hedgehog” or heavily damaged comets.
• Cell saturation check prevents unsuitable analysis and warns you to adjust the camera or illumination.

Figure 3 - "Key Results" are tabulated below the Image Window as shown above. In the Comet Options setup phase theyare colored black. These examples show typical key parameter values for a heavily damaged comet and slightly damaged comet respectively. Heterogeneous comet morphology is common within a single sample, so statistical methods are applied to data from approximately 100 cells (randomly selected) per specimen. As with any scientific experiment, good experimental design plays an important part in the reliability of conclusions.

Figure 4 - Scoring. A live image appears in the image Window (shown pseudo-colored here). A right mouse click initiates capture and analysis for DNA migration. Intact DNA remains in the comet ‘Head’, damaged DNA migrates to the ‘Tail’. Vertical calipers (in red) show the extent of the Head and Tail. Courtesy Marie Vasquez Helix3 Inc.

Results

Scoring From File

• Standard and proprietary 8, 12, 16 bit gray and 24 bit color files are handled seamlessly
• Komet automatically loads files from a user- defined directory on demand
• "Flip or Rotate on Open" function allows images (with comet tails to left) to be automatically oriented for analysis
• Apply "Calibration on Open" calibrates images for real measurements
• Automatic Pop-up Open button minimizes mouse movements and reduces operator fatigue
• Analysis and presentation of results proceeds as in Live scoring

Figure 5 - The Analysis window shows comet intensity profiles and 24 comet parameters are recorded in a data grid. The integrated intensity profiles of comet, background, head and tail and can be useful for identifying background debris and comet overlap.