Automated comet assay analysis

✍ Scribed by W. Böcker; W. Rolf; T. Bauch; W.-U. Müller; C. Streffer

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 367 KB
• Volume: 35
• Category: Article
• ISSN: 0196-4763
• DOI: 10.1002/(sici)1097-0320(19990201)35:2<134::aid-cyto5>3.0.co;2-9

✦ Synopsis

Background: Recently the ''comet assay'' or ''single-cell gel electrophoresis assay'' has been established as a sensitive method for the detection of DNA damage and repair. Most of the software now available to quantify various parameters for DNA damage requires the interaction of a human observer. In this report, we describe an automated analysis system that is based on self-developed software and hardware and needs minimal human interaction. Methods: The image analysis is divided into two parts: 1) automatic cell recognition and comet classification and 2) quantification of desired comet parameters. Image preprocessing, segmentation, and feature classification were developed with algorithms based on mathematical morphology. To enhance evaluation speed, we have introduced parallel processing of data under the Windows NT operating system (Microsoft Corporation, Redmond, WA). Use of an analogue real-time autofocus unit (Bo ¨cker et al.: