Cell Profiler software: An easy screening tool for DNA damage estimation in fish erythrocytes from comet assay image
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DNA fragmentation by single strand breaks (SSBs) or double strand breaks (DSBs) is major concern in genotoxicity research. DNA damages can be easily known through comet assay or single cell gel electrophoresis (SCGE). Since decades, scoring through software for DNA damages in images have been developed by researchers. These softwares depend upon manual scoring on individual comet in a particular interface. The evaluation under software may have biasness and error during scoring by each researcher and few softwares are unable to access easily because many of these are commercial products. However, CellProfiler (CP) image analysis software (Version 2.1.0) is free, easy operation, faster and automated screening by computer itself. An attempt was made to detect DNA damages mainly comet scoring through CP software as whole comet, comet head and comet tail from image of previously studied single cell gel electrophoresis (SCGE) in the peripheral erythrocytes of fish exposed to benzene as experimental image. The results particularly on length and area of whole comet, head and tail were obtained after automated analysis in the CP software. The image processing study was done of the objects present in fluorescence microscopy image to know maximum DNA damage at each cell level for the fish erythrocytes. It was concluded that the present study of image based screening for DNA damages as details of comet and its head and tail evaluation by shape and area in the fish erythrocytes can be a suitable tool for genotoxicity prediction along with risk assessment at DNA level. The shape descriptor as Zernike moments order 0 to 9 can also be suitable parameters to know accuracy of the shape of comet and its head and tail in the image. Finally, high-throughput automated screening of comet test can help in disease diagnosis and repair mechanisms as well as environmental monitoring of genotoxin(s) within short period of time.
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