Comet assay shows a fragmentation of DNA. The measure must be done in a TAIL .The size of the comet tail reflects oxidative DNA damage by the extent of the rupture of the DNA helices and can be quantified using image intensification and computational analysis methods. (Ladeira MSP, Rodríguez MAM, Salvadori DMF, Queiroz DMM, Freire-Maia DV (2004) DNA Damage in Patients Infected by Helicobacter pylori. Cancer Epidemiol Biomarkers Prev 13:631-7.). I hope it help you.
Thank you very much for your answer. indeed all your answers are right regarding to comet assay measurements but my question was is comet assay software IV( http://www.perceptive.co.uk/cometassay/) can be used to measure DNA% in tail and head?
thanks again,
Hi Sarah, I am using other SW, nevertheless all comet SW are about drawing histogram from comet pixel intensities to delineate head and tail. So I expect Perspective Comet Assay IV provides these readings as a standard output. If you are shopping around, far better SW with semi-auto, and when you have microscope equipped with motorized table also automatic reading is LUCIA Comet Assay of www.lim.cz. Best, karel
Karel,
Thank you a million for you respond. it was really helpful. I looking for good software to buy and finish my project and I feel confuse with many software around. have you tried the software you talked about in your respond? Does this software is expensive?
Hi Sarah, I am using LUCIA long way (nearly 20 years) back when I found Kinetics Imaging, that time prime choice, awkward and hostile to use. LIM company is primarily teamed with Nikon and officially cover all their image grabbing needs. I am not aware about current pricing of any comet SW, but you should contact companies. Usually they give trial periods to make up decision what is the right cup of tea before purchase. Many 'cometeers` are using free NIH SW. I have never looked for it (no need), but I am aware about this option. Good hunt!-)))
Hi Sarah, I am using Comet IV from Perspective. The output includes % tail intensity, tail length and tail moment. It comes also with a spreadsheet macro with several options to Choose from. For example can you choose toe get a histogram and box plot overview of your comet scores.
However, I have no idea about the pricing. Best Andrea
I believe Perceptive Comet IV is the most widely used system. It will cost around 10.500 Euro with camera interface card. Most other programmes will do the same so if money is an issue you may look elsewhere. We recently purchased Andor Komet 6 for 3400 Euro w/o camera. We got a small discount of 400. You may also try visual class scoring, which is free of charge, and Works very well. It has been documentet to give similar results as image analysis.
The Alkaline Comet Assay: Towards Validation in Biomonitoring of DNA Damaging Exposures. By Peter Møller in Basic & Clinical Pharmacology & Toxicology 2006, 98, 336–345.
Genotoxicity of Environmental Agents Assessed by the Comet Assay. By Peter Møller in Basic & Clinical Pharmacology & Toxicology 2005, Vol 96 supplement I p1-42.
I can send them to you if you send your e-mail adress to nrj at nrcwe.dk.
Best regards Nicklas
Thank you so much for responding. is komet 6 good software for picture analysis?I have may picture ready but we do not have software my adviser try to buy best software for pictures. We have two camera which just take pictures.I will send you an email from my university account.
Thank you again
Komet 6 works fine, and it is cheaper than Perceptive. The cheapest option is still the No-Software image analysis as used in the papers I mentioned. Basically, they have a printed example of nuclei with no damage (category 0), Very low damage (category 1), etc etc to highest possible damage (category 4). They have tested how different people assign comets on the screen (or images) to the categories and compared this with software scores. Basically there is no difference in the results only in what statistics you can use. Ones you get well used to the system, you do not even need the reference Pictures. You know instantly what category is right.
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