I did comet assay analysis using software open comet V1.3. I had no difficulty running the ImageJ program and got the result like the first picture attached. The problem started when I tried to open the output spreadsheet using Microsoft Excel 2010, when I first opened it, it showed message "the file you are trying to open is in a different format than specified by the file extension. Verify that the file is not corrupted and is from a trusted source before opening the file." (2nd picture) If I keep proceed with yes, It would be opened but for the first time I need to double click all the hashtag symbol in order to make the number appear (3rd picture). The number appeared for Comet intensity, head DNA, head DNA percent, tail intensity, tail DNA percent, tail moment, and olive moment were confusingly big like more than trillions (4th and 5th pictures). I don't think the number supposed to be this big. (or it is??)
Well my questions are How to get rid this problem? Is there anything I did wrong? I analyze the comet picture in JPEG format.
Thank you for helping since this the first time we do comet assay in our center.
Regards,
Raesa Yolanda
Dear Raesa,
yes, you're right, it is very big results. I can suggest you trying use another software, CASP. You can download it this link http://casplab.com/download. This software is more simple to use and it allows to get more correct results. If you will have problems with using it, you can ask me :)
Sincerely,
Maxim
Hi Raesa,
Is the output file a .CSV file? When the data file is opening, did it ask whether the file is comma-separated/tab-separated? If so, you can try the different options, which can affect the appearance and organization of the data in the corresponding excel cells.
On a different note, I have had good experiences using CometScore program.
Best of luck - let us know what works for you.
Best,
Jonathan
I did all my comet assay analysis using CometScore and Excel. I would recommend using this software.
Dear Maxim and Anne, thank you very much for the info, currently I'm interested in open comet because It analyzed the comet automatically, we don't need to choose the comet one by one like comet score and CASP, It save so much time and would reduce the bias.
Dear Jonathan, I have found the problem, You are right, it's about the decimal and thousand separator incompatibility between the software and excel. So I change the excel setting: office button> excel option > advance > decimal separator and thousand seperator (I change the decimal separator to (.) and thousand separator to (,) and problem solved.
Thank you all for the help.
For everyone who wish to use open comet, the download link and tutorial is available here:
http://www.opencomet.org/
Scientific publication:
OpenComet: An automated tool for comet assay image analysis; Redox Biology
Volume 2, 2014, Pages 457–465
http://www.sciencedirect.com/science/article/pii/S2213231714000032
Regards,
Raesa
Kindly suggest how to download a trial version of Comet score as I need this for trial.
Hi Raesa,
Sorry for the slow response, I just saw this post now. I'm happy that changing the decimal in Excel helped solve the display problem. To make sure that Excel shows the correct values, you can also directly look at the numbers calculated by OpenComet if you open the spreadsheet file in a text editor. Let me know if you have any other questions about OpenComet!
Best,
Ben
Hi everyone,
I am facing the same problem than Yolanda. So I tried to change the decimal and thousand separators and I could get rid of some presumably aberrantly high values (for Head and tail length f. ex.). The values for the Tail DNA %, tail moment and olive moment stay high though. And by checking the results in the text editor program I see that indeed the values seems to be super high. So is this correct than? And how would I proceed with percentages in the billion range?
Thank you for your help,
Nadine
Hi,
if you want to easily edit CSV files (such as those created by OpenComet) and to avoid the problems you mentioned I suggest using LibreOffice Calc, from the LibreOffice suite.
It is free and open source software and it allows you to open CSV files directly, you don't even need to tweak the import settings.
Regards,
Francesco
Benjamin M Gyori Could you please help me how to interpret the excel results in my thesis....I have used open comet software...single gel per sample...I am confused for whether to take the mean given in the excel output or the median..sometimes for single image I have per sample a single comet is detected with zero SD..nd mean....or i need to apply some sort of statistical test...Kindly guide me
Dear all,
we just had the same problem but could not solve it with changing the decimal and thousand separator. What we now did is using an older version of ImageJ, it was ImageJ 1.53t. We used OpenComet to analyze the images and opened the spreadsheet with LibreOffice. Then we saved the spreadsheet as an ".xlsx" file and opened it in Excel. This worked fine.
I hope this may help others. However, it would be good to correct this bug on the software side.
Best regards,
Vivian
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