Hi,
Wonder if anyone can tell me how they quantify fluorescence intensity captured as .tif or .jpg files; i.e. is there any free software or shareware?
Basically, the worms with and without treatment showed difference in Nile Red fluorescence but I could not quantify the differences. Appreciate your input, thank!
p/s: I have tried different method to quantify, such as lipid extraction but the variation of results are a lot greater whereas the fluorescence intensity is more consistent.
Hi,
you could use ImageJ (free download http://rsbweb.nih.gov/ij/). I have already used this software to quantify fluorescence in cultured mammalian cells. tiff or jpeg formats should both be compatible with it.
Another software is Metamorph, but I think it’s not for free.
Good luck!
If you use the AxioVision software from Zeiss, you can quantify fluorescence as well.
Thanks, y`all for the resources you kindly suggested!
Rosina, I have tried out with Image J. However, theres no specific application for fluorescence measurement. Did you use the "Integrated Density" value as fluorecence intensity? If you dont mind, hope you could email me on exactly how you did it. Thanks a lot!
You can search the mailing list archives, and post specific questions there for more answers.
http://rsb.info.nih.gov/ij/list.html
Hi,
I hope it’s not too late.
When I used ImageJ I used to take the “Mean Gray value” of an area as an indicator of fluorescence intensity. In “Set Measurements” you can check “Area”, “Min & Max Gray Value” and “Mean Gray Value”. Then, you select an area in your image (by drawing it) and in “Analyze”, “Measure” you will find your data. I used to take these values to compare the fluorescence intensity between marked cells.
I hope this would help.
Best wishes!