problems using ImageJ!!

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Gina
Gina's picture
problems using ImageJ!!

So I need to quantify the intensity of fluorescence of an integrin staining I did with Cy2 conjugated Ab to the specific mAB....I was told to use Image J...problem is I cannot figure out the program for the life of me. I imported my pictures but then, I'm trying to select only the fluorescent areas and I don't know how. Even if I figure that out, how do I actually quantify intensity?? Help!!!

mahi
mahi's picture
please go through this
Arvind Singh Pundir
Arvind Singh Pundir's picture
Gina wrote:So I need to

Gina wrote:

So I need to quantify the intensity of fluorescence of an integrin staining I did with Cy2 conjugated Ab to the specific mAB....I was told to use Image J...problem is I cannot figure out the program for the life of me. I imported my pictures but then, I'm trying to select only the fluorescent areas and I don't know how. Even if I figure that out, how do I actually quantify intensity?? Help!!!

Hi Gina
hope your problem is solved by now, but as this post is revived by one of the member i would suggest to all ImageJ users to subscribe to imageJ mailing list  for all your querries, discussion related to ImageJ
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heehawmcduff
heehawmcduff's picture
I'm not sure if this will

I'm not sure if this will help you but some of the principles may work with your application.  I used this method for densitometry.
http://stanxterm.aecom.yu.edu/wiki/index.php?page=Using_ImageJ
and this may help for fluorescence
http://knol.google.com/k/liang-cai/how-to-use-imagej-to-quantify/3ez005w9gduud/4

The FFM
The FFM's picture
Sorry I did not see this

Sorry I did not see this sooner.
Open your image in Image J and use the selection tools to draw boxes or another shape of outline around the region of interest.
In the "Analyze" menu select "Set Measurments"
In the window that opens you can select all the parameters youy want to measure in the selection area.
So for example you would select Mean Gray value, Min & Max Gray Value, Area and Integrated density.
Click OK and now go back to your image with the selected area.
 
Click Ctrl M (Apple M on a Mac) and it will measure all those values you selected above and paste them into a results table that you can export to excel.