How to quantify fluorescence intensity with image J. (please HELP)

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Polikarp
Polikarp's picture
How to quantify fluorescence intensity with image J. (please HELP)

 
Hi there,
I was wondering if anyone can tell me how to quantify fluorescence intensity with image J. I don't need to analyze single cells, I just want to measure the fluorescent intensity of an image (or selected area of an image) and compare with others taken under same conditions.
According to the protocol I know that I have to do the following:

  1.  

    Convert the color picture to 8it-b black and white scale.

  2. Subtract the background fluorescence equivalently for all images by setting the threshold to 50% maximum intensity.
  3. To quantify the mean fluorescence

I will appreciate if someone can post a tutorial with screen-shots regarding my question.
Attached is a picture fro analysis.
Thank you in advance for your help

christiandersen
christiandersen's picture
If you are having a hard time

If you are having a hard time using Image J may i propose that you have a look at the www.imagemeasurement.com and download the free version og KLONK Image Measurment Light, which has been made for intuitve usage for exactly what you descibe.

waklim
waklim's picture
Hi there, I know this post is

Hi there, I know this post is quite a while Polikarp, but I wonder how is the process you did in measuring the intensity?Can you share with me? Thank you.