Measuring the total fluorescence images

Hi everyone, I’m trying to measure the total fluorescence intensity on images of two animal models for comparison, I’m getting the area, mean of the gray values and the integrated density. I’m also getting measurements for background sections of images. can you help me? How should I use the data to make a graph of differences?. suggestions formulas. Thank you

The first step is to segment your image. Did you follow the “flexible workflow” for segmentation as explained here? http://imagej.net/Segmentation

I did a quick try with the data you posted. First converted to 8-bit, then thresholded using the default auto-threshold, and ended up with this:

This is a great illustration of Why (lossy) JPEGs should not be used in imaging!

I then applied a Median filter with radius 2. After that, the threshold using Huang looks like this:

From there, you can Create Selection, reset the threshold, then Revert the image to return to original RGB values:

And then Analyze :arrow_forward: Measure with the desired statistics set via the Analyze :arrow_forward: Set Measurements… dialog.

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