Help needed with method of background substraction

Posted by khanas on Oct 30, 2014; 3:32pm
URL: http://imagej.273.s1.nabble.com/Help-needed-with-method-of-background-substraction-tp5010244.html

Hi all,

Have come across an image processing issue this morning and would really
appreciate your help! Forgive my ignorance please I am quite new to
microscopy/imageJ!

Essentially to quantify nanoparticle uptake into cells we take images with a
whole cell fluorescent stain and a reflectance channel - the nanoparticles
turn up in the reflectance channel and we use the staining to draw around a
cell and analyse it for mean intensity.

To remove background reflectance other people in my lab have first split the
channels of the image, and then using the max and min controls in B&C
changed the min value until any background has been removed. After this they
have selected the area in question (using the aforementioned stain), and
then Measured mean intensity.

Unfortunately for some reason, potentially due to an update, it now doesn't
measure between the values we set. it only ever gives mean intensity of the
unedited/adulterated image and I have verified this by measuring the same
area a few times with different Max and Min settings.

If I save the file as another format this can work but such a compression
would be a problem from a quantification point of view!

A colleague just showed me this forum and I will have a look through to see if there are any existing solutions but any and all help would be very much appreciated!

Many thanks
Abs

Any help would be very much appreciated!

Thanks