Re: Help needed with method of background substraction

Posted by Djoere Gaublomme on Nov 05, 2014; 3:16pm
URL: http://imagej.273.s1.nabble.com/Help-needed-with-method-of-background-substraction-tp5010244p5010318.html

I think you might use 'Process'-->'Math'--> 'Subtract' and enter the  
value you would otherwise use for your Min settings to remove the  
background.

I would just remark though, that when using signal intensity as an  
output/measurement, you really have to stop and think about your setup  
(is it possible to use the same lighting conditions for all samples  
when taking the pictures? , is it possible to always Subtract the same  
value ?, etc.), otherwise your numbers might be rubbish.

greets

Djoere


Quoting khanas <[hidden email]>:

> 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
>
>
>
> --
> View this message in context:  
> http://imagej.1557.x6.nabble.com/Help-needed-with-method-of-background-substraction-tp5010244.html
> Sent from the ImageJ mailing list archive at Nabble.com.
>
> --
> ImageJ mailing list: http://imagej.nih.gov/ij/list.html

--
ImageJ mailing list: http://imagej.nih.gov/ij/list.html