Making fluorescent intensity equivalent across images

[jeslea]

Hi, I have a batch of fluorescent images which I would like to perform cell counts on. I noticed that the background intensity differs across images. In order for the counts to be comparable across images I would like to adjust all the images so that the level of background staining is equivalent. Is there a way to do this? Is there a way to adjust the intensity of the entire image until the background reaches a specified value which I will keep constant across images?

Thank you!

[samjlord]

jeslea wrote

Hi, I have a batch of fluorescent images which I would like to perform cell counts on. I noticed that the background intensity differs across images. In order for the counts to be comparable across images I would like to adjust all the images so that the level of background staining is equivalent. Is there a way to do this? Is there a way to adjust the intensity of the entire image until the background reaches a specified value which I will keep constant across images?

Under Image -> Adjust -> Brightness/Contrast, click "Set" and select "Propagate to all open images." Then all images will have the same display scaling.

-Sam

[GuyM]

Hi
You may find the "subtract background" command useful for your needs:

http://imagejdocu.tudor.lu/doku.php?id=gui:process:subtract_background

I think that it re-calculates the background for each slice, but even if not, implementing this in a macro should be quite simple.

Guy

[jerie]

Hi Jeslea,

Image -> Adjust -> bleach correction 'simple ratio' might do the job.

Abraços,Jens

Dr. Jens Rietdorf, visiting scientist @ center for technological
development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro
Brasil.

On Thu, Aug 25, 2016 at 3:52 AM, jeslea <[hidden email]> wrote:

> Hi, I have a batch of fluorescent images which I would like to perform cell
> counts on. I noticed that the background intensity differs across images.
> In
> order for the counts to be comparable across images I would like to adjust
> all the images so that the level of background staining is equivalent. Is
> there a way to do this? Is there a way to adjust the intensity of the
> entire
> image until the background reaches a specified value which I will keep
> constant across images?
>
> Thank you!
>
>
>
> --
> View this message in context: http://imagej.1557.x6.nabble.
> com/Making-fluorescent-intensity-equivalent-across-images-tp5017113.html
> Sent from the ImageJ mailing list archive at Nabble.com.
>
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> ImageJ mailing list: http://imagej.nih.gov/ij/list.html
>

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