How to use photoshop to measure the amount of shades of Brown in a given area (Immunohistochemistry)

I try to use photoshop measure intensity area of brownish (some variance) in a part of the image of a mouse knee joint.  I have stained a particular marker in these samples and hoped I could quantify because she is extracellular and discreet

Thank you!

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Scott, I believe that I came up with a way for you to make selections within your settings and have prepared an example to demonstrate the method. Much depends on whether this technique, using your spot color and a specimen, will work. If so, you can measure the number of pixels in the selection following the instructions I provided previously. Here goes:

1. This sample represents the darker brown spot.

2 build a scale of gray similar to this (I can provide one shown here)

3 Edition > fill with your Brown sample more dark and set its blend Mode to color. (When I used the sample in step 1, this was the result.)

4 attach it to your specimen as shown image file here. (I have not colorized that half of the test image to demonstrate the next step only select the sample Brown even if the black & white area has the same tone value.)

5. with the magic wand, a tolerance of zero and contiguous unchecked value, when you click on a box in the balance its corresponding value will be selected in the image. Here is an example. The histogram can then be used for a number of pixels from all selected areas. Repeat for each sample in your settings.

I hope it does the job for you.

[(Edit: quand vous arrivez àles valeurs plus sombres, vous pouvez constater qu'augmenter la tolérance contribue à la couverture de l'écart entre valeurs.)]

Tags: Photoshop

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