Image J

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cooper's picture
Image J

Is anyone familiar with Image J from the nih website?

I would like to know the best way to analyze cells on immunocytochemistry-stained slides (using ABC method). This program seems to be able to give intensity of staining somehow so that I could interpret my staining in more that just a qualifying manner.

If anyone has any ideas, let me know. Thanks!!

samm's picture
Image J (or even programs

Image J (or even programs such as PhotoShop or jASC PSPro) can give total intensity values for a slide. However, the only way you can get quantitative data out of your slides is to have IDENTICAL tissue preps - consecutive sections thin enough to have the same staining pattern. Then you can use one as a negative control to subtract the background. However, this is very cumbersome, and it will be easy to goof (time of development etc). See if you can use an alternative technique - including ELISpot, In-cell Westerns (LiCor, using the IR dyes instead of peroxidase) etc - which are designed to give more quantitative data.

Fraser Moss
Fraser Moss's picture
You can draw your region of

You can draw your region of interest around the area of the image to want to quantify and then hit Ctrl + M to measure.

In the "Analyze" menu you can "Set measurements" to set what is output in the results window.

e.g. mean intensity, Min and max values, area etc.

You can also type Ctrl H on any region of interest to get the Histogram of the pixel intensities of the region of interest or the whole image.

this ImageJ web site has many tutorials or examples