Immunohistochemistry Quantification

[pipette_man]

Hello guys,

I am having problems quantifying of my immunohistochemistry results.

I am getting significance when I see the results, however with Image J it shows no difference.


Is this due to the fact that Image J is not sensitive enough?

What do you guys think?

thanks in advance

[inmuno]

Hello:
What are you quantifying? cells,areas,nucleus... fluorescence or chromogen? I have used ImageJ ocasionally, normally we use the AnalySIS program but they are similar. The problem could be the tresholds, the ROI selecction or for example the size of the cells if you are quantifying areas (not the same in all the samples/zones), the quality of the images...
I don´t know if i can help you...but i need more information to try...
My opinion is that if you see evident diferences between samples under the microscope ,the program should detect them if all the settings and the images are ok.
Best regards

[radam2k]

I assume that you are attempting to quantify the expression of a nuclear, membranous, or cytoplasmic marker. Please provide more information, including which marker you are studying, which area you are evaluating (e.g., an area of interest versus the whole slide), and what quantitative threshhold you are employing.

[suzan]

I have the same problem, as I need some help on how to use image J to quantify my immunohistochemistry results as I need to quantify the intensity of the florescence staining in my slides?!!