Immunohistochemistry IHC Endothelial Staining - Pixel Count Semi Quantification Query

[AHTONG]

Dear NABBLE Community,

I am in dire need of some guidance in terms of how to semi quantify the endothelial lumens you can see in the attached image. I have tried to look for literature explaining how to semi quantify the pixel intensity of the whole brown IHC stained endothelium. However, I have only been able to quantify small areas within the endothelium consistently and extrapolate a pixel intensity.

I wanted to know if use Image J has the ability to measure the pixel intensity of the BROWN endothelial capillary stain between two freehand drawings around the outer and inner edges. Any other solutions to my question would be greatly appreciated. I ideally would like to measure the pixel intensity of the whole lumen and standardize that reading by dividing over the area measured.



Thank you,
Andy