We have HE stained samples from different animals and have noticed with the naked eye that there may be a difference in tissue density. Using ImageJ/Fiji, is it possible to measure the density of the tissue using an image?
My guess is we could gain data by measuring the strength of the stain, but would that necessarily tell us anything about the tissues density itself?
Thank you!
Dear Stephanie
Tissue density, as you put it, is a too broad term (it can mean nothing and everything). Clearing things a bit might help you get a good answer, for example:
What is the tissue you are trying to analyze?
Density of what exactly are you interested in (how many cells per field, density of collagen fibers...)?
Are all the slides from the same batch and stained by the exactly same protocol (staining can vary greatly depending on how were your slides processed)?
Nikola
Thanks for your responses! I'm analyzing scleral tissue from two different species, so my interest would be the density of collagen fibers. By naked eye, I've noticed that ones staining is more translucent than the other. The other is far darker, and simply looks denser. I would like to somehow quantitate this, but I'm not entirely sure how to approach this.
If by tissue density you mean that the staining is light for one and darker for other then you can select a rectangle region in each image and get a mean RGB value or even a histogram in grayscale or one of the R,G or B planes. The tissue with more gaps will have more white space and will have a mean value closer to 255 whereas the denser one will be darker due to less white space.
Note: Depending on your stain, you may even play with a different colorspace like HSV.
please i want to percentage or to quantify PCNA positive cells in seminiferous tubule image using imageJ
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