I stained my skin samples for PGP9.5. I need to compare the innervation of WT and KO skin samples. I see that there is a difference but I need to quantify what how much fibres I see in the KO compare to the WT samples.
We incubate our skin sections with the pan neuronal marker PGP9.5 (Rabbit, 1:1000; AbD Serotec, Oxford, United Kingdom) along with anti-Collagen IV antibody (Goat, 1:100; Southern Biotech, Birmingham, AL). we determine the intraepidermal nerve fiber density by measuring the total number of fibers that crossed the collagen-stained dermal/epidermal junction into the epidermis (IENFs/mm). DAPI should also be able to indicate the different stratification of the skin. Fiber length is also often used to determine the intraepidermal nerve fiber density.
I saw a few papers that people talk about crossing of the fibres to the epidermis but I don’t necessarily know what that means as I can’t see the “crossing” itself. I see thicker fibres in the dermis and then thin one in the epidermis but I don’t understand how I am supposed to count the total number of fibres that crossed to the epidermis. Thanks a lot for your input
A graduate student in our lab recently did this kind of quantification, specifically the “crossing over” of fibres to the epidermis. She stained sections with both PGP9.5 and Collagen IV to delineate the epidermal-dermal junction, as Temo said. More details can be found in her publication below:
