# Quantification of skin innervations **URL:** \ **Category:** Uncategorized\ **Created:** [February 26, 2018, 3:02pm UTC](https://forum.painresearcher.net/t/quantification-of-skin-innervations/405 "2018-02-26T15:02:14Z")\ **Posts on this page:** 7\ **Page:** 1
**Author:** ![Ewa-RUD](https://avatars.discourse-cdn.com/v4/letter/e/85f322/32.png) [@Ewa-RUD](https://forum.painresearcher.net/u/Ewa-RUD)\ **Post date:** [February 26, 2018, 3:02pm UTC](https://forum.painresearcher.net/t/quantification-of-skin-innervations/405/1 "2018-02-26T15:02:14Z")
Hi Guys, How would you quantify skin innervation from images? My idea was to carry out thresholding image analysis. Would anyone have a better idea? ---
**Author:** ![achamess](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/achamess/32/534_2.png) [@achamess](https://forum.painresearcher.net/u/achamess)\ **Post date:** [February 26, 2018, 3:10pm UTC](https://forum.painresearcher.net/t/quantification-of-skin-innervations/405/2 "2018-02-26T15:10:58Z")
What are you trying to quantify specifically? ---
**Author:** ![Ewa-RUD](https://avatars.discourse-cdn.com/v4/letter/e/85f322/32.png) [@Ewa-RUD](https://forum.painresearcher.net/u/Ewa-RUD)\ **Post date:** [February 26, 2018, 3:16pm UTC](https://forum.painresearcher.net/t/quantification-of-skin-innervations/405/3 "2018-02-26T15:16:19Z")
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. ---
**Author:** ![achamess](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/achamess/32/534_2.png) [@achamess](https://forum.painresearcher.net/u/achamess)\ **Post date:** [February 26, 2018, 3:21pm UTC](https://forum.painresearcher.net/t/quantification-of-skin-innervations/405/4 "2018-02-26T15:21:09Z")
@tberta? @liz What do you think? ---
**Author:** ![tberta](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/tberta/32/145_2.png) [@tberta](https://forum.painresearcher.net/u/tberta)\ **Post date:** [February 26, 2018, 4:29pm UTC](https://forum.painresearcher.net/t/quantification-of-skin-innervations/405/5 "2018-02-26T16:29:36Z")
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. ---
**Author:** ![Ewa-RUD](https://avatars.discourse-cdn.com/v4/letter/e/85f322/32.png) [@Ewa-RUD](https://forum.painresearcher.net/u/Ewa-RUD)\ **Post date:** [February 27, 2018, 12:29pm UTC](https://forum.painresearcher.net/t/quantification-of-skin-innervations/405/6 "2018-02-27T12:29:56Z")
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 🙂 ---
**Author:** ![liz](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/liz/32/267_2.png) [@liz](https://forum.painresearcher.net/u/liz)\ **Post date:** [February 28, 2018, 3:40pm UTC](https://forum.painresearcher.net/t/quantification-of-skin-innervations/405/7 "2018-02-28T15:40:05Z")
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: > > > Vincristine (VNC) is commonly used to treat pediatric cancers, including the most prevalent childhood malignancy, acute lymphoblastic leukemia. Although clinical evidence suggests that VNC causes peripheral neuropathy in children, the degree to which pediatric chemotherapeutic regimens influence pain sensitivity throughout life remains unclear, in part because of the lack of an established animal model of chemotherapy-induced neuropathic pain during early life. Therefore, this study investigated the effects of VNC exposure between postnatal days (P) 11 and 21 on mechanical and thermal pain sensitivity in the developing rat. Low doses of VNC (15 or 30 μg/kg) failed to alter nociceptive withdrawal reflexes at any age examined compared with vehicle-injected littermate controls. Meanwhile, high dose VNC (60 μg/kg) evoked mechanical hypersensitivity in both sexes beginning at P26 that persisted until adulthood and included both static and dynamic mechanical allodynia. Hind paw withdrawal latencies to noxious heat and cold were unaffected by high doses of VNC, suggesting a selective effect of neonatal VNC on mechanical pain sensitivity. Gross and fine motor function appeared normal after VNC treatment, although a small decrease in weight gain was observed. The VNC regimen also produced a significant decrease in intraepidermal nerve fiber density in the hind paw skin by P33. Overall, the present results demonstrate that high-dose administration of VNC during the early postnatal period selectively evokes a mechanical hypersensitivity that is slow to emerge during adolescence, providing further evidence that aberrant sensory input during early life can have prolonged consequences for pain processing.