# How do people accurately and methodically draw laminar boundaries in spinal cord IHC images? **URL:** \ **Category:** Microscopy (IHC/ISH/ICC/In Vivo)\ **Created:** [October 31, 2017, 4:52pm UTC](https://forum.painresearcher.net/t/how-do-people-accurately-and-methodically-draw-laminar-boundaries-in-spinal-cord-ihc-images/318 "2017-10-31T16:52:19Z")\ **Posts on this page:** 1\ **Showing post:** 5
**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:** [October 31, 2017, 7:06pm UTC](https://forum.painresearcher.net/t/how-do-people-accurately-and-methodically-draw-laminar-boundaries-in-spinal-cord-ihc-images/318/5 "2017-10-31T19:06:57Z")
That’s extremely helpful Thanks Liz! I use fluorescent Nissl stain, but not all the time: > [@Stuff I tried during grad school/post-doc that won't get published but could be useful](http://forum.painresearcher.net/t/stuff-i-tried-during-grad-school-post-doc-that-wont-get-published-but-could-be-useful/302/14): > > For pan-neuronal labeling in the DRG, I’ve moved to Neurotrace, which is a fluorescent Nissl stain. The different colors have variable staining qualities. Here is Neurotrace Blue. I used 1:200 diluted in PBS for 10 mins at room temp. I do a quick wash in PBS and then mount. I’ve found the Far red Neurotrace to be good too. The Green Neutrotrace is very strong, but it also stains non-neuronal nuclei too (like real Nissl). I’ve found the blue and Far red to do th… I’ll start doing it again. Thanks for the annotation. --- _[View the full topic](https://forum.painresearcher.net/t/how-do-people-accurately-and-methodically-draw-laminar-boundaries-in-spinal-cord-ihc-images/318)._