Many papers that do spinal IHC in the dorsal horn superimpose boundaries on the laminae.
How do people consistently and accurately do this? What are the features or landmarks that you use?
I know that certain molecular markers define laminar areas, for example PKC-gamma and lamina II inner and IB4 as lamina II outer. But in the absence of these markers, how do people do it? Neuronal density? that means you need to always use NeuN or Nissl. But that then takes up a color channel.
What do you do? And what methods are there in the literature?