Hello,
I stained mouse spinal cord sections using Nissl stain (cresyl violet). I am wondering how I can count for neurons, and differentiate between neurons and glia. I have attached an image taken using a microscope.
As an additional question: is the attached picture a good Nissl stain?
Thank you,
Suzie
I can´t see your image, but i used nissl stain in cerebral cortex of rabbits, to evaluted neuronal damage, there are many morphological differences between glial cells and neurons
https://www.sciencedirect.com/science/article/pii/B9780323449410000023
https://histologyguide.com//slideview/UCSF-163-spinal-cord/06-slide-1.html?x=19474&y=11223&z=25.8
look their quantitive observations with IHC: https://www.sciencedirect.com/science/article/abs/pii/S0147957119301845
Is there a reason why you use the Nissl stain instead of specific markers for neurons and glia (e.g. NeuN and GFAP)? This would probably facilitate automated analysis. In general, a fast way to count cells in large histological images is using a maxima finder, though I'm not sure if this would work well on Nissl or not. For questions on image analysis I can recomment you the image.sc forum. Here is for example a discussion on Nissl: https://forum.image.sc/t/automated-cell-counting/32770/4
*Edit: I dug out some of my Nissl test stains and tried the StarDist 2D Plugin (Fiji) using the pre-trained Model "Versatile (H&E)". The results look pretty good! In a second step, you should be able to filter out the neurons via the cell size, they should be bigger than the rest. Best of luck!
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