Staining Neurons and counting them using CLARITY?

we are using the CLARITY technique to clear Rat brains and we would like to label astrocytes and neurons in the cerebral cortex and to be able to count them. We tried to stain it using NeuN and GFAP, but it appears that it didn’t stain when we did imaging. Do you have any recommendations/thoughts on why it did not work, or any specific steps we should follow?
we followed steps on
http://wiki.claritytechniques.org/index.php/Immunostaining

Comments

  • aysegulaysegul Posts: 33
    Hi maryam_z, how many days did you incubate your section in primer antibody and how much antibody dilution did you use?
  • maryam_zmaryam_z Posts: 2
    Hello aysegul. We incubated a 200 micrometer slides for 1 day and 2 days in 1:50 concentration primary antibody. We incubated them in 2 different temp: one at 37 degrees, and the other at 4 degrees. The same time and temp was repeated for secondaries antibodies.
    GFAP did stain, but NeuN didn't in any of the scenarios.
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