8th Apr, 2017
King Abdulaziz University
Question
Asked 5th Apr, 2017
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 didn’t stain. Do you have any recommendations/thoughts on why it did not work, or any specific steps we should follow? we followed steps on
All Answers (3)
Clarity is perfect depending on your goal as clarity mapping preserves fine structure, proteins and nucleic acids, please follow this link
I suggest you only need to repeat your steps again once or twice for NeuN and GFAP markers.
Dear Maryam
If you still need help on this subject it would be helpful to provide more information, such as the following:
How thick is your tissue? How long have you cleared it? Did you use PACT (A4P0)? What were your staining conditions (antibody, concentrations, time, temperature)?
What is your imaging setup?
Otherwise, I recommend the article below, where the authors stained and counted PV-neurons in the cortex. They provide several suggestions to improve staining efficiency in thick CLARITY tissue.
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