I stained cryo-slides with GFAP, i had a good signal and i need a suggest for a good quantification, i have used the Integrated-density but i am not sure, if it was a good idea because the areas of DG in the stained slides are different!
You can do a morphometric study by uploading the photomicrographs of your slides unto the image J software. For morphometric analysis you would need serial sections and multiple fields. If you choose to use the automated counter you would need to threshold for colour or you could use the manual counter after applying a grid over the image. The astrocytes or immature granule cell usually stain positive for GFAP, while mature granule cells and the pyramidal cells usually from the cornus ammonis 4 region do not. comparisons against a normal control would help if you do not have a reference for number of cells in the region.
This articlecould also help: Neural Plast. 2016; 2016: 2426413.
Published online 2016 Aug 7. doi: 10.1155/2016/2426413
PMCID: PMC4992534
Anorexia Reduces GFAP+ Cell Density in the Rat Hippocampus
Daniel Reyes-Haro, * Francisco Emmanuel Labrada-Moncada, Durairaj Ragu Varman, Janina Krüger, Teresa Morales, Ricardo Miledi, and Ataúlfo Martínez-Torres
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