Puncta Analyzer
The Puncta Analyzer protocol details how to quantify synapse number both in dissociated neuronal culture and in brain sections using immunocytochemistry. Using compartment-specific antibodies, we label presynaptic terminals as well as sites of postsynaptic specialization. We define synapses as points of colocalization between the signals generated by these markers. The number of these colocalizations is quantified using a plug in Puncta Analyzer (available upon request from cagla.eroglu@duke.edu under the ImageJ* analysis software platform. The synapse assay described in this protocol can be applied to any neural tissue or culture preparation for which you have selective pre- and postsynaptic markers. This synapse assay is a valuable tool that is already utilized in a number of studies on synaptic connectivity. (See the studies that used this method)
*Puncta Analyzer does not function with new versions of Image J. Please download and use ImageJ 1.29.
GEARBOCS
GEARBOCS is an Adeno Associated Virus Tool for In Vivo Gene Editing in Astrocytes. See preprint. The plasmids are available through addgene (#s 196495, 196489, 196490, 196491, 196492).