Techniques / Tools
In vivo imaging of layer 5 pyramidal neurons in the somatosensory cortex (labeled using the Thy1-GFP mouse
We are studying the role of immune genes in cortical synaptic development. During early postnatal development, some synapses are eliminated, in part, via microglia-dependent synaptic phagocytosis. We use a method called Expansion microscopy (ExM, from Ed Boyden's group) to visualize this biological process.
We can visualize synapses using FingRs (Fibronectin intrabodies generated with mRNA display), that bind endogenous neuronal proteins PSD-95 (Gross et al. 2013). Below are labeled synapses in a dendrite (Comer et al. 2020).
We can identify different neuronal subtypes using multiplex in situ hybridization (M-FISH, RNAScope). Here we stain for parvalbumin and somatostatin in the prefrontal cortex of a mouse
We can dissect the connectivity patterns of moleculary identified brain cell types using simultaneous whole-cell patch-clmap recordings.
IMARIS reconstruction based on STED microscopy. Using this nano-resolution approach we identified PSD95 (green) and Rab11a (magenta) puncta that are within 0.2 um from the border of the spines.