Synapse maps - confocal

We have processed brain material from transgenic mice expressing the enhanced green fluorescent protein (EGFP) coupled with the synaptic protein PSD95 (EGFP95) in order to accomplish the automated counting process using 3D connected components of fluorescent puncta in stacks of confocal microscope images for the CA1 region of the hippocampal formation. Furthermore, we have developed a PSDs volume segmentation and quantification algorithm designed to be executed in a supercomputer in order to obtain results in multiple brain regions in a short time. We have currently uploaded 6 examples of the 3D automated counting process of the PSD95 protein using confocal microscopy, from the following CA1 regions: stratum oriens (SO), CA1 stratum radiatum (SR), CA1 stratum lacunosum moleculare (SLM). In addition, brain sections in the same regions and layers as those proccesed for confocal microscopy, have been processed for 3D electron microscopy using FIB/SEM in order to crossvalidate and interpret confocal data with ultrastructural data with on the number and size of PSD puncta in different brain regions. We have currently uploaded 1 example of the 3D counting process of PSD puncta using FIB/SEM from the CA1 SR region.