Diolistic labeling and analysis of dendritic spines

Dendritic spines are diverse and plastic components of the neuronal cell apparatus and are highly responsive to trophic factors during both development and adulthood. Diolistic labeling of neurons with lipophilic fluorescent dyes, coupled with advanced high-resolution microscopy methods, provides robust labeling of dendritic spines for assessment of their density and morphology. Here, we describe a method for labeling of dendritic spines using diolistic labeling in ex vivo brain slices, visualization using confocal laser scanning microscopy, deconvolution, and analysis using the Surpass module of Bitplane Imaris software.