Intravital Imaging and Analysis of the Structural Plasticity of Dendritic Spines in Multiple Brain Regions

The structural plasticity of dendritic spines is associated with changes in various cognitive performances across physiological and pathological conditions. With the rapid development of imaging and labelling methods, in vivo study of dendritic spine dynamics has been made possible from superficial to deep layers in many brain regions. However, depending on the brain region, there are multiple variants of surgical, imaging, and analyzing procedures, making them, as a whole, technically difficult to comprehend. Here, we introduce an open-skull coverslip-based surgical procedure of cranial window implantation, which is suitable for monitoring dendritic spine dynamics in the superficial cortical layer and can be easily adapted for monitoring brain regions of various depths and locations with minor modifications. Following these surgical procedures, we further describe intravital two-photon microscopy-based imaging approaches and methods for processing and analyzing acquired imaging data.