Experiments

Morphofunctional imaging of human and mouse brain

Neuronal imaging research team aims to develop innovative imaging methodologies for an increased understanding of biological events in human and mouse brain. Novel implementations of light-sheet microscopy are applied to resolve neuronal anatomy in whole fixed brains with cellular resolution. We combined the advantages of light-sheet illumination and confocal slit detection to increase the image contrast in real time. We find that confocal light-sheet allows reconstructing macroscopic brain volumes with sub-cellular resolution. We obtained a comprehensive map of whole fluorescently labeled mouse brains and macroscopic immunolabeled dysplastic human samples. Moving to living samples, real-time dynamics of brain rewiring are visualized through two-photon microscopy with the spatial resolution of single synaptic contacts. The plasticity of the injured brain is also dissected through cutting-edge optical methods capable to ablate a single axon or to produce a model of ischemic stroke. Further, to obtain a more comprehensive view of brain functionality, complementary approaches are combined into a correlative framework. Data from in vivo two-photon fluorescence imaging and ex vivo light sheet microscopy are integrated taking advantage of blood vessels as reference chart. The correlative approach can be applied to different mouse models presenting variously labeled fluorescent cells. Studies in which synaptic plasticity is monitored in vivo using two-photon fluorescence microscopy could benefit from the presented method to reconstruct the whole neuroanatomical features of the observed neuron.


Research & Technical staff:
Sacconi LeonardoSilvestri LudovicoFerrantini IreneCostantini Irene

Associated Researchers:
Pavone Francesco Saverio

English