This technology is a protocol for generating 3D images of Purkinje Neurons across different layers of the cerebellum, at single cell resolution.
Purkinje cells are large neurons found in the brain, which are critical in controlling coordinated movement. Disruption of their function is therefore associated with a variety of neurodegenerative and neurodevelopmental diseases. Current methods of imaging these cells use immunofluorescence staining of a calcium binding protein. However, due to the high expression level of this protein in these cells, and their tightly layered distribution in the cerebellum, this method is not sufficient for resolving individual cells or 3D reconstruction.
This protocol uses immunofluorescence co-staining to collect high resolution images of a Purkinje neuron across multiple cerebellum layers. One stain highlights the cell bodies, while the other improves resolution of the dendrites. These images can then be stitched together to form 3D reconstructions of the full, individual cells. These images can reveal subtle cell defects, or loss of cells, as is seen in diseased brain tissue.
This technology has been validated in a murine model.
IR CU22200
Licensing Contact: Jerry Kokoshka
