SPED microscopy - high-speed, cellular-resolution volumetric imaging

Dr. Raju Tomer and Prof. Karl Deisseroth have developed “SPED” (Spherical-aberration-assisted Extended Depth-of-field) light sheet microscopy that combines a large volumetric field-of-view, via extended detection depth-of-field, with the optical sectioning of light sheet microscopy to enable high-speed functional and structural mapping of entire vertebrate nervous systems and CLARITY cleared intact brains at sub-cellular resolution. This system eliminates the need to physically scan the detection objective for volumetric imaging, therefore breaks a fundamental volumetric imaging speed barrier to enable potential scanning speeds of several thousand volumes per second. SPED-light sheet microscopy provides a fundamentally new approach for high-speed mapping of cellular activity in neural networks, with potentially broad applications for research and diagnostics.

The image shows an example 3D volume rendering from SPED high-speed functional imaging of an entire zebrafish larval nervous system, overlaid on SPED detection point spread functions across the field-of-view.

Stage of Research
The inventors have demonstrated the capabilities of SPED microscopy by performing sub-cellular resolution imaging of CLARITY mouse brains. They have also performed functional imaging of the entire zebrafish larval nervous system up to 12 volumes (40 planes) per second. Furthermore, they used their data to analyze neuronal activity spanning the entire naturally functioning nervous system.