To understand how the brain computes, we need to understand how individual neurons in a circuit integrate their numerous inputs into output signals, as well as how they work together to encode a sensory input or execute a motor command in a behaving animal. Circuits and neurons are three-dimensional (3D) and can extend over hundreds or thousands of microns. Therefore, understanding their operations requires monitoring their activity at both synaptic and cellular resolution in 3D at image rates that capture all activity events. Behaving animals present a host of challenges to this goal. Existing 3D imaging technologies suffer from insufficient volume imaging speed, brain-motion-induced image artifacts, as well as complex hardware and software implementation. These limitations have prevented their adoption by biology laboratories and remain a technical barrier for neuroscience research. Successful completion of our proposal will overcome these limitations and profoundly impact neuroscience research. We recently developed a Bessel focus scanning technology (BEST) that is easily integrated into existing two-photon microscopes, resistant to motion artifacts, and have already achieved 30-Hz, synapse-resolving volumetric imaging of sparsely labelled neuronal populations in a wide variety of model organisms. In this proposal, combining the expertise of microscopists, biologists, and data scientists, we propose to further optimize BEST to enable high-speed, high-throughput, and high-resolution volumetric activity recording of both sparsely and densely labelled circuits throughout the living brain.
We aim to record whole-brain activity in the fly at >10 Hz and through-cortex volume imaging in the mouse at ~2Hz. By combining BEST with microendoscopy, we aim to achieve synaptic-resolution volumetric microendoscopic imaging at 30 Hz and use it to study structural and functional plasticity in deeply buried nuclei of the mouse brain. By correcting brain-induced optical aberrations, adaptive optics will enable BEST to maintain synapse resolution throughout the entire mouse cortex. Easily adoptable, BEST has already been integrated into multiple two-photon fluorescence microscopes in laboratories worldwide. With a continuously expanding user base, the proposed optimization project will immediately benefit a wide range of laboratories, allowing them to study volumetric neural activity at unprecedented high spatiotemporal resolution throughout the living brain.

Public Health Relevance

To develop cures to neurological diseases, we must have a mechanistic understanding of neural circuit functions in the brain, which requires the ability to monitor neural activity in three dimensions at high spatial and temporal resolution. Leveraging recent advances in imaging technologies and data sciences, our proposal will enable, for the first time, high-speed, high-resolution, and high-throughput volumetric imaging of neural circuit activity throughout the entire living brain.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project--Cooperative Agreements (U01)
Project #
1U01NS103489-01
Application #
9404832
Study Section
Special Emphasis Panel (ZNS1)
Program Officer
Talley, Edmund M
Project Start
2017-07-15
Project End
2020-06-30
Budget Start
2017-07-15
Budget End
2018-06-30
Support Year
1
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of California Berkeley
Department
Physics
Type
Schools of Arts and Sciences
DUNS #
124726725
City
Berkeley
State
CA
Country
United States
Zip Code
94704
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Rodríguez, Cristina; Ji, Na (2018) Adaptive optical microscopy for neurobiology. Curr Opin Neurobiol 50:83-91
Rodríguez, Cristina; Liang, Yajie; Lu, Rongwen et al. (2018) Three-photon fluorescence microscopy with an axially elongated Bessel focus. Opt Lett 43:1914-1917
Lu, Rongwen; Tanimoto, Masashi; Koyama, Minoru et al. (2018) 50 Hz volumetric functional imaging with continuously adjustable depth of focus. Biomed Opt Express 9:1964-1976