Monitoring large-scale neuronal activity in deep brain circuits is crucial for understanding the principles of brain function. However, existing large-scale cellular-resolution imaging techniques are limited in imaging depth and cannot monitor activity in areas deeper than the cortex in the intact brain. To address this challenge, we developed a Large Imaging Field of view Three-photon (LIFT) microscope with a FOV of > 3 mm2.
LIFT combines high-numerical-aperture optimized sampling, using a custom scanning module, with deep learning-based denoising, to enable population imaging in subcortical brain regions of mice and deep cortical regions of rats. We demonstrate calcium imaging in the intact mouse brain from more than 1500 cells across CA1, the surrounding white matter, and adjacent deep layers of the cortex, and show population imaging with high SNR at a depth of 1.2 mm in the rat cortex. The LIFT microscope offers versatility in imaging scale and rate and was built entirely from off-the-shelf components.
We would like to thank Dr. Anderson Chen (Neurophotonics Center, Boston University) for help with the mechanical design of LIFT scope, and Prof. David Boas (Boston University) for his advice and support.
The authors also thank Eric Lieser, Hongzhou Ma, and Jeff Brooker (Thorlabs Inc.) for helpful discussions, technical assistance, and providing critical parts of the microscope. Research reported in this publication was supported by awards R01 NS116139 and R56 MH132732 from the National Institutes of Health (NIH), by the Neurophotonics Center at Boston University, and by the Research Corporation for Science Advancement. A.M. is supported by the NIH DSPAN award, and H.F. is supported by the Israeli National Postdoctoral Award and by the American Committee for the Weizmann Institute of Science.
Department of Psychological and Brain Sciences, Boston University, Boston, MA, USA Hadas Frostig, Urvi Mishra, Siria Coello, Rebecca L. Suthard, Steve Ramirez & Benjamin B. Scott Graduate Program for Neuroscience, Boston University, Boston, MA, USA Department of Biology, Boston University, Boston, MA, USA Department of Biomedical Engineering, Boston University, Boston, MA, USA John T.
Giblin, Steve Ramirez, Jerome C. Scott Neurophotonics Center, Boston University, Boston, MA, USA Center for Systems Neuroscience, Boston University, Boston, MA, USA Steve Ramirez, Jerome C. Scott Photonics Center, Boston University, Boston, MA, USA Correspondence to Hadas Frostig, Jerome C.
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