Abstract - Joel Kubby
We propose to develop a three-photon excitation (3PE) microscope with adaptive optics (AO) for deep brain imaging in live animals. Brain structures such as hippocampus and striatum play essential roles in navigation, decision making, learning and memory, but reside at a depth inaccessible to current optical microscopy. Our proposed microscope will be based on an Olympus FV1200MPE two-photon microscope, supplemented with a low duty cycle (10 MHz), high pulse energy (50 nJ) femto-second (80 fs) fiber laser for 3PE, and an AO system for wavefront sensing and correction. The long wavelength (1.55μm) 3PE will significantly decrease scattering, hence increasing the imaging depth and signal-to-background ratio. AO will correct aberrations due to inhomogeneous refractive indices in brain tissue to achieve better resolution. The 3PE laser will be suitable for common fluorescent proteins with excitation peaks near 517nm (~1.55μm/3), such as EYFP, Venus, and mCitrine. The microscope will enable anatomical and functional imaging of neural circuits with sub-cellular resolution 1,500-1,800μm below the brain surface, 3-5 times deeper than commonly achieved with two-photon microscopy. It will greatly facilitate research on structural alterations (e.g., dendritic spine dynamics) and neuronal activities (e.g., Ca signals) in deep cortical layers and subcortical regions including hippocampus and striatum.
AWARDS
| Principal Investigator | Institution | Title | Abstract |
| Andersen, Richard | California Institute of Technology | Engineering Artificial Sensation | View |
| Andrews, Anne | University of California, Los Angeles | Nanoscale Neurotransmitter Sensors | View |
| Bloodgood, Brenda | University of California San Diego | A novel toolkit for visualizing and manipulating activity-induced transcription in living brain. | View |
| Chaumeil, Myriam | University of California, San Francisco | In vivo metabolic imaging of neuroinflammation using hyperpolarized 13C | View |
| Cleary, Michael | University of California, Merced | Capturing physiological maps of neural gene expression | View |
| Cohen, Bruce | University of California, Lawrence Berkeley National Laboratory | Nano-optogenetic control of neuronal firing with targeted nanocrystals | View |
| Dai, Hongjie | Stanford University | Deep brain imaging of single neurons in the second near-infrared optical window | View |
| Hall, Drew | University of California, San Diego | Magnetic Monitoring of Neural Activity using Magnetoresistive Nanosensors | View |
| Krubitzer, Leah | University of California, Davis | An integrated system to monitor, image, and regulate neural activity | View |
| Kubby, Joel | University of California, Santa Cruz | Three-Photon Microscopy with Adaptive Optics for Deep Tissue Brain Activity Imaging | View |
| Melosh, Nicholas | Stanford University | Parallel Solid State Intracellular Patch-Clamping with Biomimetic Probes | View |
| Park, B. Hyle | University of California, Riverside | Label-free 4D optical detection of neural activity | View |
| Portera-Cailliau, Carlos | University of California, Los Angeles | High-speed interrogation of network activity with frequency domain multiplexing | View |
| Shanechi, Maryam | University of Southern California | Control-Theoretic Neuroprosthetic Design Using Electrocorticography Signals | View |
| Smith, Will | University of California, Santa Barbara | Whole brain imaging in a primative chordate | View |
| Wood, Marcelo | University of California, Irvine | Epigenetic PET tracer for cross-species investigation of age-related memory dysfunction | View |