SBIR-STTR Award

This project aims to develop and commercialize the Volumetric Calcium Imaging 2-Photon Activity Microscope,vCAm™, a revolutionary new 2-photon microscope based on a technological breakthrough called Light BeadsMicroscopy (LBM) that was recently developed by Dr. Alipasha Vaziri and co-workers (Lab. Neurotechnol.Biophys., Rockefeller Univ., New York, NY). The game-changing innovation in the vCAm is the ability to performunparalleled in vivo calcium imaging of individual neurons at cellular resolution nearly simultaneously in one ormore cytoarchitectonic regions of the mouse cerebral cortex, and nearly simultaneously in 30 imaging planeseach ~16 µm apart (i.e., up to a total depth of 500 µm, encompassing layers I-V) at a full-frame rate of at least12 Hertz. These capabilities are crucial for ultimately correlating stimuli and/or behavioral states of an animaldiscretely, in a context-dependent manner, with the activity of all neurons in the brain of the animal that areinvolved in this process, which requires simultaneous recording of the activity of hundreds of thousands ofneurons in a multi-regional and multi-layer manner. However, contemporary 2-photon microscopy suffers froma fundamental limitation. Neuroscience researchers need to record simultaneous interactions between thesensory, motor and visual regions of the brain, but it is difficult to capture the activity in such a broad volume ofthe brain without sacrificing resolution or speed. The LBM technology pushes the limits of imaging speed to thephysical nature of fluorescence itself by eliminating the "dead time" between sequential laser pulses when noneuroactivity is recorded and at the same time the need for scanning. With this approach, the only limit to therate at which samples can be recorded is the time that it takes the tags to fluoresce, meaning wide volumes ofthe brain can be recorded within the same time it would take a conventional two-photon microscope to capturea much smaller number of brain cells. Other technology, such as miniaturized 2-photon microscopes that can becarried on the head of freely moving rodents, functional magnetic resonance imaging, inserting electrodes intothe brain, or fiber photometry do not fulfill this need. This project will improve upon the original LBM invention tocreate a commercial product for disseminating this important new technology. Based on pilot work performed atDr. Vaziri's laboratory, it is clear that the vCAm will make a significant impact on the field of neuroscienceresearch, including advancing studies focused on alterations in the circuitry of the central nervous systemassociated with neurodevelopmental, neuropsychiatric and neurodegenerative disorders. Ultimately, this willresult in an improved basis for developing novel treatment strategies for a wide spectrum of complex braindiseases. In Phase I we will demonstrate the feasibility of this novel technology by developing prototype hardwareand software; work in Phase II will focus on creating the full functionality of the vCAm for commercial release.We will perform extensive feasibility studies, product validation and usability studies of the vCAm in closecollaboration with Dr. Vaziri. A competing technology is not commercially available.

Public Health Relevance Statement:
Narrative This project develops a revolutionary new two-photon laser scanning microscope system based on a technological breakthrough called Light Beads Microscopy. This microscope will enable unparalleled in vivo calcium imaging of individual neurons nearly simultaneously in one or more regions of the mouse cerebral cortex, only limited by the rate that it takes the tags to fluoresce, meaning wide volumes of the brain can be recorded within the same time it would take a conventional two-photon microscope to capture a much smaller number of neurons. As a result, new avenues of neuroscience research will become possible providing promise for the elucidation of novel biological mechanisms associated with human brain disease, and ultimately the basis for developing novel therapies to prevent and treat complex brain diseases.

Project Terms:
<3-D><3D><3-Dimensional><2-photon><2-photon microscopy>

This project aims to develop and commercialize the Volumetric Calcium Imaging 2-Photon Activity Microscope,vCAm™, a revolutionary new 2-photon microscope based on a technological breakthrough called Light BeadsMicroscopy (LBM) that was recently developed by Dr. Alipasha Vaziri and co-workers (Lab. Neurotechnol.Biophys., Rockefeller Univ., New York, NY). The game-changing innovation in the vCAm is the ability to performunparalleled in vivo calcium imaging of individual neurons at cellular resolution nearly simultaneously in one ormore cytoarchitectonic regions of the mouse cerebral cortex, and nearly simultaneously in 30 imaging planeseach ~16 µm apart (i.e., up to a total depth of 500 µm, encompassing layers I-V) at a full-frame rate of at least12 Hertz. These capabilities are crucial for ultimately correlating stimuli and/or behavioral states of an animaldiscretely, in a context-dependent manner, with the activity of all neurons in the brain of the animal that areinvolved in this process, which requires simultaneous recording of the activity of hundreds of thousands ofneurons in a multi-regional and multi-layer manner. However, contemporary 2-photon microscopy suffers froma fundamental limitation. Neuroscience researchers need to record simultaneous interactions between thesensory, motor and visual regions of the brain, but it is difficult to capture the activity in such a broad volume ofthe brain without sacrificing resolution or speed. The LBM technology pushes the limits of imaging speed to thephysical nature of fluorescence itself by eliminating the "dead time" between sequential laser pulses when noneuroactivity is recorded and at the same time the need for scanning. With this approach, the only limit to therate at which samples can be recorded is the time that it takes the tags to fluoresce, meaning wide volumes ofthe brain can be recorded within the same time it would take a conventional two-photon microscope to capturea much smaller number of brain cells. Other technology, such as miniaturized 2-photon microscopes that can becarried on the head of freely moving rodents, functional magnetic resonance imaging, inserting electrodes intothe brain, or fiber photometry do not fulfill this need. This project will improve upon the original LBM invention tocreate a commercial product for disseminating this important new technology. Based on pilot work performed atDr. Vaziri's laboratory, it is clear that the vCAm will make a significant impact on the field of neuroscienceresearch, including advancing studies focused on alterations in the circuitry of the central nervous systemassociated with neurodevelopmental, neuropsychiatric and neurodegenerative disorders. Ultimately, this willresult in an improved basis for developing novel treatment strategies for a wide spectrum of complex braindiseases. In Phase I we will demonstrate the feasibility of this novel technology by developing prototype hardwareand software; work in Phase II will focus on creating the full functionality of the vCAm for commercial release.We will perform extensive feasibility studies, product validation and usability studies of the vCAm in closecollaboration with Dr. Vaziri. A competing technology is not commercially available.

Public Health Relevance Statement:
Narrative This project develops a revolutionary new two-photon laser scanning microscope system based on a technological breakthrough called Light Beads Microscopy. This microscope will enable unparalleled in vivo calcium imaging of individual neurons nearly simultaneously in one or more regions of the mouse cerebral cortex, only limited by the rate that it takes the tags to fluoresce, meaning wide volumes of the brain can be recorded within the same time it would take a conventional two-photon microscope to capture a much smaller number of neurons. As a result, new avenues of neuroscience research will become possible providing promise for the elucidation of novel biological mechanisms associated with human brain disease, and ultimately the basis for developing novel therapies to prevent and treat complex brain diseases.

Project Terms: