Scanless volumetric imaging by selective access multifocal multiphoton microscopy
Author(s)Xue, Yi; Berry, Kalen Paul; Boivin, Josiah R.; Rowlands, Christopher; Takiguchi, Yu; Nedivi, Elly; So, Peter T. C.; ... Show more Show less
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Simultaneous, high-resolution imaging across a large number of synaptic and dendritic sites is critical for understanding how neurons receive and integrate signals. Yet, functional imaging that targets a large number of submicrometer-sized synaptic and dendritic locations poses significant technical challenges. We demonstrate a new parallelized approach to address such questions, increasing the signal-to-noise ratio by an order of magnitude compared to previous approaches. This selective access multifocal multiphoton microscopy uses a spatial light modulator to generate multifocal excitation in three dimensions (3D) and a Gaussian–Laguerre phase plate to simultaneously detect fluorescence from these spots throughout the volume. We test the performance of this system by simultaneously recording Ca 2 dynamics from cultured neurons at 98–118 locations distributed throughout a 3D volume. This is the first demonstration of 3D imaging in a “single shot” and permits synchronized monitoring of signal propagation across multiple different dendrites.
DepartmentMassachusetts Institute of Technology. Department of Mechanical Engineering; Massachusetts Institute of Technology. Laser Biomedical Research Center; Massachusetts Institute of Technology. Department of Biology; Picower Institute for Learning and Memory; Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
The Optical Society
Xue, Yi et al. "Scanless volumetric imaging by selective access multifocal multiphoton microscopy." Optica 6, 1 (January 2019): 76-83 © 2019 Optical Society of America.
Final published version