Life is dynamic and three-dimensional (3D). An ideal tool to study biological processes, particularly those involving large populations of functionally diverse cells and fast dynamics, such as brains, is to achieve volumetric imaging in vivo. Traditional imaging tools, such as point scanning confocal microscope, are too slow to catch rapid processes over large scales. Volumetric imaging techniques that completely parallelized the imaging collection start attracting increasingly more attentions recently. Here, we report several new types of light field microscopes that can do functional imaging of calcium activities over large populations of neurons at high speed. We demonstrated whole brain functional imaging in freely behaving larval zebrafish and captured activation of several functional neural ensembles during different phases of larval zebrafish prey capture behavior. We also recorded populations of neurons’ activity and track fast circulating blood cells in 3D vascular network in the mouse brain. We believe these volumetric high speed imaging techniques could open new windows to understand more dynamic biological processes in vivo.
The optical heterogeneity of biological tissue imposes a major limitation to acquire detailed structural and functional information deep in the biological specimens using conventional microscopes. To restore optimal imaging performance, we developed an adaptive optical microscope based on direct wavefront sensing technique. This microscope can reliably measure and correct biological samples induced aberration. We demonstrated its performance and application in structural and functional brain imaging in various animal models, including fruit fly, zebrafish and mouse.
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