Parallelized fluorescence imaging has been a long-standing pursuit that can address the unmet need for a comprehensive three-dimensional(3D)visualization of dynamical biological processes with minimal photodamage.Howe...Parallelized fluorescence imaging has been a long-standing pursuit that can address the unmet need for a comprehensive three-dimensional(3D)visualization of dynamical biological processes with minimal photodamage.However,the available approaches are limited to incomplete parallelization in only two dimensions or sparse sampling in three dimensions.We hereby develop a novel fluorescence imaging approach,called coded light-sheet array microscopy(CLAM),which allows complete parallelized 3D imaging without mechanical scanning.Harnessing the concept of an“infinity mirror”,CLAM generates a light-sheet array with controllable sheet density and degree of coherence.Thus,CLAM circumvents the common complications of multiple coherent light-sheet generation in terms of dedicated wavefront engineering and mechanical dithering/scanning.Moreover,the encoding of multiplexed optical sections in CLAM allows the synchronous capture of all sectioned images within the imaged volume.We demonstrate the utility of CLAM in different imaging scenarios,including a light-scattering medium,an optically cleared tissue,and microparticles in fluidic flow.CLAM can maximize the signal-to-noise ratio and the spatial duty cycle,and also provides a further reduction in photobleaching compared to the major scanning-based 3D imaging systems.The flexible implementation of CLAM regarding both hardware and software ensures compatibility with any light-sheet imaging modality and could thus be instrumental in a multitude of areas in biological research.展开更多
基金financially supported by the Research Grants Council of the Hong Kong Special Administrative Region of China(HKU 17209017,17259316,17207715,C7047-16G,HKU 17209018,E-HKU701/17,CityU T42-103/16-N,and HKU 17205215)the Innovation and Technology Support Program(ITS/204/18 and GHP/024/16GD)+1 种基金the University Development Funds of the University of Hong KongNatural Science Foundation of China(N_HKU712/16).
文摘Parallelized fluorescence imaging has been a long-standing pursuit that can address the unmet need for a comprehensive three-dimensional(3D)visualization of dynamical biological processes with minimal photodamage.However,the available approaches are limited to incomplete parallelization in only two dimensions or sparse sampling in three dimensions.We hereby develop a novel fluorescence imaging approach,called coded light-sheet array microscopy(CLAM),which allows complete parallelized 3D imaging without mechanical scanning.Harnessing the concept of an“infinity mirror”,CLAM generates a light-sheet array with controllable sheet density and degree of coherence.Thus,CLAM circumvents the common complications of multiple coherent light-sheet generation in terms of dedicated wavefront engineering and mechanical dithering/scanning.Moreover,the encoding of multiplexed optical sections in CLAM allows the synchronous capture of all sectioned images within the imaged volume.We demonstrate the utility of CLAM in different imaging scenarios,including a light-scattering medium,an optically cleared tissue,and microparticles in fluidic flow.CLAM can maximize the signal-to-noise ratio and the spatial duty cycle,and also provides a further reduction in photobleaching compared to the major scanning-based 3D imaging systems.The flexible implementation of CLAM regarding both hardware and software ensures compatibility with any light-sheet imaging modality and could thus be instrumental in a multitude of areas in biological research.
