We present a threedimensional(3D)isotropic imaging of mouse brain using light-sheet fuo-rescent microscopy(LSFM)in conjumction with a multi-view imaging computation.Unlike common single view LSFM is used for mouse bra...We present a threedimensional(3D)isotropic imaging of mouse brain using light-sheet fuo-rescent microscopy(LSFM)in conjumction with a multi-view imaging computation.Unlike common single view LSFM is used for mouse brain imaging,the brain tissue is 3D imaged under eight views in our study,by a home-built selective plane ilumination microscopy(SPIM).An output image containing complete structural infornation as well as significantly improved res olution(~4 times)are then computed based on these eight views of data,using a bead-guided multi-view registration and deconvolution.With superior imaging quality,the astrocyte and pyrarmidal neurons together with their subcellular nerve fbers can be clearly visualized and segmented.With further incuding other computational methods,this study can be potentially scaled up to map the conectome of whole mouse brain with a simple light.sheet microscope.展开更多
Bronchus-associated lymphoid tissue(BALT)develops at unpredictable locations around lung bronchi following pulmonary inflammation.The formation and composition of BALT have primarily been investigated by immunohistolo...Bronchus-associated lymphoid tissue(BALT)develops at unpredictable locations around lung bronchi following pulmonary inflammation.The formation and composition of BALT have primarily been investigated by immunohistology that,due to the size of the invested organ,is usually restricted to a limited number of histological sections.To assess the entire BALT of the lung,other approaches are urgently needed.Here,we introduce a novel light sheet microscopy-based approach for assessing lymphoid tissue in the lung.Using antibody staining of whole lung lobes and optical clearing by organic solvents,we present a method that allows in-depth visualization of the entire bronchial tree,the lymphatic vasculature and the immune cell composition of the induced BALT.Furthermore,three-dimensional analysis of the entire lung allows the qualitative and quantitative enumeration of the induced BALT.Using this approach,we show that a single intranasal application of the replication-deficient poxvirus MVA induces BALT that constitutes up to 8%of the entire lung volume in mice deficient in CCR7,in contrast to wild type mice(WT).Furthermore,BALT induced by heat-inactivated E.coli is dominated by a pronounced T cell infiltration in Cxcr5-deficient mice,in contrast to WT mice.展开更多
Understanding brain structure and function,and the complex relationships bet ween them,is one of the grand challenges of contemporary sciences.Thanks to their fexiblity,optical techniques could be the key to explore t...Understanding brain structure and function,and the complex relationships bet ween them,is one of the grand challenges of contemporary sciences.Thanks to their fexiblity,optical techniques could be the key to explore this complex network.In this manuscript,we briefly review recent adv ancements in optical methods applied to three main issues:anatomy,plasticity and func-tionality.We describe novel implement ations of light-sheet microscopy to resolve neuronal anat-omy in whole fixed brains with cellular resolution.Moving to liv ing samples,we show how real-time dynamics of brain rewiring can be visualized through two-photon microscopy with the spatial resolution of single synaptic contacts.The plasticity of the injured brain can also be dissected through cut ting edge optical methods that specifically ablate single neuronal processes.Finally,we report how nonlinear microscopy in combination with novel voltage sensitive dyes allow optical registrations of action potential across a population of neurons opening promising prospective in understanding brain functionality.The knowledge acquired from these complementary optical methods may provide a deeper comprehension of the brain and of its unique features.展开更多
基金funding support from 1000 Youth Talents Plan of China (P.F.)Fundamental Research Program of Shenzhen (P.F.,JCYJ20160429182424047)+1 种基金National Science Foundation of China (NSFC31571002,D.Z)Graduates'Innovation Fund of Huazhong University of Science and Technology (5003182004).
文摘We present a threedimensional(3D)isotropic imaging of mouse brain using light-sheet fuo-rescent microscopy(LSFM)in conjumction with a multi-view imaging computation.Unlike common single view LSFM is used for mouse brain imaging,the brain tissue is 3D imaged under eight views in our study,by a home-built selective plane ilumination microscopy(SPIM).An output image containing complete structural infornation as well as significantly improved res olution(~4 times)are then computed based on these eight views of data,using a bead-guided multi-view registration and deconvolution.With superior imaging quality,the astrocyte and pyrarmidal neurons together with their subcellular nerve fbers can be clearly visualized and segmented.With further incuding other computational methods,this study can be potentially scaled up to map the conectome of whole mouse brain with a simple light.sheet microscope.
基金This work was supported by a Lichtenberg-stipend to DTM through the University of Veterinary Medicine,Hannover,the German Excellence Initiative grant EXC62-Rebirth(to RF)DFG grant SFB 900/B1(to RF).
文摘Bronchus-associated lymphoid tissue(BALT)develops at unpredictable locations around lung bronchi following pulmonary inflammation.The formation and composition of BALT have primarily been investigated by immunohistology that,due to the size of the invested organ,is usually restricted to a limited number of histological sections.To assess the entire BALT of the lung,other approaches are urgently needed.Here,we introduce a novel light sheet microscopy-based approach for assessing lymphoid tissue in the lung.Using antibody staining of whole lung lobes and optical clearing by organic solvents,we present a method that allows in-depth visualization of the entire bronchial tree,the lymphatic vasculature and the immune cell composition of the induced BALT.Furthermore,three-dimensional analysis of the entire lung allows the qualitative and quantitative enumeration of the induced BALT.Using this approach,we show that a single intranasal application of the replication-deficient poxvirus MVA induces BALT that constitutes up to 8%of the entire lung volume in mice deficient in CCR7,in contrast to wild type mice(WT).Furthermore,BALT induced by heat-inactivated E.coli is dominated by a pronounced T cell infiltration in Cxcr5-deficient mice,in contrast to WT mice.
基金The research leading to these results has received funding from the European Union Seventh Frame-work Programme(FP7/2007-2013)under Grant agreements Nos.228334 and 241526This research project has also been supported by Human Frontier Science Program research grant(RGP0027/2009)the ItalianMinistry for Education,University and Research in the framework of the Flagship Pro-ject Nanomax and by Italian Ministry of Health in the framework of the"Stem cells call for proposals".This research has been carried out in the framework of the research activities of ICON foundation sup-ported by"Ente Cassa di Risparmio di Firenze".
文摘Understanding brain structure and function,and the complex relationships bet ween them,is one of the grand challenges of contemporary sciences.Thanks to their fexiblity,optical techniques could be the key to explore this complex network.In this manuscript,we briefly review recent adv ancements in optical methods applied to three main issues:anatomy,plasticity and func-tionality.We describe novel implement ations of light-sheet microscopy to resolve neuronal anat-omy in whole fixed brains with cellular resolution.Moving to liv ing samples,we show how real-time dynamics of brain rewiring can be visualized through two-photon microscopy with the spatial resolution of single synaptic contacts.The plasticity of the injured brain can also be dissected through cut ting edge optical methods that specifically ablate single neuronal processes.Finally,we report how nonlinear microscopy in combination with novel voltage sensitive dyes allow optical registrations of action potential across a population of neurons opening promising prospective in understanding brain functionality.The knowledge acquired from these complementary optical methods may provide a deeper comprehension of the brain and of its unique features.
