Full view observation throughout entire specimens over a prolonged period is crucial when exploring the physiological functions and system-level behaviors.Multi-photon microscopy(MPM)has been widely employed for such ...Full view observation throughout entire specimens over a prolonged period is crucial when exploring the physiological functions and system-level behaviors.Multi-photon microscopy(MPM)has been widely employed for such purposes owing to its deep penetration ability.However,the current MPM struggles with balancing the imaging depth and quality while avoiding photodamage for the exponential increasement of excitation power with the imaging depth.Here,we present a dual-objective two-photon microscope(Duo-2P),characterized by bidirectional two-photon excitation and fluorescence collection,for long-duration volumetric imaging of dense scattering samples.Duo-2P effectively doubles the imaging depth,reduces the total excitation energy by an order of magnitude for samples with a thickness five times the scattering length,and enhances the signal-to-noise ratio up to 1.4 times.Leveraging these advantages,we acquired volumetric images of a 380-μm suprachiasmatic nucleus slice for continuous 4-h recording at a rate of 1.67 s/volume,visualized the calcium activities over 4000 neurons,and uncovered their state-switching behavior.We conclude that Duo-2P provides an elegant and powerful means to overcome the fundamental depth limit while mitigating photodamages for deep tissue volumetric imaging.展开更多
Dear Editor,Myocardial infarction(MI)is one of the leading causes of cardiovascular-related mortality worldwide.Timely restoration of the blood supply to ischemic my-ocardium by thrombolysis or percuta-neous coronary ...Dear Editor,Myocardial infarction(MI)is one of the leading causes of cardiovascular-related mortality worldwide.Timely restoration of the blood supply to ischemic my-ocardium by thrombolysis or percuta-neous coronary intervention is a com-mon clinical practice and decreases the mortality risk for MI patients(Heusch,2020).展开更多
An ultimate goal of neuroscience is to decipher the principles underlying neuronal information processing at the molecular,cellular,circuit,and system levels.The advent of miniature fluorescence microscopy has further...An ultimate goal of neuroscience is to decipher the principles underlying neuronal information processing at the molecular,cellular,circuit,and system levels.The advent of miniature fluorescence microscopy has furthered the quest by visualizing brain activities and structural dynamics in animals engaged in self-determined behaviors.In this brief review,we summarize recent advances in miniature fluorescence microscopy for neuroscience,focusing mostly on two mainstream solutions-miniature single-photon microscopy,and miniature two-photon microscopy.We discuss their technical advantages and limitations as well as unmet challenges for future improvement.Examples of preliminary applications are also presented to reflect on a new trend of brain imaging in experimental paradigms involving body movements,long and complex protocols,and even disease progression and aging.展开更多
DearEditor,Due to the tight and precise associations between behaviors and neural activities,the investigation of neuronal activity in freely-movingand behavinganimals isamajor unaccomplished goal in neuroscience.Epil...DearEditor,Due to the tight and precise associations between behaviors and neural activities,the investigation of neuronal activity in freely-movingand behavinganimals isamajor unaccomplished goal in neuroscience.Epileptiform seizures,characterized by hyper-synchronized discharges and hyper-excitation in neuronal networks,are one of the most common neurological disorders.展开更多
Ischemia-reperfusion injury occurs after reperfusion treatment for patients suffering myocardial infarction,however the underlying mechanisms are incompletely understood and effective pharmacological interventions are...Ischemia-reperfusion injury occurs after reperfusion treatment for patients suffering myocardial infarction,however the underlying mechanisms are incompletely understood and effective pharmacological interventions are limited.Here,we report the identification and characterization of the FDA-approved drug disulfiram(DSF)as a cardioprotective compound.By applying high-throughput chemical screening,we found that DSF decreased H_(2)O_(2)-induced cardiomyocyte death by inhibiting Gasdermin D,but not ALDH1,in cardiomyocytes.Oral gavage of DSF decreased myocardial infarct size and improved heart function after myocardial ischemia-reperfusion injury in rats.Therefore,this work reveals DSF as a potential therapeutic compound for the treatment of ischemic heart disease.展开更多
基金National Natural Science Foundation of China(32293210,32327802)CAMS Innovation Fund for Medical Sciences(2019-I2M-5-054).
文摘Full view observation throughout entire specimens over a prolonged period is crucial when exploring the physiological functions and system-level behaviors.Multi-photon microscopy(MPM)has been widely employed for such purposes owing to its deep penetration ability.However,the current MPM struggles with balancing the imaging depth and quality while avoiding photodamage for the exponential increasement of excitation power with the imaging depth.Here,we present a dual-objective two-photon microscope(Duo-2P),characterized by bidirectional two-photon excitation and fluorescence collection,for long-duration volumetric imaging of dense scattering samples.Duo-2P effectively doubles the imaging depth,reduces the total excitation energy by an order of magnitude for samples with a thickness five times the scattering length,and enhances the signal-to-noise ratio up to 1.4 times.Leveraging these advantages,we acquired volumetric images of a 380-μm suprachiasmatic nucleus slice for continuous 4-h recording at a rate of 1.67 s/volume,visualized the calcium activities over 4000 neurons,and uncovered their state-switching behavior.We conclude that Duo-2P provides an elegant and powerful means to overcome the fundamental depth limit while mitigating photodamages for deep tissue volumetric imaging.
基金supported by grants from the National Key R&D Program of China(2018YFA0800501,2019YFA0801602)the National Natural Science Foundation of China(32230032,31730061,31430059,81870198)and Synogen Biopharma Co.,Nanjing,China.
文摘Dear Editor,Myocardial infarction(MI)is one of the leading causes of cardiovascular-related mortality worldwide.Timely restoration of the blood supply to ischemic my-ocardium by thrombolysis or percuta-neous coronary intervention is a com-mon clinical practice and decreases the mortality risk for MI patients(Heusch,2020).
基金We thank Dr.Zhe Zhao and Dr.Haitao Wu for helping with the experiments for Fig.2D,and Dr.Weijian Zong for discussion.This work was supported by grants from the National Natural Science Foundation of China(31327901,31570839,61975002,31830036,31821091,and 8182780030)the Major State Basic Research Program of China(2016 YFA0500400 and 2016YFA0500403)and the National Postdoctoral Program for Innovative Talents of China(BX20190011).
文摘An ultimate goal of neuroscience is to decipher the principles underlying neuronal information processing at the molecular,cellular,circuit,and system levels.The advent of miniature fluorescence microscopy has furthered the quest by visualizing brain activities and structural dynamics in animals engaged in self-determined behaviors.In this brief review,we summarize recent advances in miniature fluorescence microscopy for neuroscience,focusing mostly on two mainstream solutions-miniature single-photon microscopy,and miniature two-photon microscopy.We discuss their technical advantages and limitations as well as unmet challenges for future improvement.Examples of preliminary applications are also presented to reflect on a new trend of brain imaging in experimental paradigms involving body movements,long and complex protocols,and even disease progression and aging.
基金This work was supported by the National Natural Science Foundation of China(91642205,81830038,91949208,and 82071327)the Natural Science Foundation of Tianjin Province(18JCYBJC43800)。
文摘DearEditor,Due to the tight and precise associations between behaviors and neural activities,the investigation of neuronal activity in freely-movingand behavinganimals isamajor unaccomplished goal in neuroscience.Epileptiform seizures,characterized by hyper-synchronized discharges and hyper-excitation in neuronal networks,are one of the most common neurological disorders.
基金supported by grants from the National Key R&D Program of China(2018YFA0800501 and 2019YFA0801602)the National Natural Science Foundation of China(32230032,31730061,31430059,and 81870198)Shandong Provincial Natural Science Foundation(ZR2022QH394).
文摘Ischemia-reperfusion injury occurs after reperfusion treatment for patients suffering myocardial infarction,however the underlying mechanisms are incompletely understood and effective pharmacological interventions are limited.Here,we report the identification and characterization of the FDA-approved drug disulfiram(DSF)as a cardioprotective compound.By applying high-throughput chemical screening,we found that DSF decreased H_(2)O_(2)-induced cardiomyocyte death by inhibiting Gasdermin D,but not ALDH1,in cardiomyocytes.Oral gavage of DSF decreased myocardial infarct size and improved heart function after myocardial ischemia-reperfusion injury in rats.Therefore,this work reveals DSF as a potential therapeutic compound for the treatment of ischemic heart disease.