Photoacoustic(PA)microscopy is being increasingly used to visualize the microcirculation of the brain cortex at the micron level in living rodents.By combining it with long-term cranial window techniques,vasculature c...Photoacoustic(PA)microscopy is being increasingly used to visualize the microcirculation of the brain cortex at the micron level in living rodents.By combining it with long-term cranial window techniques,vasculature can be monitored over a period of days extending to months through a field of view.To fulfill the requirements of long-term in vivo PA imaging,the cranial window must involve a simple and rapid surgical procedure,biological compatibility,and sufficient optical-acoustic transparency,which are major challenges.Recently,several cranial window techniques have been reported for longitudinal PA imaging.Here,the development of chronic cranial windows for PA imaging is reviewed and its technical details are discussed,including window installation,imaging quality,and longitudinal stability.展开更多
Monitoring neuronal activity in vivo is critical to understanding the physiological or pathological functions of the brain.Two-photon Ca^(2+)imaging in vivo using a cranial window and specific neuronal labeling enable...Monitoring neuronal activity in vivo is critical to understanding the physiological or pathological functions of the brain.Two-photon Ca^(2+)imaging in vivo using a cranial window and specific neuronal labeling enables realtime,in situ,and long-term imaging of the living brain.Here,we constructed a recombinant rabies virus containing the Ca^(2+)indicator GCaMP6 s along with the fluorescent protein DsRed2 as a baseline reference to ensure GCaMP6 s signal reliability.This functional tracer was applied to retrogradely label specific V1-thalamus circuits and detect spontaneous Ca^(2+)activity in the dendrites of V1 corticothalamic neurons by in vivo two-photon Ca^(2+)imaging.Notably,we were able to record single-spine spontaneous Ca2+activity in specific circuits.Distinct spontaneous Ca^(2+)dynamics in dendrites of V1 corticothalamic neurons were found for different V1-thalamus circuits.Our method can be applied to monitor Ca^(2+)dynamics in specific input circuits in vivo,and contribute to functional studies of defined neural circuits and the dissection of functional circuit connections.展开更多
Imaging cells and microvasculature in the living brain is crucial to understanding an array of neurobiological phenomena.Here,we introduce a skull optical clearing window for imaging cortical structures at synaptic re...Imaging cells and microvasculature in the living brain is crucial to understanding an array of neurobiological phenomena.Here,we introduce a skull optical clearing window for imaging cortical structures at synaptic resolution.Combined with two-photon microscopy,this technique allowed us to repeatedly image neurons,microglia and microvasculature of mice.We applied it to study the plasticity of dendritic spines in critical periods and to visualize dendrites and microglia after laser ablation.Given its easy handling and safety,this method holds great promise for application in neuroscience research.展开更多
Bedside monitoring of brain function in severely brain-injured patients remains a critical clinical challenge.We demonstrate the translational potential of functional ultrasound(fUS)imaging for this purpose.In 6 comat...Bedside monitoring of brain function in severely brain-injured patients remains a critical clinical challenge.We demonstrate the translational potential of functional ultrasound(fUS)imaging for this purpose.In 6 comatose patients(Glasgow coma scale≤8)with cranial windows after decompressive craniectomy,we used a 7.8-MHz transducer optimized for cortical depths of 1.5 to 4 cm to perform real-time fUS during auditory stimulation.We observed task-related increases in regional cerebral blood flow(rCBF)in relevant brain regions(P<0.001,t test),which correlated with subsequent neurological recovery at 9-month follow-up.These findings establish fUS as a sensitive and portable tool for bedside brain function assessment,offering potential for improved prognostication,treatment guidance,and development of targeted rehabilitative strategies.展开更多
基金This study was supported by the National Natural Science Foundation of China,Nos.62022037,61775028,81571722,61528401Department of Science and Technology of Guangdong Province,Nos.2019ZT08Y191,SZBL2020090501013+1 种基金Shenzhen Science and Technology ProgramNos.KQTD20190929172743294,JCYJ20200109141222892Startup grant from Southern University of Science and Technology。
文摘Photoacoustic(PA)microscopy is being increasingly used to visualize the microcirculation of the brain cortex at the micron level in living rodents.By combining it with long-term cranial window techniques,vasculature can be monitored over a period of days extending to months through a field of view.To fulfill the requirements of long-term in vivo PA imaging,the cranial window must involve a simple and rapid surgical procedure,biological compatibility,and sufficient optical-acoustic transparency,which are major challenges.Recently,several cranial window techniques have been reported for longitudinal PA imaging.Here,the development of chronic cranial windows for PA imaging is reviewed and its technical details are discussed,including window installation,imaging quality,and longitudinal stability.
基金supported by the National Natural Science Foundation of China(31700934 and 31371106)。
文摘Monitoring neuronal activity in vivo is critical to understanding the physiological or pathological functions of the brain.Two-photon Ca^(2+)imaging in vivo using a cranial window and specific neuronal labeling enables realtime,in situ,and long-term imaging of the living brain.Here,we constructed a recombinant rabies virus containing the Ca^(2+)indicator GCaMP6 s along with the fluorescent protein DsRed2 as a baseline reference to ensure GCaMP6 s signal reliability.This functional tracer was applied to retrogradely label specific V1-thalamus circuits and detect spontaneous Ca^(2+)activity in the dendrites of V1 corticothalamic neurons by in vivo two-photon Ca^(2+)imaging.Notably,we were able to record single-spine spontaneous Ca2+activity in specific circuits.Distinct spontaneous Ca^(2+)dynamics in dendrites of V1 corticothalamic neurons were found for different V1-thalamus circuits.Our method can be applied to monitor Ca^(2+)dynamics in specific input circuits in vivo,and contribute to functional studies of defined neural circuits and the dissection of functional circuit connections.
基金supported by the National Natural Science Foundation of China(Grants Nos.91232710,31571002)the Science Fund for Creative Research Groups(Grant No.61721092)the Director Fund of WNLO.We are thankful to ZH Zhang for providing the Cx3cr1^(EGFP/+)mice.
文摘Imaging cells and microvasculature in the living brain is crucial to understanding an array of neurobiological phenomena.Here,we introduce a skull optical clearing window for imaging cortical structures at synaptic resolution.Combined with two-photon microscopy,this technique allowed us to repeatedly image neurons,microglia and microvasculature of mice.We applied it to study the plasticity of dendritic spines in critical periods and to visualize dendrites and microglia after laser ablation.Given its easy handling and safety,this method holds great promise for application in neuroscience research.
基金supported in part by the National Key Research and Development Program of China(grant no.2023YFC2410900)the National Natural Science Foundation of China(32371151)+4 种基金Guangdong High Level Innovation Research Institute(2021B0909050004)the Hong Kong Research Grants Council Collaborative Research Fund(C5053-22GF)General Research Fund(15224323 and 15104520)Hong Kong Innovation Technology Fund(MHP/014/19)internal funding from the Hong Kong Polytechnic University(G-SACD and 1-CDJM).
文摘Bedside monitoring of brain function in severely brain-injured patients remains a critical clinical challenge.We demonstrate the translational potential of functional ultrasound(fUS)imaging for this purpose.In 6 comatose patients(Glasgow coma scale≤8)with cranial windows after decompressive craniectomy,we used a 7.8-MHz transducer optimized for cortical depths of 1.5 to 4 cm to perform real-time fUS during auditory stimulation.We observed task-related increases in regional cerebral blood flow(rCBF)in relevant brain regions(P<0.001,t test),which correlated with subsequent neurological recovery at 9-month follow-up.These findings establish fUS as a sensitive and portable tool for bedside brain function assessment,offering potential for improved prognostication,treatment guidance,and development of targeted rehabilitative strategies.
