Membrane potential and its fluctuation are fundamental biophysical phenomena essential to cellular activities and functions.Compared to traditional electrode-based techniques,the optical recording via developed geneti...Membrane potential and its fluctuation are fundamental biophysical phenomena essential to cellular activities and functions.Compared to traditional electrode-based techniques,the optical recording via developed genetically encoded voltage indicators(GEVIs)offers a combination of noninvasiveness,high spatial resolution,and increased measurement throughput.However,its application is limited by the insufficient acquisition rate and time accuracy of the camera.Here we design and apply a stroboscopic illumination scheme to boost the temporal resolution of voltage imaging,while simultaneously eliminating the artifacts caused by nonsynchronized exposure in the rolling-shutter mode.We demonstrate that commonly used GEVIs are compatible with stroboscopic voltage imaging(SVI),and our SVI scheme offers a 5-fold faster acquisition frame rate than that of conventional continuous illumination.The GEVIs tested maintain high sensitivities in the SVI mode,supporting faithful reports of intracellular depolarization waveform and intercellular gap junction-mediated depolarization coupling in human embryonic kidney 293T(HEK 293T)cell populations.SVI allows resolving the action potential(AP)waveform with less distortion and mapping action potential initiation and propagation dynamics in cultured neurons in kilohertz,beyond the restriction from the camera in the field of view.展开更多
基金Prof.Zhixing Chen(Peking University)for providing 1,3,5,7-cyclooctatetraene-1-carboxylic acid(COT-COOH)supported by the Ministry of Science and Technology(2022YFA1304700,2018YFA0507600)the National Natural Science Foundation of China(32088101).P.Z.is sponsored by Bayer Investigator Award.
文摘Membrane potential and its fluctuation are fundamental biophysical phenomena essential to cellular activities and functions.Compared to traditional electrode-based techniques,the optical recording via developed genetically encoded voltage indicators(GEVIs)offers a combination of noninvasiveness,high spatial resolution,and increased measurement throughput.However,its application is limited by the insufficient acquisition rate and time accuracy of the camera.Here we design and apply a stroboscopic illumination scheme to boost the temporal resolution of voltage imaging,while simultaneously eliminating the artifacts caused by nonsynchronized exposure in the rolling-shutter mode.We demonstrate that commonly used GEVIs are compatible with stroboscopic voltage imaging(SVI),and our SVI scheme offers a 5-fold faster acquisition frame rate than that of conventional continuous illumination.The GEVIs tested maintain high sensitivities in the SVI mode,supporting faithful reports of intracellular depolarization waveform and intercellular gap junction-mediated depolarization coupling in human embryonic kidney 293T(HEK 293T)cell populations.SVI allows resolving the action potential(AP)waveform with less distortion and mapping action potential initiation and propagation dynamics in cultured neurons in kilohertz,beyond the restriction from the camera in the field of view.
