摘要
Currently,cellular action potentials are detected using either electrical recordings or exogenous fluorescent probes that sense the calcium concentration or transmembrane voltage.Ca imaging has a low temporal resolution,while voltage indicators are vulnerable to phototoxicity,photobleaching,and heating.Here,we report full-field interferometric imaging of individual action potentials by detecting movement across the entire cell membrane.Using spike-triggered averaging of movies synchronized with electrical recordings,we demonstrate deformations up to 3 nm(0.9 mrad)during the action potential in spiking HEK-293 cells,with a rise time of 4ms.The time course of the optically recorded spikes matches the electrical waveforms.Since the shot noise limit of the camera(~2 mrad/pix)precludes detection of the action potential in a single frame,for all-optical spike detection,images are acquired at 50 kHz,and 50 frames are binned into 1 ms steps to achieve a sensitivity of 0.3 mrad in a single pixel.Using a selfreinforcing sensitivity enhancement algorithm based on iteratively expanding the region of interest for spatial averaging,individual spikes can be detected by matching the previously extracted template of the action potential with the optical recording.This allows all-optical full-field imaging of the propagating action potentials without exogeneous labels or electrodes.
基金
provided by the NIH grant U01 EY025501
by the Stanford Neurosciences Institute(G.G.)。
