Neuronal stimulation is essential to understand information processing in brain systems. Spatiotemporal patterns of neuronal activity can be modified by external stimuli. Recent studies have shown that neurons can be stimulated by short-pulse laser processing of the cell membrane. An optical vortex with a helical wavefront possesses an orbital angular momentum (OAM) enables the inward twisting of ablated materials, thereby processing further precisely cells beyond a conventional Gaussian beam. We herein study the mechanisms of neuronal stimulation with a focused nanosecond optical vortex. The focused nanosecond optical vortex on the cell membrane of rat hippocampal neurons induces extracellular Ca2+ influx and neuronal activity elicitation. Morphological changes of the neuronal cell membrane due to nanosecond optical vortex irradiation is also evaluated with fluorescence recovery after photobleaching. After the deposition of a single pulse of nanosecond optical vortex on the cell membrane of neurons, the fluorescence intensity of membrane probe at the focal region significantly decreases, however, it recovers within 5 seconds. Such dynamics suggests that the transient disruption occurs at the cell membrane based on laser ablation and recovers due to lateral diffusion of membrane molecules. The diffusion coefficients of membrane molecules after optical vortex irradiation are larger than those of Gaussian beam irradiation, and the disrupted membrane areas are smaller than the expected ones as the optical vortex focal region. These differences are attributed to the fact that the disruption of cell membrane owing to laser ablation and subsequent membrane diffusion are assisted by OAM transfer effects.
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