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This lecture explores light- and calcium-gated tools to probe neuronal activity, focusing on optogenetics. It discusses the principles behind optogenetics, the development of light-responsive proteins, and their applications in studying brain information processing and potential clinical uses. The lecture also covers neurotransmission basics, genetically-encoded tools for neuronal activity probing, and the design of fusion proteins for optimal light-controlled cleavage. Furthermore, it delves into the use of directed evolution to engineer proteins for specific functions and the application of flow cytometry for single-cell analysis. The learning goals include understanding neuronal communication through electrical and chemical signals, designing methods to probe neuronal activity, and setting up directed evolution experiments.