In this presentation, faculty will provide information to optimize, validate, and troubleshoot optogenetics experiments in order to avoid and address potential confounds. Specifically, Julia Lemos and Shana Augustin will: - Describe how to achieve optimal opsin expression. - Describe how to design control experiments to optimize opsin expression while limiting confounds. - Describe anatomical and functional validation of opsin expression. After watching this presentation, you should be able to identify and understand how to address common potential pitfalls in optogenetics experiments.

In this presentation, faculty cover some of the key technical issues that arise when designing and interpreting optogenetic experiments. Specifically, Chris Chen and Scott Owen will: - Describe how to select optimal tools and stimulation parameters for an optogenetics experiment. - Identify potential confounds related to stimulus parameters. - Describe key methods to validate optogenetic tools. After watching this presentation, you should be able to better understand how to select optimal tools and stimulation parameters for an optogenetics experiment.

In this presentation, Stephanie Cragg and Yevgenia Kozorovitskiy will cover the basics of using optogenetics in acute brain slices, also called ex vivo. Specifically, they will: - Identify key concepts in the design and interpretation of ex vivo optogenetic experiments. - Describe critical parts and equipment for ex vivo optogenetics experiments. After watching this presentation, you should be able to understand the basic setup and key concepts for ex vivo optogenetics experiments.

In this presentation, Mario Penzo will describe the various strategies commonly employed for the delivery of bacterial opsins to the nervous system. Specifically, Penzo will: - Describe different opsin delivery methods, such as virus injection, electroporation, and use of transgenic mice. - Identify, for a particular delivery strategy (adeno-associated virus vectors), the importance of the various serotypes available. After watching this presentation, you should be able to understand the pros and cons of the different opsin delivery methods.

In this presentation, faculty cover some of the major caveats to optogenetics experiments, at the conceptual and technical level. Specifically: - Stephan Lammel discusses the importance of careful methodological considerations for interpreting the results of optogenetic experiments. - Karel Svoboda highlights some of the many potential unexpected effects of optogenetics perturbation in the context of neural circuit dynamics and behavior. - David Kupferschmidt shares how diverse forms of synaptic plasticity and complex neurochemistry can complicate our ability to infer direct “causation” from optogenetics experiments. After watching this presentation, you should be able to: - Understand and control for potential confounds of optogenetics manipulations that involve non-cell-type-specific expression of opsins, toxicity, tissue heating, and scattered light delivery. - Understand how to turn potential unexpected effects of optogenetics manipulation to your advantage using neurophysiological characterization of the system response optogenetics perturbation. - Understand and minimize the potential confounds of synaptic plasticity and complex neurochemistry in the interpretation of optogenetics experiments.