Optogenetics PDF: A Roadmap
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Optogenetics—as the name suggests—coalesces multiple disciplines in Biomedicine, Optics, and Biotechnology, and now, almost 12 years after the publication of the first seminal paper, it may be safe to state that optogenetics truly revolutionized neuroscience. For the first time, a causal interrogation of neuronal circuitry with millisecond precision, cell-type specificity, and minimal invasiveness became a reality. As a result, optogenetic techniques have pervaded almost all disciplines and fields of neuroscience over the years, from applications in brain slices to in vivo application in rodents , zebrafish, C. elegans , and nonhuman primates , and with clinical applications now just on the horizon. What is more, optogenetics allowed for the combination with already well-established readouts, such as single-cell electrophysiology , electric population recordings, functional magnetic resonance imaging, and, more recently, optical methods such as 2-photon calcium imaging. The research questions involving optogenetics are equally diverse, including the modulation of stem cell differentiation in mouse brain, of C. elegans behavior, and the causal dissection of sensory processing in nonhuman primates. While many scholarly articles and books exist on each of these methods and animal models , from viral vectors to electrophysiology and behavior, there are unique requirements for each step of the optogenetic approach. In fMRI, for instance, imaging coils need a lead-through for an optic fiber. Likewise, viral vectors need to accommodate the sequences of promoter , opsin , and fluorophore. Some research questions may not require any sophisticated cellular targeting tools, but rather the implementation of a complicated readout interfering with optical interrogation, while others may center on a novel promoter in combination with fairly standard opsins and analytical tools. And maybe most importantly, a project involving optogenetics has to entail tailored control experiments addressing both a potential interaction of light with the readout method and the potential effect of light itself on neuronal physiology. Still, even after more than a decade, a successful implementation of optogenetics requires expertise in multiple fields, and each step of the way poses individual challenges and takes considerable time and efforts. This book should guide both the optogenetics newbie and the expert through all the steps required for the implementation of optogenetics in neuroscience. It should empower the reader to identify the critical aspects of each methodological step and to decide on the necessary level of complexity to address the respective research question.
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