Miniaturized microscope with flexible light source input for neuronal imaging and manipulation in freely behaving animals

Simultaneous imaging and manipulation of a genetically defined neuronal population can provide a causal link between its activity and function. Here, we designed a miniaturized microscope (or ‘miniscope’) that allows fluorescence imaging and optogenetic manipulation at the cellular level in freely behaving animals. This miniscope has an integrated optical connector that accepts any combination of external light sources, allowing flexibility in the choice of sensors and manipulators. Moreover, due to its simple structure and use of open source software, the miniscope is easy to build and modify. Using this miniscope, we demonstrate the optogenetic silencing of hippocampal CA1 neurons using two laser light sources—one stimulating a calcium sensor (i.e., jGCaAMP7c) and the other serving as an optogenetic silencer (i.e., Jaws). This new miniscope can contribute to efforts to determine causal relationships between neuronal network dynamics and animal behavior. •We developed a miniaturized microscope capable of simultaneously imaging and manipulating.•It requires minimum alterations from an open-sourced miniaturized microscope (UCLA-miniscope).•It can be used in the freely-moving condition.•It accepts any combinations of external light sources.•It allows flexibility in the combination of light-operated sensors.