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Summary: An extensive literature describes how pupil size reflects neuromodulatory activity, including the noradrenergic system. Here, we present a protocol for the simultaneous recording of optogenetically identified locus coeruleus (LC) units and pupil diameter in mice under different conditions. We describe steps for building an optrode, performing surgery to implant the optrode and headpost, searching for opto-tagged LC units, and performing dual LC-pupil recording. We then detail procedures for data processing and analysis.For complete details on the use and execution of this protocol, please refer to Megemont et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.