2026/05/12 by Mary L. Phillips, Nicolai T. Urban, Taddeo Salemi +2 · 1 voice
Biochemistry, Genetics and Molecular Biology · Neuroscience · #Advanced Fluorescence Microscopy Techniques #Cell Image Analysis Techniques #Photoreceptor and optogenetics research
paper · doi:10.7554/elife.110277.3
openalex created_date 2023/12/15 · openalex publication_date 2026/05/12 · openalex updated_date 2026/08/01
Abstract Head-mounted miniscopes have enabled functional fluorescence imaging in freely moving animals. However, current technology is limited to recording at most two spectrally distinct fluorophores, severely restricting the number of identifiable cell types. Here we introduce multiplexed neuronal imaging (Neuroplex), a pipeline combining miniscope Ca 2+ recordings with in vivo multiplexed confocal spectral imaging to distinguish nine projection-defined neuronal subtypes through the same GRIN lens. By co-registering defined neurons with fluorophore-specific spectral fingerprints via linear unmixing, we link projection-defined identities to behaviorally relevant neuronal activity. This approach overcomes spectral constraints of miniscopes, enabling circuit-level dissection of behavior in single animals.