Single cell in vivo optogenetic stimulation by two-photon excitation fluorescence transfer
Lei Tong,
Shanshan Han,
Yao Xue,
Minggang Chen,
Fuyi Chen,
Wei Ke,
Yousheng Shu,
Ning Ding,
Joerg Bewersdorf,
Z. Jimmy Zhou,
Peng Yuan,
Jaime Grutzendler
Affiliations
Lei Tong
Department of Neurology, Yale School of Medicine, New Haven, CT 06511, USA
Shanshan Han
Department of Rehabilitation Medicine, Huashan Hospital, State Key Laboratory of Medical Neurobiology, Institute for Translational Brain Research, MOE Frontiers Center for Brain Science, Fudan University, Shanghai 200032, China
Yao Xue
Department of Ophthalmology and Visual Science, Yale School of Medicine, New Haven, CT 06511, USA
Minggang Chen
Department of Ophthalmology and Visual Science, Yale School of Medicine, New Haven, CT 06511, USA
Fuyi Chen
Department of Neurology, Yale School of Medicine, New Haven, CT 06511, USA
Wei Ke
Department of Neurology, Huashan Hospital, State Key Laboratory of Medical Neurobiology, Institute for Translational Brain Research, MOE Frontiers Center for Brain Science, Fudan University, Shanghai 200032, China
Yousheng Shu
Department of Neurology, Huashan Hospital, State Key Laboratory of Medical Neurobiology, Institute for Translational Brain Research, MOE Frontiers Center for Brain Science, Fudan University, Shanghai 200032, China
Ning Ding
Department of Rehabilitation Medicine, Huashan Hospital, State Key Laboratory of Medical Neurobiology, Institute for Translational Brain Research, MOE Frontiers Center for Brain Science, Fudan University, Shanghai 200032, China
Joerg Bewersdorf
Department of Cell Biology, Yale School of Medicine, New Haven, CT 06511, USA; Department of Biomedical Engineering, Yale University, New Haven, CT 06511, USA
Z. Jimmy Zhou
Department of Ophthalmology and Visual Science, Yale School of Medicine, New Haven, CT 06511, USA; Department of Neuroscience, Yale School of Medicine, New Haven, CT 06511, USA; Department of Cellular and Molecular Physiology, Yale University, New Haven, CT 06511, USA
Peng Yuan
Department of Neurology, Yale School of Medicine, New Haven, CT 06511, USA; Department of Rehabilitation Medicine, Huashan Hospital, State Key Laboratory of Medical Neurobiology, Institute for Translational Brain Research, MOE Frontiers Center for Brain Science, Fudan University, Shanghai 200032, China; Corresponding author
Jaime Grutzendler
Department of Neurology, Yale School of Medicine, New Haven, CT 06511, USA; Department of Neuroscience, Yale School of Medicine, New Haven, CT 06511, USA; Corresponding author
Summary: Optogenetic manipulation with single-cell resolution can be achieved by two-photon excitation. However, this frequently requires relatively high laser powers. Here, we developed a novel strategy that can improve the efficiency of current two-photon stimulation technologies by positioning fluorescent proteins or small fluorescent molecules with high two-photon cross-sections in the vicinity of opsins. This generates a highly localized source of endogenous single-photon illumination that can be tailored to match the optimal opsin absorbance. Through neuronal and vascular stimulation in the live mouse brain, we demonstrate the utility of this technique to achieve efficient opsin stimulation, without loss of cellular resolution. We also provide a theoretical framework for understanding the potential advantages and constrains of this methodology, with directions for future improvements. Altogether, this fluorescence transfer illumination method opens new possibilities for experiments difficult to implement in the live brain such as all-optical neural interrogation and control of regional cerebral blood flow.
