Johannes Vierock,#1 Silvia Rodriguez-Rozada,#2 Alexander Dieter,2 Florian Pieper,3 Ruth Sims,4 Federico Tenedini,5 Amelie C. F. Bergs,6 Imane Bendifallah,4 Fangmin Zhou,5 Nadja Zeitzschel,6 Joachim Ahlbeck,3 Sandra Augustin,1 Kathrin Sauter,2,5 Eirini Papagiakoumou,4 Alexander Gottschalk,6 Peter Soba,5,7 Valentina Emiliani,4 Andreas K. Engel,3 Peter Hegemann,1 and J. Simon Wiegertcorresponding author2


1Institute for Biology, Experimental Biophysics, Humboldt University Berlin, Berlin, Germany
2Research Group Synaptic Wiring and Information Processing, Center for Molecular Neurobiology Hamburg, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
3Department of Neurophysiology and Pathophysiology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
4Wavefront-Engineering Microscopy Group, Photonics Department, Institut de la Vision, Sorbonne Université, INSERM, CNRS, Institut de la Vision, Paris, France
5Research Group Neuronal Patterning and Connectivity, Center for Molecular Neurobiology Hamburg, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
6Buchmann Institute for Molecular Life Sciences and Institute of Biophysical Chemistry, Goethe University, Frankfurt, Germany
7LIMES Institute, University of Bonn, Bonn, Germany
J. Simon Wiegert, Email: [email protected]
corresponding authorCorresponding author.
#Contributed equally.


Neuroscience, Optogenetics


Optogenetic manipulation of neuronal activity through excitatory and inhibitory opsins has become an indispensable experimental strategy in neuroscience research. For many applications bidirectional control of neuronal activity allowing both excitation and inhibition of the same neurons in a single experiment is desired. This requires low spectral overlap between the excitatory and inhibitory opsin, matched photocurrent amplitudes and a fixed expression ratio. Moreover, independent activation of two distinct neuronal populations with different optogenetic actuators is still challenging due to blue-light sensitivity of all opsins. Here we report BiPOLES, an optogenetic tool for potent neuronal excitation and inhibition with light of two different wavelengths. BiPOLES enables sensitive, reliable dual-color neuronal spiking and silencing with single- or two-photon excitation, optical tuning of the membrane voltage, and independent optogenetic control of two neuronal populations using a second, blue-light sensitive opsin. The utility of BiPOLES is demonstrated in worms, flies, mice and ferrets.



Monochromatic light (± 7 nm) was provided by a Polychrome V monochromator (TILL Photonics, Planegg, Germany) or by a pE-4000 CoolLED system (CoolLED, Andover, UK) for light titration experiments.

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The pE-4000 Universal Illumination System offers 16 selectable wavelengths from 365 - 770 nm, making it a highly flexible illuminator covering a wide variety of fluorophores

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Nature Communications

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