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Two-photon photostimulation and imaging of neural circuits


We introduce an optical method to stimulate individual neurons in brain slices in any arbitrary spatiotemporal pattern, using two-photon uncaging of MNI-glutamate with beam multiplexing. This method has single-cell and three-dimensional precision. By sequentially stimulating up to a thousand potential presynaptic neurons, we generated detailed functional maps of inputs to a cell. We combined this approach with two-photon calcium imaging in an all-optical method to image and manipulate circuit activity.

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Figure 1: Targeted imaging of neuronal populations.
Figure 2: Two-photon photostimulation with beam multiplexing.
Figure 3: Mapping inputs with single cell resolution.
Figure 4: Mapping inputs in three dimensions.
Figure 5: Reproducibility of input maps.
Figure 6: All-optical stimulation and imaging of network activity.

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We thank E. Callaway for advice, A. Packer for online analysis, B. Nemet for help with optics, G. Ellis-Davies for initial samples of MNI-glutamate, D. Aronov for the convex hull algorithm, Y. Duanmu for histology, and L. Abbott, E. Schaffer and members of the laboratory for comments. Supported by the US National Eye Institute, the National Institute of Neurological Disorders and Stroke, the New York State Foundation for Science Technology and Innovation (NYSTAR) program and the Kavli Institute for Brain Studies. K.E.P. is a Patterson Trust postdoctoral fellow. We dedicate this study to the memory of Larry Katz, who pioneered and stimulated these experiments.

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Correspondence to Volodymyr Nikolenko.

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Supplementary figures 1–5, Supplementary Note, Supplementary Methods (PDF 1033 kb)

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Nikolenko, V., Poskanzer, K. & Yuste, R. Two-photon photostimulation and imaging of neural circuits. Nat Methods 4, 943–950 (2007).

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