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Neuron. 2013 Nov 20;80(4):900-13. doi: 10.1016/j.neuron.2013.07.052. Epub 2013 Oct 17.

Chronic cellular imaging of entire cortical columns in awake mice using microprisms.

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Division of Endocrinology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, CLS750, 330 Brookline Avenue, Boston, MA 02215, USA; Department of Neurobiology, Harvard Medical School, Goldenson 243, 220 Longwood Avenue, Boston, MA 02115 USA. Electronic address:


Two-photon imaging of cortical neurons in vivo has provided unique insights into the structure, function, and plasticity of cortical networks, but this method does not currently allow simultaneous imaging of neurons in the superficial and deepest cortical layers. Here, we describe a simple modification that enables simultaneous, long-term imaging of all cortical layers. Using a chronically implanted glass microprism in barrel cortex, we could image the same fluorescently labeled deep-layer pyramidal neurons across their entire somatodendritic axis for several months. We could also image visually evoked and endogenous calcium activity in hundreds of cell bodies or long-range axon terminals, across all six layers in visual cortex of awake mice. Electrophysiology and calcium imaging of evoked and endogenous activity near the prism face were consistent across days and comparable with previous observations. These experiments extend the reach of in vivo two-photon imaging to chronic, simultaneous monitoring of entire cortical columns.

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