A Platform for Single Cell Optogenetics to Study Synaptic Engrams in Vitro

Abstract

Memory relies on the firing of simultaneously activated neurons (engram), whose synapses are strengthened by long-term potentiation mechanisms. Optogenetic tools and a fluorescence probe to map synaptic engrams, were combined with a digital light processor device (DLP), to create in-vitro engrams and study populations of potentiated spines.

Understanding the nervous system: lessons from Frontiers in Neurophotonics

“…On the other hand, for cell culture applications, one can reduce the complexity using a digital light processor (DLP) with an integrated digital micromirror device (DMD) aligned to the EPI-fluorescence port of a spinning disk microscope. Thanks to the DMD, the DLP device can create custom spatial light patterns with a resolution down to ∼3 μm with a 20x objective 42 . This platform can be coupled with optical or electrophysiological monitoring to perform experiments involving the selective excitation/inhibition of a few cells in a neuronal network, followed by the molecular, physiological, and morphological characterization.…”

“…Thanks to the DMD, the DLP device can create custom spatial light patterns with a resolution down to with a 20x objective. 42 This platform can be coupled with optical or electrophysiological monitoring to perform experiments involving the selective excitation/inhibition of a few cells in a neuronal network, followed by the molecular, physiological, and morphological characterization. Reductionistic by design, the platform can be used to investigate basic processes occurring in neuronal cultures at the cellular and synaptic levels, otherwise difficult to address in vivo .…”