Abstract
The generation and application of human stem-cell-derived functional neural circuits promises novel insights into neurodegenerative diseases. These networks are often studied using stem-cell derived random neural networks in vitro, with electrical stimulation and recording using multielectrode arrays. However, the impulse response function of networks is best obtained with spatiotemporally well-defined stimuli, which electrical stimulation does not provide. Optogenetics allows for the functional control of genetically altered cells with light stimuli at high spatiotemporal resolution. Current optogenetic investigations of neural networks are often conducted using full field illumination, potentially masking important functional information. This can be avoided using holographically shaped illumination. In this article, we present a digital holographic illumination setup with a spatial resolution of about 8 µm, which suffices for the stimulation of single neurons, and offers a temporal resolution of less than 0.6 ms. With this setup, we present preliminary single-cell stimulation recording of stem-cell derived induced human neurons in a random neural network. This will offer the opportunity for further studies on connectivity in such networks.
Highlights
Based on powerful imaging techniques, such as functional magnetic resonance imaging, brain research has made great progress in recent years [1,2,3,4,5,6,7]
Since insights that are gained in animal models cannot fully be transferred to humans [8], it is of great interest to perform studies directly in human neural networks
The aim of our work presented in this paper is to provide a stimulation setup for the investigation of neural networks derived from human induced pluripotent stem cells that are tagged with optogenetic tools [12]
Summary
Based on powerful imaging techniques, such as functional magnetic resonance imaging, brain research has made great progress in recent years [1,2,3,4,5,6,7]. The aim of our work presented in this paper is to provide a stimulation setup for the investigation of neural networks derived from human induced pluripotent stem cells that are tagged with optogenetic tools [12]. E.g., signal transport in neural networks, can be more obtained by exciting neurons directly Both patch-clamp and extracellular electrodes can provide electrical stimulation and consecutive recording. The first requirement can be achieved by an illumination setup with steering mirror galvanometers or acousto-optic deflectors These have been used for activating cardiomyocytes [20] or for two-photon excitation and imaging of neural networks [21,22,23]. Simultaneous stimulation is possible by actively shaping the illumination beam using spatial light modulators (SLM). Are derived from human induced pluripotent stem cells, using this setup as a pathway to a deeper understanding of such neural networks
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