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Fast 2-photon stimulation using holographic patterns

Fast 2-photon stimulation using holographic patterns

来源:bioRxiv_logobioRxiv
英文摘要

Two decades after its introduction, optogenetics – a biological technique to control the activity of neurons or other cell types with light – remains a cutting edge and promising tool to study biological processes. Its increasing usage in research varies widely from causally exploring biological mechanisms and neural computations, to neurostimulation and sensory restauration. To stimulate neurons in the brain, a variety of approaches have been developed to generate precise spatiotemporal light patterns. Yet certain constrains still exists in the current optical techniques to activate a neuronal population with both cellular resolution and millisecond precision. Here, we describe an experimental setup allowing to stimulate a few tens of neurons in a plane at sub-millisecond rates using 2-photon activation. A liquid crystal on silicon spatial light modulator (LCoS-SLM) was used to generate spatial patterns in 2 dimensions. The image of the patterns was formed on the plane of a digital micromirror device (DMD) that was used as a fast temporal modulator of each region of interest. Using fluorescent microscopy and patch-clamp recording of neurons in culture expressing the light-gated ion channels, we characterized the temporal and spatial resolution of the microscope. We described the advantages of combining the LCoS-SLM with the DMD to maximize the temporal precision, modulate the illumination amplitude, and reduce background activation. Finally, we showed that this approach can be extended to patterns in 3 dimensions. We concluded that the methodology is well suited to address important questions about the role of temporal information in neuronal coding.

Barral J¨|r¨|mie、Brunstein Maia、Li Wei、Moutoussamy C¨|dric、Lubetzki Jules

Institut Pasteur, Universit¨| Paris Cit¨|, INSERM, Institut de l?ˉAudition||CNRSInstitut Pasteur, Universit¨| Paris Cit¨|, INSERM, Institut de l?ˉAuditionInstitut Pasteur, Universit¨| Paris Cit¨|, INSERM, Institut de l?ˉAuditionInstitut Pasteur, Universit¨| Paris Cit¨|, INSERM, Institut de l?ˉAuditionInstitut Pasteur, Universit¨| Paris Cit¨|, INSERM, Institut de l?ˉAudition

10.1101/2023.01.12.523269

光电子技术生物科学研究方法、生物科学研究技术

Barral J¨|r¨|mie,Brunstein Maia,Li Wei,Moutoussamy C¨|dric,Lubetzki Jules.Fast 2-photon stimulation using holographic patterns[EB/OL].(2025-03-28)[2025-05-17].https://www.biorxiv.org/content/10.1101/2023.01.12.523269.点此复制

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