Apr 28, 2020

Two-photon fluorescence imaging of live neurons using a multimode optical fiber

BioRxiv : the Preprint Server for Biology
R. TurcotteNigel J. Emptage

Abstract

An ability to optically visualize neurons and their subcellular components across brain regions is integral to our understanding of neuronal function. Multimode optical fibers (MMFs), combined with wavefront control methods, have achieved minimally-invasive in vivo imaging of neurons in deep-brain regions with diffraction-limited spatial resolution. To date MMF systems have been based on linear fluorescence and are therefore only able to perform two-dimensional imaging due to an absence of confocal filtering. Here we report a method to achieve optical sectioning in live brain tissue by taking advantage of the nonlinearity of two-photon excited fluorescence (TPEF). In addition, we provide a new approach for the determination of transmission matrix for TPEF, a necessary step for the light shaping requirement of MMF imaging. Central to these advances is the use of a laser source able to generate both continuous wave (CW) light and femtosecond pulses. This configuration allows the system to be compact with minimal distal optics, enhancing suitability for neuroscience applications. With our system we demonstrate TPEF imaging of neurons and their dendrites in live brain slices through a single 60 m-core MMF with a numerical aperture ...Continue Reading

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