2020
DOI: 10.1101/2020.04.27.063388
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Volumetric two-photon fluorescence imaging of live neurons using a multimode optical fiber

Abstract: 11An ability to optically visualize neurons and their subcellular components across brain regions is 12 integral to our understanding of neuronal function 1 . Multimode optical fibers (MMFs), combined 13 with wavefront control methods, have achieved minimally-invasive in vivo imaging of neurons in 14 deep-brain regions with diffraction-limited spatial resolution 2-5 . To date MMF systems have been 15 based on linear fluorescence and are therefore only able to perform two-dimensional imaging due to 16 an absenc… Show more

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Cited by 3 publications
(5 citation statements)
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“…Second, the ability to capture the morphology of subcellular objects, such as dendritic spines on neurons, is an important capability of MMF imaging systems, which will primarily be enabled through a substantial increase in NA. Third, advances in two-photon excited fluorescence microscopy through MMF will benefit from the use of high NA because of the nonlinear dependence between signal generation and NA [21,29]. In fact, even at equivalent NA the non-linearity of the two-photon process will result in increased spatial variations compared to one-photon fluorescence because the excitation volume is effectively the square of the illumination volume.…”
Section: Discussionmentioning
confidence: 99%
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“…Second, the ability to capture the morphology of subcellular objects, such as dendritic spines on neurons, is an important capability of MMF imaging systems, which will primarily be enabled through a substantial increase in NA. Third, advances in two-photon excited fluorescence microscopy through MMF will benefit from the use of high NA because of the nonlinear dependence between signal generation and NA [21,29]. In fact, even at equivalent NA the non-linearity of the two-photon process will result in increased spatial variations compared to one-photon fluorescence because the excitation volume is effectively the square of the illumination volume.…”
Section: Discussionmentioning
confidence: 99%
“…Organotypic hippocampal slices (350 µm) were prepared from male Wistar rats (P7-P8; Harlan UK) as previously described [ 21 , 25 ]. After dissection, slices were cultured on Millicell CM membranes and maintained in culture media at C for 7-14 days prior to use.…”
Section: Methodsmentioning
confidence: 99%
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“…Unfortunately, combining ultrashort pulses with MMF imaging is non-trivial, as the modal interference and modal dispersion in a MMF results in a complex spatiotemporal output field [12,13]. Recently, several nonlinear optical imaging techniques through a single MMF probe have been demonstrated including two-photon excitation microscopy [14,15], 3D microfabrication based on two-photon polymerization [16], and coherent anti-Stokes Raman scattering (CARS) microscopy [17]. All these methods of nonlinear imaging require spatial-domain wavefront shaping and consequently control of many spatial modes on the MMF input.…”
Section: Introductionmentioning
confidence: 99%
“…Interestingly, the transverse wave-vector conservation naturally holds in step-index (SI) MMF [16]. Furthermore, broadband light propagation through a MMF has recently been observed resulting in an axially extended focus allowing efficient volumetric imaging [30,31]. In this letter we experimentally characterize the χ-axial ME, so far unexplored, at the distal facets of SI-MMFs, and theoretically model it based on the study of the correlation function.…”
mentioning
confidence: 99%