Multiplexed aberration measurement for deep tissue imaging in vivo

Wang, Chen; Li, Rui; Milkie, Daniel E.; Sun, Wenzhi; Tan, Zhongchao; Kerlin, Aaron; Chen, Tsai Wen; Kim, Douglas S.; Ji, Na

doi:10.1038/nmeth.3068

Cited by 126 publications

(111 citation statements)

References 20 publications

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“…In IVM, penetration depth is often limited by aberration caused by the sample itself, which may be compensated for by using adaptive optics, for instance using a correction collar (Muriello and Dunn, 2008), a deformable membrane mirror (Caroline Müllenbroich et al, 2014) or segmented pupil illumination (Ji et al, 2012;Wang et al, 2014), which can correct for sample aberrations by increasing signal at depth. Alternatively, image-based metrics can be used to estimate the sample aberrations (Burke et al, 2015;Débarre et al, 2009;Song et al, 2010).…”

Section: Correcting For Sample Aberrationsmentioning

confidence: 99%

Molecular mobility and activity in an intravital imaging setting – implications for cancer progression and targeting

Nobis

Warren

Lucas

et al. 2018

Journal of Cell Science

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Molecular mobility, localisation and spatiotemporal activity are at the core of cell biological processes and deregulation of these dynamic events can underpin disease development and progression. Recent advances in intravital imaging techniques in mice are providing new avenues to study real-time molecular behaviour in intact tissues within a live organism and to gain exciting insights into the intricate regulation of live cell biology at the microscale level. The monitoring of fluorescently labelled proteins and agents can be combined with autofluorescent properties of the microenvironment to provide a comprehensive snapshot of in vivo cell biology. In this Review, we summarise recent intravital microscopy approaches in mice, in processes ranging from normal development and homeostasis to disease progression and treatment in cancer, where we emphasise the utility of intravital imaging to observe dynamic and transient events in vivo. We also highlight the recent integration of advanced subcellular imaging techniques into the intravital imaging pipeline, which can provide in-depth biological information beyond the singlecell level. We conclude with an outlook of ongoing developments in intravital microscopy towards imaging in humans, as well as provide an overview of the challenges the intravital imaging community currently faces and outline potential ways for overcoming these hurdles.

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Section: Correcting For Sample Aberrationsmentioning

confidence: 99%

Molecular mobility and activity in an intravital imaging setting – implications for cancer progression and targeting

Nobis

Warren

Lucas

et al. 2018

Journal of Cell Science

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“…Detectors that combine high frame rate, high sensitivity and large pixel number would be desirable. The SNR of the system, and thus the total number of cells that can be imaged, may also be improved by implementing adaptive optics (Ji et al, 2010(Ji et al, , 2012Wang et al, 2014). This solution would compensate for distortions of the excitation beam introduced along the optical path and within the biological sample, thus improving the efficiency of fluorescence excitation.…”

Section: Scanning and Scanless Imaging Of Neuronal Networkmentioning

confidence: 99%

Optical dissection of brain circuits with patterned illumination through the phase modulation of light

Bovetti

Fellin

2015

Journal of Neuroscience Methods

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Brain function relies on electrical signaling among ensembles of neurons. These signals are encoded in space - neurons are organized in complex three-dimensional networks - and in time-cells generate electrical signals on a millisecond scale. How the spatial and temporal structure of these signals controls higher brain functions is largely unknown. The recent advent of novel molecules that manipulate and monitor electrical activity in genetically identified cells provides, for the first time, the ability to causally test the contribution of specific cell subpopulations in these complex brain phenomena. However, most of the commonly used approaches are limited in their ability to illuminate brain tissue with high spatial and temporal precision. In this review article, we focus on one technique, patterned illumination through the phase modulation of light using liquid crystal spatial light modulators (LC-SLMs), which has the potential to overcome some of the major limitations of current experimental approaches.

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“…However, for most experiments, only a small fraction is used to illuminate a biological sample. For example, when imaging the superficial layers of the murine neocortex with a single scanned excitation spot, an average power between 10 mW (for superficial layer 2/3) and 100 mW (for deep layer 5) is sufficient for functional recordings with the calcium indicator GCaMP6 [7]. As the transmittance of the microscope's optical path can be optimized to >50%, considerable potential exists for less expensive laser sources with a few hundreds of mW output power and with a center wavelength close to the absorption peak of selected fluorophores (Fig.…”

Section: Introductionmentioning

confidence: 99%

Multiphoton in vivo imaging with a femtosecond semiconductor disk laser

Voigt

Emaury

Bethge

et al. 2017

Biomed. Opt. Express

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Abstract:We use an ultrafast diode-pumped semiconductor disk laser (SDL) to demonstrate several applications in multiphoton microscopy. The ultrafast SDL is based on an optically pumped Vertical External Cavity Surface Emitting Laser (VECSEL) passively mode-locked with a semiconductor saturable absorber mirror (SESAM) and generates 170-fs pulses at a center wavelength of 1027 nm with a repetition rate of 1.63 GHz. We demonstrate the suitability of this laser for structural and functional multiphoton in vivo imaging in both Drosophila larvae and mice for a variety of fluorophores (including mKate2, tdTomato, Texas Red, OGB-1, and R-CaMP1.07) and for endogenous second-harmonic generation in muscle cell sarcomeres. We can demonstrate equivalent signal levels compared to a standard 80-MHz Ti:Sapphire laser when we increase the average power by a factor of 4.5 as predicted by theory. In addition, we compare the bleaching properties of both laser systems in fixed Drosophila larvae and find similar bleaching kinetics despite the large difference in pulse repetition rates. Our results highlight the great potential of ultrafast diode-pumped SDLs for creating a cost-efficient and compact alternative light source compared to standard Ti:Sapphire lasers for multiphoton imaging.

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Multiplexed aberration measurement for deep tissue imaging in vivo

Cited by 126 publications

References 20 publications

Molecular mobility and activity in an intravital imaging setting – implications for cancer progression and targeting

Molecular mobility and activity in an intravital imaging setting – implications for cancer progression and targeting

Optical dissection of brain circuits with patterned illumination through the phase modulation of light

Multiphoton in vivo imaging with a femtosecond semiconductor disk laser

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