Three-dimensional live multi-label light-sheet imaging with synchronous excitation-multiplexed structured illumination

Xu, Dongli; Zhou, Weibin; Peng, Leilei

doi:10.1364/oe.25.031159

Cited by 7 publications

(3 citation statements)

References 30 publications

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“…17 Different LSFM configurations have been proposed to perform multicolor 3-D imaging. [18][19][20][21] For instance, Krieger et al performed dual color fluorescence imaging using a single camera and two separate color channels, while Jahr et al used a diffractive unit to spectrally split the images onto a camera in order to obtain hyperspectral images. Additionally, Mahou et al implemented a two-photon multicolor light-sheet microscope using a femtosecond laser and an optical parametric oscillator to obtain multicolor two-photon excitation using a single camera.…”

Section: Introductionmentioning

confidence: 99%

Multicolor fluorescence microscopy using static light sheets and a single-channel detection

et al. 2019

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We present a multicolor fluorescence microscope system, under a selective plane illumination microscopy (SPIM) configuration, using three continuous wave-lasers and a single-channel-detection camera. The laser intensities are modulated with three time-delayed pulse trains that operate synchronously at one third of the camera frame rate, allowing a sequential excitation and an image acquisition of up to three different biomarkers. The feasibility of this imaging acquisition mode is demonstrated by acquiring single-plane multicolor images of living hyphae of Neurospora crassa. This allows visualizing simultaneously the localization and dynamics of different cellular components involved in apical growth in living hyphae. The configuration presented represents a noncommercial, cost-effective alternative microscopy system for the rapid and simultaneous acquisition of multifluorescent images and can be potentially useful for three-dimensional imaging of large biological samples.

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Section: Introductionmentioning

confidence: 99%

Multicolor fluorescence microscopy using static light sheets and a single-channel detection

et al. 2019

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“…Line detection techniques have also found wide applications in chemistry [6] and food inspection [10,11]. Recently, the light-sheet microscopy (LSM) technique, which illuminates a transparent sample with a thin sheet of laser light and images the sample orthogonally, facilitates rapid volumetric imaging [12][13][14][15][16][17]. The thin sheet of laser light can be static, generated by a cylindrical lens [12], or virtual, generated by a fast-scanning galvo-mirror [13,14].…”

Section: Introductionmentioning

confidence: 99%

Exploratory Study on Light-Sheet Based Three-Dimensional Surface Topography

Cai¹,

Chen²,

Zhou³

et al. 2018

PIER

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Light-sheet microscopy has attracted considerable attention because it is a fluorescence imaging technique with rapid optical sectioning capability for transparent samples. In this study, we report a new application based on light-sheet microscopy for exploratory investigation of threedimensional surface topography of opaque objects. Instead of using inelastic scattering fluorescent signals, our method utilizes the elastic scattering of light from the surface of opaque samples, which are illuminated by a light sheet generated by a cylindrical lens. Through a simple structural modification by removing the fluorescent filter, the orthogonal imaging module can capture the elastically-scattered image. As the opaque object is scanned by a motorized stage, the light-sheet microscope acquires a series of sectional images, which can be stitched into a three-dimensional surface topography image. This method also offers the opportunity to visualize a 3D fingerprint at micron-level resolution. Therefore, this technique may be used in industry and the biomedical field for the measurement of surface microstructure. To our best knowledge, this is the first time a light-sheet microscopy is utilized to perform surface topography measurement.

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“…The penetration depth can be further increased using Bessel (19,20) or Airy beams (21). Structured illumination has been used to modulate the excitation of Gaussian beam (22), Bessel beam (23)(24)(25), or lattice pattern (2,14), providing a different approach to remove scattered fluorescence light, reject out-of-focus background, and enable thinner optical sectioning. Superresolution can be realized using structured illumination microscopy (SIM) (2,(12)(13)(14) by extracting high-frequency details embedded in low-resolution moiré fringes imaged under an illumination pattern at several shifted phases (26).…”

mentioning

confidence: 99%

In vivo NIR-II structured-illumination light-sheet microscopy

Wang

Zhong

et al. 2021

Proc. Natl. Acad. Sci. U.S.A.

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Noninvasive optical imaging with deep tissue penetration depth and high spatiotemporal resolution is important to longitudinally studying the biology at the single-cell level in live mammals, but has been challenging due to light scattering. Here, we developed near-infrared II (NIR-II) (1,000 to 1,700 nm) structured-illumination light-sheet microscopy (NIR-II SIM) with ultralong excitation and emission wavelengths up to ∼1,540 and ∼1,700 nm, respectively, suppressing light scattering to afford large volumetric three-dimensional (3D) imaging of tissues with deep-axial penetration depths. Integrating structured illumination into NIR-II light-sheet microscopy further diminished background and improved spatial resolution by approximately twofold. In vivo oblique NIR-II SIM was performed noninvasively for 3D volumetric multiplexed molecular imaging of the CT26 tumor microenvironment in mice, longitudinally mapping out CD4, CD8, and OX40 at the single-cell level in response to immunotherapy by cytosine-phosphate-guanine (CpG), a Toll-like receptor 9 (TLR-9) agonist combined with OX40 antibody treatment. NIR-II SIM affords an additional tool for noninvasive volumetric molecular imaging of immune cells in live mammals.

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Three-dimensional live multi-label light-sheet imaging with synchronous excitation-multiplexed structured illumination

Cited by 7 publications

References 30 publications

Multicolor fluorescence microscopy using static light sheets and a single-channel detection

Multicolor fluorescence microscopy using static light sheets and a single-channel detection

Exploratory Study on Light-Sheet Based Three-Dimensional Surface Topography

In vivo NIR-II structured-illumination light-sheet microscopy

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