Multicolor Stimulated Raman Scattering Microscopy With Fast Wavelength-Tunable Yb Fiber Laser

Ozeki, Yasuyuki; Asai, Takuya; Shou, Jingwen; Yoshimi, Hironobu

doi:10.1109/jstqe.2018.2830816

Cited by 90 publications

(64 citation statements)

References 64 publications

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“…Figure 3c shows the RIACS images of Euglena gracilis cells cultured in NaH 13 CO 3 medium and those cultured in NaH 12 CO 3 medium, which exhibit granular morphology of 13 C-paramylon (red) and 12 C-paramylon (green), respectively. Figure 3d shows that hiPSCs exhibit a stronger carbohydrate signal after treatment, which is consistent with SRS images ( Supplementary Figure 6b) taken with our SRS microscope 65 .…”

Section: Methodssupporting

confidence: 85%

Raman image-activated cell sorting

et al. 2020

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The advent of image-activated cell sorting and imaging-based cell picking has advanced our knowledge and exploitation of biological systems in the last decade. Unfortunately, they generally rely on fluorescent labeling for cellular phenotyping, an indirect measure of the molecular landscape in the cell, which has critical limitations. Here we demonstrate Raman image-activated cell sorting by directly probing chemically specific intracellular molecular vibrations via ultrafast multicolor stimulated Raman scattering (SRS) microscopy for cellular phenotyping. Specifically, the technology enables real-time SRS-image-based sorting of single live cells with a throughput of up to~100 events per second without the need for fluorescent labeling. To show the broad utility of the technology, we show its applicability to diverse cell types and sizes. The technology is highly versatile and holds promise for numerous applications that are previously difficult or undesirable with fluorescence-based technologies.

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Section: Methodssupporting

confidence: 85%

Raman image-activated cell sorting

et al. 2020

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“…2 A , SRS images of the beads at 4 Raman wavenumbers indicate their characteristic signatures at the different wavenumbers. These 4-color Raman spectra show a good agreement with those obtained by hyperspectral SRS microscopy (31, 32) (Fig. 2 B ).…”

Section: Resultssupporting

confidence: 80%

“…The time-delayed spectral components are partly amplified by bidirectional ytterbium-doped fiber amplifiers (Bi-YDFAs) and recombined to generate a 4-color pulse train. Compared with our previous galvanometric scanner-based wavelength tuning technique (31, 32), the present nonmechanical wavelength-switchable laser is 4 orders of magnitude faster, enabling multicolor SRS imaging of flowing cells.…”

Section: Resultsmentioning

confidence: 94%

Label-free chemical imaging flow cytometry by high-speed multicolor stimulated Raman scattering

Suzuki

Kobayashi

Wakisaka

et al. 2019

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

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166

132

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Combining the strength of flow cytometry with fluorescence imaging and digital image analysis, imaging flow cytometry is a powerful tool in diverse fields including cancer biology, immunology, drug discovery, microbiology, and metabolic engineering. It enables measurements and statistical analyses of chemical, structural, and morphological phenotypes of numerous living cells to provide systematic insights into biological processes. However, its utility is constrained by its requirement of fluorescent labeling for phenotyping. Here we present label-free chemical imaging flow cytometry to overcome the issue. It builds on a pulse pair-resolved wavelength-switchable Stokes laser for the fastest-to-date multicolor stimulated Raman scattering (SRS) microscopy of fast-flowing cells on a 3D acoustic focusing microfluidic chip, enabling an unprecedented throughput of up to ∼140 cells/s. To show its broad utility, we use the SRS imaging flow cytometry with the aid of deep learning to study the metabolic heterogeneity of microalgal cells and perform marker-free cancer detection in blood.

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“…Advances in cryo-EM [158] or other novel crystallography methods [159] pioneered by many research labs in Japan could allow for native structural elucidation of reaction dynamics within protocells [160,161], ribonucleopeptides [162,163], or primitive compartments [164]. Finally, developments in stimulated Raman scattering microscopy [165,166] and superresolution microscopy [167,168] applied towards protocellular studies could further provide spatial insights of reactions within primitive protocells with high resolution.…”

Section: The Future Of Ool Research: Incorporating Novel Biophysics Tmentioning

confidence: 99%

Recent Advances in Origins of Life Research by Biophysicists in Japan

Jia

Kuruma

2019

Challenges

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Biophysics research tends to focus on utilizing multidisciplinary technologies and interdisciplinary collaborations to study biological phenomena through the lens of chemistry and physics. Although most current biophysics work is focused on studying extant biology, the fact remains that modern biological systems at some point were descended from a universal common ancestor. At the core of modern biology is the important question of how the earliest life on (or off) Earth emerged. Recent technological and methodological advances developed by biophysicists in Japan have allowed researchers to gain a new suite of knowledge related to the origins of life (OoL). Using these reports as inspiration, here, we highlight some of the significant OoL advances contributed by members of the biophysical research field in Japan with respect to the synthesis and assembly of biological (or pre-biological) components on early Earth, the co-assembly of primitive compartments with biopolymer systems, and the evolution of early genetic systems. We hope to provide inspiration to other biophysicists to not only use the always-advancing suite of available multidisciplinary technologies to continue their own line of work, but to also consider how their work or techniques can contribute to the ever-growing field of OoL research.

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Multicolor Stimulated Raman Scattering Microscopy With Fast Wavelength-Tunable Yb Fiber Laser

Cited by 90 publications

References 64 publications

Raman image-activated cell sorting

Raman image-activated cell sorting

Label-free chemical imaging flow cytometry by high-speed multicolor stimulated Raman scattering

Recent Advances in Origins of Life Research by Biophysicists in Japan

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