Optical imaging of metabolic dynamics in animals

Shi, Lingyan; Zheng, Chaogu; Shen, Yihui; Chen, Zhuo; Silveira, Eduardo; Zhang, Luyuan; Wei, Mian; Liu, Chang; Sena-Tomás, Carmen de; Targoff, Kimara L.; Min, Wei

doi:10.1038/s41467-018-05401-3

Cited by 205 publications

(196 citation statements)

References 65 publications

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“…In Fig. C,D, the C–H band can be decomposed to C–H in three cellular components of DNA (2962 cm −1 ), proteins (2928 cm −1 ) and lipids (2874 cm −1 ) (Chen et al , ; Shi et al , ). Accordingly, the C–D shift in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Accordingly, the C-D shift in Fig. 3C,D was associated with the C-D bond-containing DNA (2220 cm À1 ), proteins (2160 cm À1 ) and lipids (2109 cm À1 ) (Chen et al, 2015;Shi et al, 2018). A distinctive pattern of the C-D band was observed using glucose-d 12 compared with D 2 O, with a visibly higher contribution by the lipid component at 2109 cm À1 (Fig.…”

Section: Raman-dip Detected the Microbial Activities In Response To Dmentioning

confidence: 85%

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Raman–deuterium isotope probing to study metabolic activities of single bacterial cells in human intestinal microbiota

Wang

Kong

et al. 2019

Microbial Biotechnology

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Summary Human intestinal microbiota is important to host health and is associated with various diseases. It is a challenge to identify the functions and metabolic activity of microorganisms at the single‐cell level in gut microbial community. In this study, we applied Raman microspectroscopy and deuterium isotope probing (Raman–DIP) to quantitatively measure the metabolic activities of intestinal bacteria from two individuals and analysed lipids and phenylalanine metabolic pathways of functional microorganisms in situ. After anaerobically incubating the human faeces with heavy water (D2O), D2O with specific substrates (glucose, tyrosine, tryptophan and oleic acid) and deuterated glucose, the C–D band in single‐cell Raman spectra appeared in some bacteria in faeces, due to the Raman shift from the C–H band. Such Raman shift was used to indicate the general metabolic activity and the activities in response to the specific substrates. In the two individuals' intestinal microbiota, the structures of the microbial communities were different and the general metabolic activities were 76 ± 1.0% and 30 ± 2.0%. We found that glucose, but not tyrosine, tryptophan and oleic acid, significantly stimulated metabolic activity of the intestinal bacteria. We also demonstrated that the bacteria within microbiota preferably used glucose to synthesize fatty acids in faeces environment, whilst they used glucose to synthesize phenylalanine in laboratory growth environment (e.g. LB medium). Our work provides a useful approach for investigating the metabolic activity in situ and revealing different pathways of human intestinal microbiota at the single‐cell level.

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Section: Resultsmentioning

confidence: 99%

Section: Raman-dip Detected the Microbial Activities In Response To Dmentioning

confidence: 85%

Raman–deuterium isotope probing to study metabolic activities of single bacterial cells in human intestinal microbiota

Wang

Kong

et al. 2019

Microbial Biotechnology

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“…methods developed for volumetric fluorescence imaging alter the tissue composition, and are thus unsuited for comprehensive chemical imaging. Therefore, we identified two Raman-specific criteria, developed a Raman-tailored tissue-clearing strategy, and coupled it with SRS microscopy, including its advanced variations, such as multispectral imaging and deuterium oxide (D 2 O) probing coupled with SRS (DO-SRS) of metabolic probes (33) (Fig. 1A).…”

Section: Significancementioning

confidence: 99%

“…To further study tumor metabolism and angiogenesis, we conducted metabolic imaging using a recently developed method of DO-SRS (33). Going beyond structural imaging, DO-SRS-coupling deuterium oxide probing with SRS microscopycould reveal metabolic dynamics of animals in situ (33). The metabolic incorporation of deuterium from deuterium oxide into macromolecules produces carbon-deuterium bonds that vibrate in the cell-silent spectral window.…”

Section: Metabolic Volumetric Chemical Imaging Of Tumors In the Cell-mentioning

confidence: 99%

Volumetric chemical imaging by clearing-enhanced stimulated Raman scattering microscopy

Wei

Shi

Shen

et al. 2019

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

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115

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Three-dimensional visualization of tissue structures using optical microscopy facilitates the understanding of biological functions. However, optical microscopy is limited in tissue penetration due to severe light scattering. Recently, a series of tissue-clearing techniques have emerged to allow significant depth-extension for fluorescence imaging. Inspired by these advances, we develop a volumetric chemical imaging technique that couples Raman-tailored tissue-clearing with stimulated Raman scattering (SRS) microscopy. Compared with the standard SRS, the clearing-enhanced SRS achieves greater than 10-times depth increase. Based on the extracted spatial distribution of proteins and lipids, our method reveals intricate 3D organizations of tumor spheroids, mouse brain tissues, and tumor xenografts. We further develop volumetric phasor analysis of multispectral SRS images for chemically specific clustering and segmentation in 3D. Moreover, going beyond the conventional label-free paradigm, we demonstrate metabolic volumetric chemical imaging, which allows us to simultaneously map out metabolic activities of protein and lipid synthesis in glioblastoma. Together, these results support volumetric chemical imaging as a valuable tool for elucidating comprehensive 3D structures, compositions, and functions in diverse biological contexts, complementing the prevailing volumetric fluorescence microscopy.

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“…Raman spectroscopy is a powerful analytical tool to detect the frequency shift in the inelastic light scattering due to the molecular vibrational modes in a sample [1,2]. Raman detection is a label-free method [3], thus greatly simplifying the sample preparation and minimally disturbing the sample during detection [4]. It has found a myriad of applications ranging from detection of biohazard contaminants [5], monitoring biological processes [6], disease diagnosis [7], to regenerative medicine [8].…”

Section: Introductionmentioning

confidence: 99%

Raman imaging through multimode sapphire fiber

Deng¹,

Loterie²,

Konstantinou³

et al. 2019

Opt. Express

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We report on a sapphire fiber Raman imaging probe's use for challenging applications where access is severely restricted. Small-dimension Raman probes have been developed previously for various clinical applications because they show great capability for diagnosing disease states in bodily fluids, cells, and tissues. However, applications of these sub-millimeter diameter Raman probes were constrained by two factors: first, it is difficult to incorporate filters and focusing optics at such small scale; second, the weak Raman signal is often obscured by strong background noise from the fiber probe material, especially the most commonly used silica, which has a strong broad background noise in low wavenumbers (<500-1700 cm −1). Here, we demonstrate the thinnest-known imaging Raman probe with a 60 μm diameter Sapphire multimode fiber in which both excitation and signal collection pass through. This probe takes advantage of the low fluorescence and narrow Raman peaks of Sapphire, its inherent high temperature and corrosion resistance, and large numerical aperture (NA). Raman images of Polystyrene beads, carbon nanotubes, and CaSO 4 agglomerations are obtained with a spatial resolution of 1 μm and a field of view of 30 μm. Our imaging results show that single polystyrene bead (~15 µm diameter) can be differentiated from a mixture with CaSO 4 agglomerations, which has a close Raman shift.

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Optical imaging of metabolic dynamics in animals

Cited by 205 publications

References 65 publications

Raman–deuterium isotope probing to study metabolic activities of single bacterial cells in human intestinal microbiota

Raman–deuterium isotope probing to study metabolic activities of single bacterial cells in human intestinal microbiota

Volumetric chemical imaging by clearing-enhanced stimulated Raman scattering microscopy

Raman imaging through multimode sapphire fiber

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