In Situ and In Vivo Molecular Analysis by Coherent Raman Scattering Microscopy

Liao, Chang; Cheng, Ji‐Xin

doi:10.1146/annurev-anchem-071015-041627

Cited by 44 publications

(24 citation statements)

References 148 publications

(205 reference statements)

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“…In comparison with spectral imaging, by which multiple wavelengthdependent images are obtained, spectroscopic imaging aims to achieve the vibrational-modedependent deconvolution of images with spectral data sets. The use of spectroscopic imaging has become prominent for coherent or nonlinear Raman imaging methods [1][2][3][4][5][6][7][8][9], among which coherent anti-Stokes Raman scattering (CARS) and stimulated Raman scattering (SRS) are two methods that are often used. Based on the CARS and SRS imaging techniques, several kinds of spectroscopic imaging have been reported, such as the multiplex method [10][11][12][13][14][15][16], laser-wavelength-scanning method [17][18][19], spectral focusing method [20], FT-CARS [21][22][23], and dual-comb method [24].…”

Section: Introductionmentioning

confidence: 99%

Ultra-multiplex CARS spectroscopic imaging with 1-millisecond pixel dwell time

Kano¹,

Maruyama²,

Kano³

et al. 2019

OSA Continuum

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We performed ultra-multiplex coherent anti-Stokes Raman scattering (CARS) spectroscopic imaging by using a CCD camera with a fast readout time (<1 ms). The ultra-multiplex CARS signal of a polystyrene bead was detected in the range 600-3600 cm −1 with resolution <10 cm −1. The pixel dwell time was approximately 1 ms, which was limited by the readout time of the CCD camera rather than the exposure time. CARS images of 161 × 161 pixels were obtained of the polymer beads with a total data-acquisition time of approximately 28 s even with the use of a cost-effective microchip laser source. Label-free and ultra-multiplex (18 colors) imaging of living cells was also performed with an effective exposure time of 1.8 ms based on the molecular fingerprint as well as the C-H and O-H stretching vibrational modes using a master oscillator fiber amplifier laser source.

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Section: Introductionmentioning

confidence: 99%

Ultra-multiplex CARS spectroscopic imaging with 1-millisecond pixel dwell time

Kano¹,

Maruyama²,

Kano³

et al. 2019

OSA Continuum

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“…Vibrational imaging by SRS is a rapidly growing field. [20][21][22][23][24] It is becoming the most powerful vibrational microscopy for biology, owing to its superb sensitivity, imaging speed, three-dimensional optical sectioning, Raman spectral fidelity, a strict linear concentration dependence, straightforward image interpretation, and quantification. [20][21][22][23][24][25][26][27][28] Recently, a new bioorthogonal imaging strategy is emerging, by introducing vibrational tags such as deuterium or alkyne labels to small biomolecules.…”

Section: Introductionmentioning

confidence: 99%

“…[20][21][22][23][24] It is becoming the most powerful vibrational microscopy for biology, owing to its superb sensitivity, imaging speed, three-dimensional optical sectioning, Raman spectral fidelity, a strict linear concentration dependence, straightforward image interpretation, and quantification. [20][21][22][23][24][25][26][27][28] Recently, a new bioorthogonal imaging strategy is emerging, by introducing vibrational tags such as deuterium or alkyne labels to small biomolecules. [29][30][31][32] Through deuterium labeling in amino acids, glucose, cholesterol, and fatty acids, SRS microscopy was applied to visualize protein synthesis, 32 de novo lipogenesis, 33 intracellular cholesterol storage, 34 and metabolic activity in live tissues.…”

Section: Introductionmentioning

confidence: 99%

Bioorthogonal chemical imaging of metabolic changes during epithelial–mesenchymal transition of cancer cells by stimulated Raman scattering microscopy

Zhang

Min

2017

J. Biomed. Opt

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Study of metabolic changes during epithelial-mesenchymal transition (EMT) of cancer cells is important for basic understanding and therapeutic management of cancer progression. We here used metabolic labeling and stimulated Raman scattering (SRS) microscopy, a strategy of bioorthogonal chemical imaging, to directly visualize changes in anabolic metabolism during cancer EMT at a single-cell level. MCF-7 breast cancer cell is employed as a model system. Four types of metabolites (amino acids, glucose, fatty acids, and choline) are labeled with either deuterium or alkyne (C≡C) tag. Their intracellular incorporations into MCF-7 cells before or after EMT are visualized by SRS imaging targeted at the signature vibration frequency of C-D or C≡C bonds. Overall, after EMT, anabolism of amino acids, glucose, and choline is less active, reflecting slower protein and membrane synthesis in mesenchymal cells. Interestingly, we also observed less incorporation of glucose and palmitate acids into membrane lipids, but more of them into lipid droplets in mesenchymal cells. This result indicates that, although mesenchymal cells synthesize fewer membrane lipids, they are actively storing energy into lipid droplets, either through de novo lipogenesis from glucose or direct scavenging of exogenous free fatty acids. Hence, metabolic labeling coupled with SRS can be a straightforward method in imaging cancer metabolism.

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“…3 This technique results in intensity gain at the Stokes frequency, Stimulated Raman Gain (SRG), and intensity loss at the pump frequency, Stimulated Raman Loss (SRL). 4 Both the SRG and SRL signals can be orders of magnitude higher than the spontaneous Raman signal, and either may be used for molecular detection or imaging. 5,6 The increased stimulated Raman signal enables high sensitivity detection and high contrast imaging with fast integration times and low laser excitation powers, all of which are critical to biomedical applications.…”

Section: Introductionmentioning

confidence: 99%

Stimulated Raman with Broadband LED Stokes Source for Analysis of Glucose and Hemoglobin

Bullen

Kymissis

2018

Imaging and Applied Optics 2018 (3D, AO, AIO, COSI, DH, IS, LACSEA, LS&C, MATH, pcAOP)

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We demonstrate stimulated Raman gain using a broadband LED Stokes source to measure vibrational spectra of aqueous glucose solutions. This versatile and cost-effective method increases Raman signal for a variety of applications. We measured both stimulated Raman and spontaneous Raman spectra of glucose solutions with concentrations up to 10 mM with a photon counter and lock-in amplifier. We built partial least squares regression models based on both stimulated Raman and spontaneous Raman spectral data measured with each instrument for predicting concentrations of the glucose solutions. The stimulated Raman spectra measured with the lock-in amplifier based model had the strongest predictive power and predicted the concentrations of the test set of glucose solutions with a mean squared error value an order of magnitude lower than those of the spontaneous Raman based model.

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In Situ and In Vivo Molecular Analysis by Coherent Raman Scattering Microscopy

Cited by 44 publications

References 148 publications

Ultra-multiplex CARS spectroscopic imaging with 1-millisecond pixel dwell time

Ultra-multiplex CARS spectroscopic imaging with 1-millisecond pixel dwell time

Bioorthogonal chemical imaging of metabolic changes during epithelial–mesenchymal transition of cancer cells by stimulated Raman scattering microscopy

Stimulated Raman with Broadband LED Stokes Source for Analysis of Glucose and Hemoglobin

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