Hyperspectral Imaging with Stimulated Raman Scattering by Chirped Femtosecond Lasers

Fu, Dan; Holtom, Gary R.; Freudiger, Christian W.; Zhang, Xu; Xie, X. Sunney

doi:10.1021/jp308938t

Cited by 278 publications

(261 citation statements)

References 40 publications

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“…A large amount of work has been dedicated to access of vibrational spectral information at fast time rates, allowing differentiation of species in biological samples at sub-second time scales [7][8][9].…”

Section: Introductionmentioning

confidence: 99%

High-speed polarization-resolved coherent Raman scattering imaging

Hofer¹,

Balla²,

Brasselet³

2017

Optica

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Polarization-resolved coherent Raman scattering (polar-CRS) provides rich information on molecular orientational organization, with the strong advantages of being a label-free and chemically specific imaging method. Its implementation, however, strongly reduces the imaging acquisition rate, due to limits imposed by polarization tuning. Here we demonstrate fast-polar-CRS imaging based on combined electro-optic polarization and acousto-optic amplitude modulations, applicable to both stimulated Raman scattering and coherent anti-Stokes Raman scattering imaging. The proposed scheme adds polarization information without compromising the capacities of regular CRS intensity imaging; increases the speed of orientational imaging by two orders of magnitude as compared with previous approaches; and does not require post-processing analyses. We show that this method permits sub-second time-scale imaging of lipid order packing and local lipid membrane deformations in artificial lipid multilayers, but also in red blood cell ghosts, demonstrating its high sensitivity down to a single lipid bilayer membrane.

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Section: Introductionmentioning

confidence: 99%

High-speed polarization-resolved coherent Raman scattering imaging

Hofer¹,

Balla²,

Brasselet³

2017

Optica

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“…However, real-time identifying and mapping for multiple or possibly unknown molecules requires broad spectral breadth, desired resolution and high-speed acquisition, and has been a long-term pursuit in the field [5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23].…”

Section: Introductionmentioning

confidence: 99%

Spectral focusing dual-comb coherent anti-Stokes Raman spectroscopic imaging

Chen

et al. 2017

Opt. Lett.

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Coherent Raman microscopy provide label-free imaging by interrogating the intrinsic vibration of biomolecules.Nevertheless, trade-off between high chemical-specificity and high imaging-speed currently exists in transition from spectroscopy to spectroscopic imaging when capturing dynamics in complex living systems. Here, we present a novel concept in dual-comb scheme to substantially beat this trade-off and facilitate high-resolution broadband coherent antiStokes Raman spectroscopic imaging based on down-converted, automatically varying delay-time in spectral focusing excitation. A rapid measurement of vibrational microspectroscopy on sub-microsecond scale over a spectral span ~700 cm -1 with solid signal-to-noise ratio provides access to well-resolved molecular signatures within the fingerprint region. We demonstrate this high-performance spectroscopic imaging of spatially inhomogeneous distributions of chemical substances as well as the carotenoids accumulation in plant cells. This technique offers an unprecedented route for broadband CARS imaging with hundreds of kHz frame rate using available 1-GHz oscillators.

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“…However, YDFLs working at 990-1020 nm have rarely been demonstrated, mainly due to the small gain coefficient and the gain competition from the three-level and quasi-three-level bands [1]. Bridging this wavelength gap is crucial to achieve a wideband hyperspectral capability, which is essential in applications such as multicolor two-photon excited fluorescence (TPEF) microscopy [15] and hyperspectral coherent Raman scattering microscopy [16,17]. In addition, this special wavelength of 1010 nm by itself can potentially provide a higher TPEF efficiency for some widely-used biomolecules and dyes, e.g., Alexa Fluor 488 and emerald GFP, and thus reduce the illumination power requirement.…”

Section: Introductionmentioning

confidence: 99%

Compact fs ytterbium fiber laser at 1010 nm for biomedical applications

Kong

Pilger

Hachmeister

et al. 2017

Biomed. Opt. Express

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Ytterbium-doped fiber lasers (YDFLs) working in the near-infrared (NIR) spectral window and capable of high-power operation are popular in recent years. They have been broadly used in a variety of scientific and industrial research areas, including light bullet generation, optical frequency comb formation, materials fabrication, free-space laser communication, and biomedical diagnostics as well. The growing interest in YDFLs has also been cultivated for the generation of high-power femtosecond (fs) pulses. Unfortunately, the operating wavelengths of fs YDFLs have mostly been confined to two spectral bands, i.e., 970-980 nm through the three-level energy transition and 1030-1100 nm through the quasi three-level energy transition, leading to a spectral gap (990-1020 nm) in between, which is attributed to an intrinsically weak gain in this wavelength range. Here we demonstrate a highpower mode-locked fs YDFL operating at 1010 nm, which is accomplished in a compact and cost-effective package. It exhibits superior performance in terms of both short-term and longterm stability, i.e., <0.3% (peak intensity over 2.4 μs) and <4.0% (average power over 24 hours), respectively. To illustrate the practical applications, it is subsequently employed as a versatile fs laser for high-quality nonlinear imaging of biological samples, including twophoton excited fluorescence microscopy of mouse kidney and brain sections, as well as polarization-sensitive second-harmonic generation microscopy of potato starch granules and mouse tail muscle. It is anticipated that these efforts will largely extend the capability of fs YDFLs which is continuously tunable over 970-1100 nm wavelength range for wideband hyperspectral operations, serving as a promising complement to the gold-standard Ti:sapphire fs lasers.

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Hyperspectral Imaging with Stimulated Raman Scattering by Chirped Femtosecond Lasers

Cited by 278 publications

References 40 publications

High-speed polarization-resolved coherent Raman scattering imaging

High-speed polarization-resolved coherent Raman scattering imaging

Spectral focusing dual-comb coherent anti-Stokes Raman spectroscopic imaging

Compact fs ytterbium fiber laser at 1010 nm for biomedical applications

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