Chemically sensitive bioimaging with coherent Raman scattering

Raman spectroscopy is an increasingly popular technique in many areas including biology and medicine.It is based on Raman scattering, a phenomenon in which incident photons lose or gain energy via interactions with vibrating molecules in a sample. These energy shifts can be used to obtain information regarding molecular composition of the sample with very high accuracy. Applications of Raman spectroscopy in the life sciences have included quantification of biomolecules, hyperspectral molecular imaging of cells and tissue, medical diagnosis, and others. This review briefly presents the physical origin of Raman scattering explaining the key classical and quantum mechanical concepts. Variations of the Raman effect will also be considered, including resonance, coherent, and enhanced Raman scattering. We discuss the molecular origins of prominent bands often found in the Raman spectra of biological samples. Finally, we examine several variations of Raman spectroscopy techniques in practice, looking at their applications, strengths, and challenges. This review is intended to be a starting resource for scientists new to Raman spectroscopy, providing theoretical background and practical examples as the foundation for further study and exploration.

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“…Many other approaches have also been developed, and are discussed in more detail in the recent review by Camp and Cicerone [21].…”

Section: Typical Cars Systemmentioning

confidence: 99%

“…This increase in signal and imaging speed comes at the expense of diverse spectral information, often acquiring signal at only one or a few wavenumbers. [20][21][22][23]. The low Raman scattering signal also limits the technique's ability to detect small concentrations of molecules.…”

Section: Introductionmentioning

confidence: 99%

Raman spectroscopy: techniques and applications in the life sciences

Shipp¹,

Sinjab²,

Notingher³

2017

Adv. Opt. Photon.

268

189

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“…In order to study dynamic processes, such as enzymatic hydrolysis of cellulose, imaging speed is increased at the expense of spectral information, i.e. single-vibration coherent Raman scattering techniques can achieve video-rate imaging speed, while image with full Raman fingerprint information in each pixel requires acquisition times on the minutes scale (Camp and Cicerone 2015). Other imaging modalities can be implemented together with coherent Raman scattering to form a multimodal nonlinear optical microscope.…”

Section: Introductionmentioning

confidence: 99%

Visualization of structural changes in cellulosic substrates during enzymatic hydrolysis using multimodal nonlinear microscopy

et al. 2016

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Enzymatic hydrolysis of cellulose provides a renewable source of monosaccharides for production of variety of biochemicals and biopolymers. Unfortunately, the enzymatic hydrolysis of cellulose is often incomplete, and the reasons are not fully understood. We have monitored enzymatic hydrolysis in terms of molecular density, ordering and autofluorescence of cellulose structures in real time using simultaneous CARS, SHG and MPEF microscopy with the aim of contributing to the understanding and optimization of the enzymatic hydrolysis of cellulose. Three celluloserich substrates with different supramolecular structures, pulp fibre, acid-treated pulp fibre and Avicel, were studied at microscopic level. The microscopy studies revealed that before enzymatic hydrolysis Avicel had the greatest carbon-hydrogen density, while pulp fibre and acid-treated fibre had similar density. Monitoring of the substrates during enzymatic hydrolysis revealed the double exponential SHG decay for pulp fibre and acid-treated fibre indicating two phases of the process. Acid-treated fibre was hydrolysed most rapidly and the hydrolysis of pulp fibre was spatially non-uniform leading to fractioning of the particles, while the hydrolysis of Avicel was more than an order of magnitude slower than that of both fibres.

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“…CARS occurs when at least two light pulses-pump/probe and Stokes, having respective frequencies of ω P,Pr and ω S -interact in a non-linear material such that the difference frequency, Ω R = ω P,Pr -ω S , matches a vibrational mode of the material, resulting in the resonant generation of photons at the anti-Stokes frequency ω AS = 2ω P,Pr -ω S [1][2][3]. With its combination of rapid imaging with rich spectroscopic information, CARS "hyper-microscopy" is gaining popularity in research fields such as biomedical imaging [4][5][6], materials science [7], and biology [8][9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Spectrally-broad coherent anti-Stokes Raman scattering hyper-microscopy utilizing a Stokes supercontinuum pumped at 800 nm

Porquez

Cole

Tabarangao

et al. 2016

Biomed. Opt. Express

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Abstract:We demonstrate spectral-focusing based coherent anti-Stokes Raman scattering (SF-CARS) hyper-microscopy capable of probing vibrational frequencies from 630 cm 1 to 3250 cm 1 using a single Ti:Sapphire femtosecond laser operating at 800 nm, and a commercially-available supercontinuum-generating fibre module. A broad Stokes supercontinuum with significant spectral power at wavelengths between 800 nm and 940 nm is generated by power tuning the fibre module using atypically long and/or chirped ~200 fs pump pulses, allowing convenient access to lower vibrational frequencies in the fingerprint spectral region. This work significantly reduces the instrumental and technical requirements for multimodal CARS microscopy, while expanding the spectral capabilities of an established approach to SF-CARS.

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Chemically sensitive bioimaging with coherent Raman scattering

Cited by 412 publications

References 113 publications

Raman spectroscopy: techniques and applications in the life sciences

Raman spectroscopy: techniques and applications in the life sciences

Visualization of structural changes in cellulosic substrates during enzymatic hydrolysis using multimodal nonlinear microscopy

Spectrally-broad coherent anti-Stokes Raman scattering hyper-microscopy utilizing a Stokes supercontinuum pumped at 800 nm

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