All-fiber CARS microscopy of live cells

Pegoraro, Adrian F.; Ridsdale, Andrew; Moffatt, Douglas J.; Pezacki, John Paul; Thomas, Brian K.; Fu, Libin; Dong, Liang; Fermann, M. E.; Stolow, Albert

doi:10.1364/oe.17.020700

Cited by 74 publications

(52 citation statements)

References 25 publications

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“…Therefore, toward the goal of coupling the CARS imaging modality to other widely used platforms of multiphoton microscopy, Pegoraro et al ( 38 ) and Chen et al ( 39 ) showed independently high-speed CARS imaging of lipids with femtosecond (fs) laser pulses. Commercial CARS microscopes with either ps or fs laser sources are now available through Leica and Olympus, respectively.…”

Section: Coupling Cars Imaging With Spontaneous Raman Spectroscopymentioning

confidence: 99%

“…Furthermore, the integration of spontaneous Raman spectroscopy onto the CARS microscope platform provides complete high-resolution Raman spectral information of the samples ( 47 ). Compact laser sources ( 38,117,118 ) are being developed to make CARS microscopy more accessible to biomedical researchers by reducing its cost, footprint, and complexity. Increasing the penetration depth with CARS endoscopy ( 84,85 ) or miniature microscope objective ( 119 ) could allow CARS imaging from the lumen of the intestine, esophagus, and artery, or CARS imaging of lipid-rich tissues with minimally invasive surgery.…”

Section: Study Of Myelin Diseasesmentioning

confidence: 99%

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Shedding new light on lipid biology with coherent anti-Stokes Raman scattering microscopy

Yue

Cheng

2010

Journal of Lipid Research

148

130

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Despite the ubiquitous roles of lipids in biology, the detection of lipids has relied on invasive techniques and population measurements. The molecular profi les of the lipid-rich structures are evaluated by measurements of total lipid extracts with liquid or gas chromatography coupled to mass spectrometry ( 8 ). This approach is inexact because it measures lipid molecules from areas other than the structures of interest. Furthermore, spatial information, which is critical to understanding the function of lipids ( 9 ), is inaccessible with the bulk measurement techniques. To visualize the lipid-rich structures, fl uorescently tagged lipid molecules, which intercalate with the lipidrich structures, are used ( 10, 11 ). Nevertheless, the use of the fl uorescent lipid molecules to label lipid-rich structures is problematic for several reasons. First, to facilitate the transport of the fl uorescent lipid molecules into cells, cell fi xation procedures are often performed ( 12 ). This procedure prevents monitoring the dynamic responses of lipid-rich structures to stimuli or the disease processes. Second, labeling effi ciency is highly dependent on the type of fl uorescent lipid molecules ( 12 ), thus posing a signifi cant problem to the quantitation of lipid-rich structures. Third, the intercalation of fl uorescent lipid molecules can lead to changes in the properties of lipidrich structures. For instance, incorporation of fl uorescent lipid molecules into the cell membrane can induce membrane phase separation and membrane microdomain formation, which can perturb critical cellular processes including signal transduction and endocytosis ( 13 ).As an alternative to fl uorescence, signals from molecular vibration provide an attractive means for label-free chemical imaging. In particular, Raman scattering has been widely used for spectroscopic study of biomolecules ( 14 ). The Raman-scattered photons exhibit frequency Abstract Despite the ubiquitous roles of lipids in biology, the detection of lipids has relied on invasive techniques, population measurements, or nonspecifi c labeling. Such diffi culties can be circumvented by a label-free imaging technique known as coherent anti-Stokes Raman (CARS) microscopy, which is capable of chemically selective, highly sensitive, and high-speed imaging of lipid-rich structures with submicron three-dimensional spatial resolution. We review the broad applications of CARS microscopy to studies of lipid biology in cell cultures, tissue biopsies, and model organisms. Recent technical advances, limitations of the technique, and perspectives are discussed. Lipids play a critical role in human health and diseases. Phospholipids, glycolipids, and sterol lipids are the major components of the cell membrane ( 1 ). Sphingolipids constitute up to 80% of the myelin sheaths, which are the membranous structures that wrap around the axons for insulation and for proper propagation of electrical impulses ( 2 ). Glycerolipids or triglycerides serve as the cytoplasmic energy depots ( 3 ). Bioactive lip...

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Section: Coupling Cars Imaging With Spontaneous Raman Spectroscopymentioning

confidence: 99%

Section: Study Of Myelin Diseasesmentioning

confidence: 99%

Shedding new light on lipid biology with coherent anti-Stokes Raman scattering microscopy

Yue

Cheng

2010

Journal of Lipid Research

148

130

View full text Add to dashboard Cite

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“…Attenuated peaks were observed at 2870, 2910 and 2950 cm -1 , which were attributed to lipid phase changes and changes in protein content. Pegoraro et al (2009b) recently demonstrated a novel spectral focusing implementation to acquire high resolution CARS spectra in the C--H stretching vibration region, using an optimally chirped multimodal CARS microscopy based on a single femtosecond Ti:sapphire laser and synchronized PCF output. Preliminary data showed that clear spectral differences could be detected from multiple regions of interest identified in a rabbit artery section.…”

Section: Broadband Cars Imagingmentioning

confidence: 99%

Nonlinear optical microscopy in decoding arterial diseases

Ridsdale

Mostaço-Guidolin

et al. 2012

Biophys Rev

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Pathological understanding of arterial diseases is mainly attributable to histological observations based on conventional tissue staining protocols. The emerging development of nonlinear optical microscopy (NLOM), particularly in second-harmonic generation, two-photon excited fluorescence and coherent Raman scattering, provides a new venue to visualize pathological changes in the extracellular matrix caused by atherosclerosis progression. These techniques in general require minimal tissue preparation and offer rapid three-dimensional imaging. The capability of label-free microscopic imaging enables disease impact to be studied directly on the bulk artery tissue, thus minimally perturbing the sample. In this review, we look at recent progress in applications related to arterial disease imaging using various forms of NLOM.

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“…With the improved power distribution, an enhanced CARS signal can be achieved when using PCF 5 i.e. a faster imaging process than achieved with other PCF based CARS systems (4 µs/pixel) [16]. Note that the commercially available fiber is not polarization maintaining.…”

Section: Optimum Pcf For Carsmentioning

confidence: 99%

“…Previously, CARS setups using PCF for generation of the Stokes wavelength have been presented [15,16]. However, wavelength conversion to the relevant IR wavelength was weak.…”

Section: Introductionmentioning

confidence: 99%

Supercontinuum generation for coherent anti-Stokes Raman scattering microscopy with photonic crystal fibers

Klarskov¹,

Isomäki²,

Hansen³

et al. 2011

Opt. Express

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Photonic crystal fiber (PCF) designs with two zero-dispersion wavelengths (ZDWs) are experimentally investigated in order to suggest a novel PCF for coherent anti-Stokes Raman scattering (CARS) microscopy. From our investigation, we select the optimum PCF design and demonstrate a tailored spectrum with power concentrated around the relevant wavelengths for lipid imaging (648 nm and 1027 nm). This new PCF is characterized by varying the fiber length, the average power, and the pulse width of the fs pump pulses. It was found that the selected PCF design gave a significantly improved spectral distribution compared to an existing PCF for CARS microscopy. Furthermore, the PCF is designed in a twofold symmetric structure allowing for polarization maintaining propagation. Finally, the pulse propagation is investigated numerically showing good agreement with the measured spectrum. From the numerical analysis, the nonlinear effects responsible for the spectral broadening are explained to be soliton fission processes, dispersive waves, and stimulated Raman scattering. ©2011 Optical Society of America

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All-fiber CARS microscopy of live cells

Cited by 74 publications

References 25 publications

Shedding new light on lipid biology with coherent anti-Stokes Raman scattering microscopy

Shedding new light on lipid biology with coherent anti-Stokes Raman scattering microscopy

Nonlinear optical microscopy in decoding arterial diseases

Supercontinuum generation for coherent anti-Stokes Raman scattering microscopy with photonic crystal fibers

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