A multimodal platform for nonlinear optical microscopy and microspectroscopy

Chen, Hongtao; Wang, Haifeng; Slipchenko, Mikhail N.; Jung, Yookyung; Shi, Yunzhou; Zhu, Jiabin; Buhman, Kimberly K.; Cheng, Ji‐Xin

doi:10.1364/oe.17.001282

Cited by 113 publications

(101 citation statements)

References 35 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%

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

Yue

Cheng

2010

Journal of Lipid Research

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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%

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

Yue

Cheng

2010

Journal of Lipid Research

Self Cite

147

128

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“…SHG and THG have the advantage of being nontoxic since no energy is absorbed by the specimen during imaging (Fig. 2), and they can be combined with MPM providing with the opportunity to perform multimodal imaging (Campagnola and Loew, 2003;Chen et al, 2009;Debarre et al, 2006;Farrar et al, 2011;Radosevich et al, 2008).…”

Section: Second and Third Harmonic Generation (Shg And Thg)mentioning

confidence: 99%

Salivary Glands: A Powerful Experimental System to Study Cell Biology in Live Animals by Intravital Microscopy

Šrámková¹,

Porat-Shliom²,

Wigand³

et al. 2012

Current Frontiers and Perspectives in Cell Biology

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“…In addition, NLO microscopy is sensitive to specific molecules and structures [1][2][3][4][5][6]. Recently, multimodal NLO microscopy has been extended to include two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG), and coherence anti-Stokes Raman scattering (CARS) imaging on the same platform [7][8][9]. Multimodal NLO microscopy allows simultaneous visualization of different endogenous and exogenous structures and specific molecules in complex tissue samples.…”

Section: Introductionmentioning

confidence: 99%

“…In previous multimodal NLO microscopy papers, a single picosecond (ps) pulsed laser system, a single femtosecond (fs) pulsed laser system with a photonic crystal fiber (PCF), or multi-laser systems with a flip mirror for switching have been used [7][8][9][10][11][12]. However, in this study, we constructed a multimodal NLO microscope using lights that were combined of an fs-pulsed laser and a ps-pulsed laser incident into a sample, simultaneously.…”

Section: Introductionmentioning

confidence: 99%

Multimodal Nonlinear Optical Microscopy for Simultaneous 3-D Label-Free and Immunofluorescence Imaging of Biological Samples

Park

Lee

et al. 2014

Journal of the Optical Society of Korea

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In this study, we demonstrated multimodal nonlinear optical (NLO) microscopy integrated simultaneously with two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG), and coherent anti-Stokes Raman scattering (CARS) in order to obtain targeted cellular and label-free images in an immunofluorescence assay of the atherosclerotic aorta from apolipoprotein E-deficient mice. The multimodal NLO microscope used two laser systems: picosecond (ps) and femtosecond (fs) pulsed lasers. A pair of ps-pulsed lights served for CARS (817 nm and 1064 nm) and SHG (817 nm) images; light from the fs-pulsed laser with the center wavelength of 720 nm was incident into the sample to obtain autofluorescence and targeted molecular TPEF images for high efficiency of fluorescence intensity without cross-talk. For multicolor-targeted TPEF imaging, we stained smooth-muscle cells and macrophages with fluorescent dyes (Alexa Fluor 350 and Alexa Fluor 594) for an immunofluorescence assay. Each depth-sectioned image consisted of 512 × 512 pixels with a field of view of 250 × 250 µm 2 , a lateral resolution of 0.4 µm, and an axial resolution of 1.3 µm. We obtained composite multicolor images with conventional label-free NLO images and targeted TPEF images in atherosclerotic-plaque samples. Multicolor 3-D imaging of atherosclerotic-plaque structural and functional composition will be helpful for understanding the pathogenesis of cardiovascular disease.

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A multimodal platform for nonlinear optical microscopy and microspectroscopy

Cited by 113 publications

References 35 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

Salivary Glands: A Powerful Experimental System to Study Cell Biology in Live Animals by Intravital Microscopy

Multimodal Nonlinear Optical Microscopy for Simultaneous 3-D Label-Free and Immunofluorescence Imaging of Biological Samples

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