Abstract

Short laser pulse technology has significantly contributed to biomedical research, especially via nonlinear optical microscopy. Coherent anti-Stokes Raman scattering (CARS) microscopy is a label-free, chemical-selective method that is growing in importance as improved methods and light sources develop. Here we discuss different approaches to laser source development for CARS microscopy and highlight the advantages of a multimodal CARS microscope, illustrated by selected applications in biomedical research.

© 2010 Optical Society of America

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    [CrossRef] [PubMed]
  35. E. O. Potma, C. L. Evans, and X. S. Xie, “Heterodyne coherent anti-Stokes Raman scattering (CARS) imaging,” Opt. Lett. 31, 241–243 (2006).
    [CrossRef] [PubMed]
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    [CrossRef]
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    [CrossRef] [PubMed]
  45. E. O. Potma, W. P. de Boeij, P. J. M. van Haastert, and D. A. Wiersma, “Real-time visualization of intracellular hydrodynamics in single living cells,” Proc. Natl. Acad. Sci. USA 98, 1577–1582 (2001).
    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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  53. E. Bélanger, S. Bégin, S. Laffray, Y. D. Koninck, R. Vallée, and D. Côté, “Quantitative myelin imaging with coherent anti-Stokes Raman scattering microscopy: alleviating the excitation polarization dependence with circularly polarized laser beams,” Opt. Express 17, 18419–18432 (2009).
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2010

“Fluoview FV1000MPE femtocars add-on,” http://www.olympusamerica.com/seg_section/product.asp?product=1068&intCmp=seg_rdct_cars (2010).

2009

M. N. Slipchenko, T. T. Le, H. Chen, and J.-X. Cheng, “High-speed vibrational imaging and spectral analysis of lipid bodies by compound Raman microscopy,” J. Phys. Chem. B 113, 7681–7686 (2009).
[CrossRef] [PubMed]

R. K. Lyn, D. C. Kennedy, S. M. Sagan, D. R. Blais, Y. Rouleau, A. F. Pegoraro, X. S. Xie, A. Stolow, and J. P. Pezacki, “Direct imaging of the disruption of hepatitis c virus replication complexes by inhibitors of lipid metabolism,” Virology 394, 130–142 (2009).
[CrossRef] [PubMed]

F. P. Henry, D. Côté, M. A. Randolph, E. A. Z. Rust, R. W. Redmond, I. E. Kochevar, C. P. Lin, and J. M. Winograd, “Real-time in vivo assessment of the nerve microenvironment with coherent anti-Stokes Raman scattering microscopy,” Plast. Reconstr. Surg. 123, 123S–130S (2009).
[CrossRef] [PubMed]

A. F. Pegoraro, A. Ridsdale, D. J. Moffatt, Y. Jia, J. P. Pezacki, and A. Stolow, “Optimally chirped multimodal CARS microscopy based on a single Ti:sapphire oscillator,” Opt. Express 17, 2984–2996 (2009).
[CrossRef] [PubMed]

M. Cui, B. R. Bachler, and J. P. Ogilvie, “Comparing coherent and spontaneous Raman scattering under biological imaging conditions,” Opt. Lett. 34, 773–775 (2009).
[CrossRef] [PubMed]

K. Kieu, B. G. Saar, G. R. Holtom, X. S. Xie, and F. W. Wise, “High-power picosecond fiber source for coherent Raman microscopy,” Opt. Lett. 34, 2051–2053 (2009).
[CrossRef] [PubMed]

G. Krauss, T. Hanke, A. Sell, D. Träutlein, A. Leitenstorfer, R. Selm, M. Winterhalder, and A. Zumbusch, “Compact coherent anti-Stokes Raman scattering microscope based on a picosecond two-color Er:fiber laser system,” Opt. Lett. 34, 2847–2849 (2009).
[CrossRef] [PubMed]

E. Bélanger, S. Bégin, S. Laffray, Y. D. Koninck, R. Vallée, and D. Côté, “Quantitative myelin imaging with coherent anti-Stokes Raman scattering microscopy: alleviating the excitation polarization dependence with circularly polarized laser beams,” Opt. Express 17, 18419–18432 (2009).
[CrossRef]

A. F. Pegoraro, A. Ridsdale, D. J. Moffatt, J. P. Pezacki, B. K. Thomas, L. Fu, L. Dong, M. E. Fermann, and A. Stolow, “All-fiber CARS microscopy of live cells,” Opt. Express 17, 20700–20706 (2009).
[CrossRef] [PubMed]

2008

Y. Fu, T. B. Huff, H.-W. Wang, J.-X. Cheng, and H. Wang, “Ex vivo and in vivo imaging of myelin fibers in mouse brain by coherent anti-Stokes Raman scattering microscopy,” Opt. Express 16, 19396–19409 (2008).
[CrossRef] [PubMed]

C. P. Pfeffer, B. R. Olsen, F. Ganikhanov, and F. Légaré, “Multimodal nonlinear optical imaging of collagen arrays,” J. Struct. Biol. 164, 140–145 (2008).
[CrossRef] [PubMed]

I. Rocha-Mendoza, W. Langbein, and P. Borri, “Coherent anti-Stokes Raman microspectroscopy using spectral focusing with glass dispersion,” Appl. Phys. Lett. 93, 201103 (2008).
[CrossRef]

C. L. Evans and X. S. Xie, “Coherent anti-Stokes Raman scattering microscopy: chemical imaging for biology and medicine,” Annu. Rev. Anal. Chem. 1, 883–909 (2008).
[CrossRef]

2007

J.-X. Cheng, “Coherent anti-Stokes Raman scattering microscopy,” Appl. Spectrosc. 61, 197–208 (2007).
[CrossRef] [PubMed]

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J.-X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12, 054007 (2007).
[CrossRef] [PubMed]

F. Légaré, C. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J. 93, 1312–1320(2007).
[CrossRef] [PubMed]

H. A. Rinia, M. Bonn, M. Müller, and E. M. Vartiainen, “Quantitative CARS spectroscopy using the maximum entropy method: the main lipid phase transition,” Chem. PhysChem. 8, 279–287 (2007).
[CrossRef]

E. R. Andresen, C. K. Nielsen, J. Thøgersen, and S. R. Keiding, “Fiber laser-based light source for coherent anti-Stokes Raman scattering microspectroscopy,” Opt. Express 15, 4848–4856 (2007).
[CrossRef] [PubMed]

2006

X. Nan, A. M. Tonary, A. Stolow, X. S. Xie, and J. P. Pezacki, “Intracellular imaging of HCV RNA and cellular lipids by using simultaneous two-photon fluorescence and coherent anti-Stokes Raman scattering microscopies,” ChemBioChem 7, 1895–1897 (2006).
[CrossRef] [PubMed]

E. O. Potma, C. L. Evans, and X. S. Xie, “Heterodyne coherent anti-Stokes Raman scattering (CARS) imaging,” Opt. Lett. 31, 241–243 (2006).
[CrossRef] [PubMed]

J. P. Ogilvie, E. Beaurepaire, A. Alexandrou, and M. Joffre, “Fourier-transform coherent anti-Stokes Raman scattering microscopy,” Opt. Lett. 31, 480–482 (2006).
[CrossRef] [PubMed]

F. Ganikhanov, S. Carrasco, X. S. Xie, M. Katz, W. Seitz, and D. Kopf, “Broadly tunable dual-wavelength light source for coherent anti-Stokes Raman scattering microscopy,” Opt. Lett. 31, 1292–1294 (2006).
[CrossRef] [PubMed]

E. R. Andresen, V. Birkedal, J. Thøgersen, and S. R. Keiding, “Tunable light source for coherent anti-Stokes Raman scattering microspectroscopy based on the soliton self-frequency shift,” Opt. Lett. 31, 1328–1330 (2006).
[CrossRef] [PubMed]

Y. Fu, H. Wang, R. Shi, and J.-X. Cheng, “Characterization of photodamage in coherent anti-Stokes Raman scattering microscopy,” Opt. Express 14, 3942–3951 (2006).
[CrossRef] [PubMed]

F. Légaré, C. L. Evans, F. Ganikhanov, and X. S. Xie, “Towards CARS endoscopy,” Opt. Express 14, 4427–4432 (2006).
[CrossRef] [PubMed]

J.B.Pawley, ed., Handbook of Biological Confocal Microscopy (Springer, 2006).
[CrossRef]

2005

T. Betz, J. Teipel, D. Koch, W. Hartig, J. Guck, J. Kas, and H. Giessen, “Excitation beyond the monochromatic laser limit: simultaneous 3-D confocal and multiphoton microscopy with a tapered fiber as white-light laser source,” J. Biomed. Opt. 10, 054009 (2005).
[CrossRef] [PubMed]

A. Volkmer, “Vibrational imaging and microspectroscopies based on coherent anti-Stokes Raman scattering microscopy,” J. Phys. D 38, R59–R81 (2005).
[CrossRef]

C. L. Evans, E. O. Potma, M. Puoris’haag, D. Cote, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. USA 102, 16807–16812 (2005).
[CrossRef] [PubMed]

E. R. Andresen, H. N. Paulsen, V. Birkedal, J. Thøgersen, and S. R. Keiding, “Broadband multiplex coherent anti-Stokes Raman scattering microscopy employing photonic-crystal fibers,” J. Opt. Soc. Am. B 22, 1934–1938 (2005).
[CrossRef]

2004

J.-X. Cheng and X. Xie, “Coherent anti-Stokes Raman scattering microscopy: instrumentation, theory, and applications,” J. Phys. Chem. B 108, 827–840 (2004).
[CrossRef]

T. Hellerer, A. M. Enejder, and A. Zumbusch, “Spectral focusing: high spectral resolution spectroscopy with broad-bandwidth laser pulses,” Appl. Phys. Lett. 85, 25–27 (2004).
[CrossRef]

B. R. Masters, P. T. C. So, C. Buehler, N. Barry, J. D. Sutin, W. W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt. 9, 1265–1270 (2004).
[CrossRef] [PubMed]

2003

K. L. Corwin, N. R. Newbury, J. M. Dudley, S. Coen, S. A. Diddams, K. Weber, and R. S. Windeler, “Fundamental noise limitations to supercontinuum generation in microstructure fiber,” Phys. Rev. Lett. 90, 113904 (2003).
[CrossRef] [PubMed]

K. L. Corwin, N. R. Newbury, J. M. Dudley, S. Coen, S. A. Diddams, B. R. Washburn, K. Weber, and R. S. Windeller, “Fundamental amplitude noise limitations to supercontinuum spectra generated in a microstructured fiber,” Appl. Phys. B 77, 269–277 (2003).
[CrossRef]

M. Shiomi, T. Ito, S. Yamada, S. Kawashima, and J. Fan, “Development of an animal model for spontaneous myocardial infarction (WHHLMI Rabbit),” Arterioscler. Thromb. Vasc. Biol. 23, 1239–1244 (2003).
[CrossRef] [PubMed]

2002

J.-X. Cheng, Y. K. Jia, G. Zheng, and X. S. Xie, “Laser-scanning coherent anti-Stokes Raman scattering microscopy and applications to cell biology,” Biophys. J. 83, 502–509(2002).
[CrossRef] [PubMed]

N. Dudovich, D. Oron, and Y. Silberberg, “Single-pulse coherently controlled nonlinear Raman spectroscopy and microscopy,” Nature 418, 512–514 (2002).
[CrossRef] [PubMed]

M. Muller and J. M. Schins, “Imaging the thermodynamic state of lipid membranes with multiplex CARS microscopy,” J. Phys. Chem. B 106, 3715–3723 (2002).
[CrossRef]

J. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, “Multiplex coherent anti-Stokes Raman scattering microspectroscopy and study of lipid vesicles,” J. Phys. Chem. B 106, 8493–8498(2002).
[CrossRef]

D. J. Jones, E. O. Potma, J.-X. Cheng, B. Burfeindt, Y. Pang, J. Ye, and X. S. Xie, “Synchronization of two passively mode-locked, picosecond lasers within 20 fs for coherent anti-Stokes Raman scattering microscopy,” Rev. Sci. Instrum. 73, 2843–2848 (2002).
[CrossRef]

2001

A. Hopt and E. Neher, “Highly nonlinear photodamage in two-photon fluorescence microscopy,” Biophys. J. 80, 2029–2036(2001).
[CrossRef] [PubMed]

J.-x. Cheng, A. Volkmer, L. Book, and X. Xie, “An epi-detected coherent anti-Stokes Raman scattering (E-CARS) microscope with high spectral resolution and high sensitivity,” J. Phys. Chem. B 105, 1277–1280 (2001).
[CrossRef]

E. O. Potma, W. P. de Boeij, P. J. M. van Haastert, and D. A. Wiersma, “Real-time visualization of intracellular hydrodynamics in single living cells,” Proc. Natl. Acad. Sci. USA 98, 1577–1582 (2001).
[CrossRef] [PubMed]

1999

A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett. 82, 4142–4145 (1999).
[CrossRef]

P. J. Campagnola, M.-d. Wei, A. Lewis, and L. M. Loew, “High-resolution nonlinear optical imaging of live cells by second harmonic generation,” Biophys. J. 77, 3341–3349 (1999).
[CrossRef] [PubMed]

K. König, T. W. Becker, P. Fischer, I. Riemann, and K.-J. Halbhuber, “Pulse-length dependence of cellular response to intense near-infrared laser pulses in multiphoton microscopes,” Opt. Lett. 24, 113–115 (1999).
[CrossRef]

D. Yelin and Y. Silberberg, “Laser scanning third-harmonic-generation microscopy in biology,” Opt. Express 5, 169–175 (1999).
[CrossRef] [PubMed]

1994

1993

K. Duppen, F. de Haan, E. T. J. Nibbering, and D. A. Wiersma, “Chirped four-wave mixing,” Phys. Rev. A 47, 5120–5137(1993).
[CrossRef] [PubMed]

1992

E. T. J. Nibbering, D. A. Wiersma, and K. Duppen, “Ultrafast nonlinear spectroscopy with chirped optical pulses,” Phys. Rev. Lett. 68, 514–517 (1992).
[CrossRef] [PubMed]

1990

W. Denk, J. Strickler, and W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
[CrossRef] [PubMed]

1982

Alexandrou, A.

Andresen, E. R.

Bachler, B. R.

Barry, N.

B. R. Masters, P. T. C. So, C. Buehler, N. Barry, J. D. Sutin, W. W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt. 9, 1265–1270 (2004).
[CrossRef] [PubMed]

Beaurepaire, E.

Becker, T. W.

Bégin, S.

Bélanger, E.

Betz, T.

T. Betz, J. Teipel, D. Koch, W. Hartig, J. Guck, J. Kas, and H. Giessen, “Excitation beyond the monochromatic laser limit: simultaneous 3-D confocal and multiphoton microscopy with a tapered fiber as white-light laser source,” J. Biomed. Opt. 10, 054009 (2005).
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Blais, D. R.

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H. A. Rinia, M. Bonn, M. Müller, and E. M. Vartiainen, “Quantitative CARS spectroscopy using the maximum entropy method: the main lipid phase transition,” Chem. PhysChem. 8, 279–287 (2007).
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I. Rocha-Mendoza, W. Langbein, and P. Borri, “Coherent anti-Stokes Raman microspectroscopy using spectral focusing with glass dispersion,” Appl. Phys. Lett. 93, 201103 (2008).
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D. J. Jones, E. O. Potma, J.-X. Cheng, B. Burfeindt, Y. Pang, J. Ye, and X. S. Xie, “Synchronization of two passively mode-locked, picosecond lasers within 20 fs for coherent anti-Stokes Raman scattering microscopy,” Rev. Sci. Instrum. 73, 2843–2848 (2002).
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P. J. Campagnola, M.-d. Wei, A. Lewis, and L. M. Loew, “High-resolution nonlinear optical imaging of live cells by second harmonic generation,” Biophys. J. 77, 3341–3349 (1999).
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Chen, H.

M. N. Slipchenko, T. T. Le, H. Chen, and J.-X. Cheng, “High-speed vibrational imaging and spectral analysis of lipid bodies by compound Raman microscopy,” J. Phys. Chem. B 113, 7681–7686 (2009).
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Cheng, J.

J. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, “Multiplex coherent anti-Stokes Raman scattering microspectroscopy and study of lipid vesicles,” J. Phys. Chem. B 106, 8493–8498(2002).
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M. N. Slipchenko, T. T. Le, H. Chen, and J.-X. Cheng, “High-speed vibrational imaging and spectral analysis of lipid bodies by compound Raman microscopy,” J. Phys. Chem. B 113, 7681–7686 (2009).
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T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J.-X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12, 054007 (2007).
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Y. Fu, H. Wang, R. Shi, and J.-X. Cheng, “Characterization of photodamage in coherent anti-Stokes Raman scattering microscopy,” Opt. Express 14, 3942–3951 (2006).
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J.-X. Cheng, Y. K. Jia, G. Zheng, and X. S. Xie, “Laser-scanning coherent anti-Stokes Raman scattering microscopy and applications to cell biology,” Biophys. J. 83, 502–509(2002).
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D. J. Jones, E. O. Potma, J.-X. Cheng, B. Burfeindt, Y. Pang, J. Ye, and X. S. Xie, “Synchronization of two passively mode-locked, picosecond lasers within 20 fs for coherent anti-Stokes Raman scattering microscopy,” Rev. Sci. Instrum. 73, 2843–2848 (2002).
[CrossRef]

J.-x. Cheng, A. Volkmer, L. Book, and X. Xie, “An epi-detected coherent anti-Stokes Raman scattering (E-CARS) microscope with high spectral resolution and high sensitivity,” J. Phys. Chem. B 105, 1277–1280 (2001).
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C. L. Evans, E. O. Potma, M. Puoris’haag, D. Cote, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. USA 102, 16807–16812 (2005).
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K. L. Corwin, N. R. Newbury, J. M. Dudley, S. Coen, S. A. Diddams, B. R. Washburn, K. Weber, and R. S. Windeller, “Fundamental amplitude noise limitations to supercontinuum spectra generated in a microstructured fiber,” Appl. Phys. B 77, 269–277 (2003).
[CrossRef]

K. L. Corwin, N. R. Newbury, J. M. Dudley, S. Coen, S. A. Diddams, K. Weber, and R. S. Windeler, “Fundamental noise limitations to supercontinuum generation in microstructure fiber,” Phys. Rev. Lett. 90, 113904 (2003).
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Dong, L.

Dudley, J. M.

K. L. Corwin, N. R. Newbury, J. M. Dudley, S. Coen, S. A. Diddams, K. Weber, and R. S. Windeler, “Fundamental noise limitations to supercontinuum generation in microstructure fiber,” Phys. Rev. Lett. 90, 113904 (2003).
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K. L. Corwin, N. R. Newbury, J. M. Dudley, S. Coen, S. A. Diddams, B. R. Washburn, K. Weber, and R. S. Windeller, “Fundamental amplitude noise limitations to supercontinuum spectra generated in a microstructured fiber,” Appl. Phys. B 77, 269–277 (2003).
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N. Dudovich, D. Oron, and Y. Silberberg, “Single-pulse coherently controlled nonlinear Raman spectroscopy and microscopy,” Nature 418, 512–514 (2002).
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Duppen, K.

K. Duppen, F. de Haan, E. T. J. Nibbering, and D. A. Wiersma, “Chirped four-wave mixing,” Phys. Rev. A 47, 5120–5137(1993).
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E. T. J. Nibbering, D. A. Wiersma, and K. Duppen, “Ultrafast nonlinear spectroscopy with chirped optical pulses,” Phys. Rev. Lett. 68, 514–517 (1992).
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T. Hellerer, A. M. Enejder, and A. Zumbusch, “Spectral focusing: high spectral resolution spectroscopy with broad-bandwidth laser pulses,” Appl. Phys. Lett. 85, 25–27 (2004).
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F. Légaré, C. L. Evans, F. Ganikhanov, and X. S. Xie, “Towards CARS endoscopy,” Opt. Express 14, 4427–4432 (2006).
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E. O. Potma, C. L. Evans, and X. S. Xie, “Heterodyne coherent anti-Stokes Raman scattering (CARS) imaging,” Opt. Lett. 31, 241–243 (2006).
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C. L. Evans, E. O. Potma, M. Puoris’haag, D. Cote, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. USA 102, 16807–16812 (2005).
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B. R. Masters, P. T. C. So, C. Buehler, N. Barry, J. D. Sutin, W. W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt. 9, 1265–1270 (2004).
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T. Betz, J. Teipel, D. Koch, W. Hartig, J. Guck, J. Kas, and H. Giessen, “Excitation beyond the monochromatic laser limit: simultaneous 3-D confocal and multiphoton microscopy with a tapered fiber as white-light laser source,” J. Biomed. Opt. 10, 054009 (2005).
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T. Betz, J. Teipel, D. Koch, W. Hartig, J. Guck, J. Kas, and H. Giessen, “Excitation beyond the monochromatic laser limit: simultaneous 3-D confocal and multiphoton microscopy with a tapered fiber as white-light laser source,” J. Biomed. Opt. 10, 054009 (2005).
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Hellerer, T.

T. Hellerer, A. M. Enejder, and A. Zumbusch, “Spectral focusing: high spectral resolution spectroscopy with broad-bandwidth laser pulses,” Appl. Phys. Lett. 85, 25–27 (2004).
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F. P. Henry, D. Côté, M. A. Randolph, E. A. Z. Rust, R. W. Redmond, I. E. Kochevar, C. P. Lin, and J. M. Winograd, “Real-time in vivo assessment of the nerve microenvironment with coherent anti-Stokes Raman scattering microscopy,” Plast. Reconstr. Surg. 123, 123S–130S (2009).
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K. Kieu, B. G. Saar, G. R. Holtom, X. S. Xie, and F. W. Wise, “High-power picosecond fiber source for coherent Raman microscopy,” Opt. Lett. 34, 2051–2053 (2009).
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A. Hopt and E. Neher, “Highly nonlinear photodamage in two-photon fluorescence microscopy,” Biophys. J. 80, 2029–2036(2001).
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Ito, T.

M. Shiomi, T. Ito, S. Yamada, S. Kawashima, and J. Fan, “Development of an animal model for spontaneous myocardial infarction (WHHLMI Rabbit),” Arterioscler. Thromb. Vasc. Biol. 23, 1239–1244 (2003).
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Jia, Y.

Jia, Y. K.

J.-X. Cheng, Y. K. Jia, G. Zheng, and X. S. Xie, “Laser-scanning coherent anti-Stokes Raman scattering microscopy and applications to cell biology,” Biophys. J. 83, 502–509(2002).
[CrossRef] [PubMed]

Joffre, M.

Jones, D. J.

D. J. Jones, E. O. Potma, J.-X. Cheng, B. Burfeindt, Y. Pang, J. Ye, and X. S. Xie, “Synchronization of two passively mode-locked, picosecond lasers within 20 fs for coherent anti-Stokes Raman scattering microscopy,” Rev. Sci. Instrum. 73, 2843–2848 (2002).
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T. Betz, J. Teipel, D. Koch, W. Hartig, J. Guck, J. Kas, and H. Giessen, “Excitation beyond the monochromatic laser limit: simultaneous 3-D confocal and multiphoton microscopy with a tapered fiber as white-light laser source,” J. Biomed. Opt. 10, 054009 (2005).
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Kawashima, S.

M. Shiomi, T. Ito, S. Yamada, S. Kawashima, and J. Fan, “Development of an animal model for spontaneous myocardial infarction (WHHLMI Rabbit),” Arterioscler. Thromb. Vasc. Biol. 23, 1239–1244 (2003).
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R. K. Lyn, D. C. Kennedy, S. M. Sagan, D. R. Blais, Y. Rouleau, A. F. Pegoraro, X. S. Xie, A. Stolow, and J. P. Pezacki, “Direct imaging of the disruption of hepatitis c virus replication complexes by inhibitors of lipid metabolism,” Virology 394, 130–142 (2009).
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Kieu, K.

Koch, D.

T. Betz, J. Teipel, D. Koch, W. Hartig, J. Guck, J. Kas, and H. Giessen, “Excitation beyond the monochromatic laser limit: simultaneous 3-D confocal and multiphoton microscopy with a tapered fiber as white-light laser source,” J. Biomed. Opt. 10, 054009 (2005).
[CrossRef] [PubMed]

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F. P. Henry, D. Côté, M. A. Randolph, E. A. Z. Rust, R. W. Redmond, I. E. Kochevar, C. P. Lin, and J. M. Winograd, “Real-time in vivo assessment of the nerve microenvironment with coherent anti-Stokes Raman scattering microscopy,” Plast. Reconstr. Surg. 123, 123S–130S (2009).
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Koninck, Y. D.

Kopf, D.

Krauss, G.

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I. Rocha-Mendoza, W. Langbein, and P. Borri, “Coherent anti-Stokes Raman microspectroscopy using spectral focusing with glass dispersion,” Appl. Phys. Lett. 93, 201103 (2008).
[CrossRef]

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T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J.-X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12, 054007 (2007).
[CrossRef] [PubMed]

Le, T. T.

M. N. Slipchenko, T. T. Le, H. Chen, and J.-X. Cheng, “High-speed vibrational imaging and spectral analysis of lipid bodies by compound Raman microscopy,” J. Phys. Chem. B 113, 7681–7686 (2009).
[CrossRef] [PubMed]

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J.-X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12, 054007 (2007).
[CrossRef] [PubMed]

Légaré, F.

C. P. Pfeffer, B. R. Olsen, F. Ganikhanov, and F. Légaré, “Multimodal nonlinear optical imaging of collagen arrays,” J. Struct. Biol. 164, 140–145 (2008).
[CrossRef] [PubMed]

F. Légaré, C. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J. 93, 1312–1320(2007).
[CrossRef] [PubMed]

F. Légaré, C. L. Evans, F. Ganikhanov, and X. S. Xie, “Towards CARS endoscopy,” Opt. Express 14, 4427–4432 (2006).
[CrossRef] [PubMed]

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Lewis, A.

P. J. Campagnola, M.-d. Wei, A. Lewis, and L. M. Loew, “High-resolution nonlinear optical imaging of live cells by second harmonic generation,” Biophys. J. 77, 3341–3349 (1999).
[CrossRef] [PubMed]

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F. P. Henry, D. Côté, M. A. Randolph, E. A. Z. Rust, R. W. Redmond, I. E. Kochevar, C. P. Lin, and J. M. Winograd, “Real-time in vivo assessment of the nerve microenvironment with coherent anti-Stokes Raman scattering microscopy,” Plast. Reconstr. Surg. 123, 123S–130S (2009).
[CrossRef] [PubMed]

C. L. Evans, E. O. Potma, M. Puoris’haag, D. Cote, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. USA 102, 16807–16812 (2005).
[CrossRef] [PubMed]

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T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J.-X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12, 054007 (2007).
[CrossRef] [PubMed]

Loew, L. M.

P. J. Campagnola, M.-d. Wei, A. Lewis, and L. M. Loew, “High-resolution nonlinear optical imaging of live cells by second harmonic generation,” Biophys. J. 77, 3341–3349 (1999).
[CrossRef] [PubMed]

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R. K. Lyn, D. C. Kennedy, S. M. Sagan, D. R. Blais, Y. Rouleau, A. F. Pegoraro, X. S. Xie, A. Stolow, and J. P. Pezacki, “Direct imaging of the disruption of hepatitis c virus replication complexes by inhibitors of lipid metabolism,” Virology 394, 130–142 (2009).
[CrossRef] [PubMed]

Mantulin, W. W.

B. R. Masters, P. T. C. So, C. Buehler, N. Barry, J. D. Sutin, W. W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt. 9, 1265–1270 (2004).
[CrossRef] [PubMed]

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Masters, B. R.

B. R. Masters, P. T. C. So, C. Buehler, N. Barry, J. D. Sutin, W. W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt. 9, 1265–1270 (2004).
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M. Muller and J. M. Schins, “Imaging the thermodynamic state of lipid membranes with multiplex CARS microscopy,” J. Phys. Chem. B 106, 3715–3723 (2002).
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H. A. Rinia, M. Bonn, M. Müller, and E. M. Vartiainen, “Quantitative CARS spectroscopy using the maximum entropy method: the main lipid phase transition,” Chem. PhysChem. 8, 279–287 (2007).
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X. Nan, A. M. Tonary, A. Stolow, X. S. Xie, and J. P. Pezacki, “Intracellular imaging of HCV RNA and cellular lipids by using simultaneous two-photon fluorescence and coherent anti-Stokes Raman scattering microscopies,” ChemBioChem 7, 1895–1897 (2006).
[CrossRef] [PubMed]

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A. Hopt and E. Neher, “Highly nonlinear photodamage in two-photon fluorescence microscopy,” Biophys. J. 80, 2029–2036(2001).
[CrossRef] [PubMed]

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K. L. Corwin, N. R. Newbury, J. M. Dudley, S. Coen, S. A. Diddams, B. R. Washburn, K. Weber, and R. S. Windeller, “Fundamental amplitude noise limitations to supercontinuum spectra generated in a microstructured fiber,” Appl. Phys. B 77, 269–277 (2003).
[CrossRef]

K. L. Corwin, N. R. Newbury, J. M. Dudley, S. Coen, S. A. Diddams, K. Weber, and R. S. Windeler, “Fundamental noise limitations to supercontinuum generation in microstructure fiber,” Phys. Rev. Lett. 90, 113904 (2003).
[CrossRef] [PubMed]

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K. Duppen, F. de Haan, E. T. J. Nibbering, and D. A. Wiersma, “Chirped four-wave mixing,” Phys. Rev. A 47, 5120–5137(1993).
[CrossRef] [PubMed]

E. T. J. Nibbering, D. A. Wiersma, and K. Duppen, “Ultrafast nonlinear spectroscopy with chirped optical pulses,” Phys. Rev. Lett. 68, 514–517 (1992).
[CrossRef] [PubMed]

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Ogilvie, J. P.

Olsen, B. R.

C. P. Pfeffer, B. R. Olsen, F. Ganikhanov, and F. Légaré, “Multimodal nonlinear optical imaging of collagen arrays,” J. Struct. Biol. 164, 140–145 (2008).
[CrossRef] [PubMed]

F. Légaré, C. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J. 93, 1312–1320(2007).
[CrossRef] [PubMed]

Oron, D.

N. Dudovich, D. Oron, and Y. Silberberg, “Single-pulse coherently controlled nonlinear Raman spectroscopy and microscopy,” Nature 418, 512–514 (2002).
[CrossRef] [PubMed]

Pang, Y.

D. J. Jones, E. O. Potma, J.-X. Cheng, B. Burfeindt, Y. Pang, J. Ye, and X. S. Xie, “Synchronization of two passively mode-locked, picosecond lasers within 20 fs for coherent anti-Stokes Raman scattering microscopy,” Rev. Sci. Instrum. 73, 2843–2848 (2002).
[CrossRef]

Paulsen, H. N.

Pegoraro, A. F.

Pezacki, J. P.

A. F. Pegoraro, A. Ridsdale, D. J. Moffatt, J. P. Pezacki, B. K. Thomas, L. Fu, L. Dong, M. E. Fermann, and A. Stolow, “All-fiber CARS microscopy of live cells,” Opt. Express 17, 20700–20706 (2009).
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R. K. Lyn, D. C. Kennedy, S. M. Sagan, D. R. Blais, Y. Rouleau, A. F. Pegoraro, X. S. Xie, A. Stolow, and J. P. Pezacki, “Direct imaging of the disruption of hepatitis c virus replication complexes by inhibitors of lipid metabolism,” Virology 394, 130–142 (2009).
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A. F. Pegoraro, A. Ridsdale, D. J. Moffatt, Y. Jia, J. P. Pezacki, and A. Stolow, “Optimally chirped multimodal CARS microscopy based on a single Ti:sapphire oscillator,” Opt. Express 17, 2984–2996 (2009).
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X. Nan, A. M. Tonary, A. Stolow, X. S. Xie, and J. P. Pezacki, “Intracellular imaging of HCV RNA and cellular lipids by using simultaneous two-photon fluorescence and coherent anti-Stokes Raman scattering microscopies,” ChemBioChem 7, 1895–1897 (2006).
[CrossRef] [PubMed]

Pfeffer, C.

F. Légaré, C. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J. 93, 1312–1320(2007).
[CrossRef] [PubMed]

Pfeffer, C. P.

C. P. Pfeffer, B. R. Olsen, F. Ganikhanov, and F. Légaré, “Multimodal nonlinear optical imaging of collagen arrays,” J. Struct. Biol. 164, 140–145 (2008).
[CrossRef] [PubMed]

Potma, E. O.

E. O. Potma, C. L. Evans, and X. S. Xie, “Heterodyne coherent anti-Stokes Raman scattering (CARS) imaging,” Opt. Lett. 31, 241–243 (2006).
[CrossRef] [PubMed]

C. L. Evans, E. O. Potma, M. Puoris’haag, D. Cote, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. USA 102, 16807–16812 (2005).
[CrossRef] [PubMed]

D. J. Jones, E. O. Potma, J.-X. Cheng, B. Burfeindt, Y. Pang, J. Ye, and X. S. Xie, “Synchronization of two passively mode-locked, picosecond lasers within 20 fs for coherent anti-Stokes Raman scattering microscopy,” Rev. Sci. Instrum. 73, 2843–2848 (2002).
[CrossRef]

E. O. Potma, W. P. de Boeij, P. J. M. van Haastert, and D. A. Wiersma, “Real-time visualization of intracellular hydrodynamics in single living cells,” Proc. Natl. Acad. Sci. USA 98, 1577–1582 (2001).
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C. L. Evans, E. O. Potma, M. Puoris’haag, D. Cote, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. USA 102, 16807–16812 (2005).
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F. P. Henry, D. Côté, M. A. Randolph, E. A. Z. Rust, R. W. Redmond, I. E. Kochevar, C. P. Lin, and J. M. Winograd, “Real-time in vivo assessment of the nerve microenvironment with coherent anti-Stokes Raman scattering microscopy,” Plast. Reconstr. Surg. 123, 123S–130S (2009).
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F. P. Henry, D. Côté, M. A. Randolph, E. A. Z. Rust, R. W. Redmond, I. E. Kochevar, C. P. Lin, and J. M. Winograd, “Real-time in vivo assessment of the nerve microenvironment with coherent anti-Stokes Raman scattering microscopy,” Plast. Reconstr. Surg. 123, 123S–130S (2009).
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Ridsdale, A.

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H. A. Rinia, M. Bonn, M. Müller, and E. M. Vartiainen, “Quantitative CARS spectroscopy using the maximum entropy method: the main lipid phase transition,” Chem. PhysChem. 8, 279–287 (2007).
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I. Rocha-Mendoza, W. Langbein, and P. Borri, “Coherent anti-Stokes Raman microspectroscopy using spectral focusing with glass dispersion,” Appl. Phys. Lett. 93, 201103 (2008).
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R. K. Lyn, D. C. Kennedy, S. M. Sagan, D. R. Blais, Y. Rouleau, A. F. Pegoraro, X. S. Xie, A. Stolow, and J. P. Pezacki, “Direct imaging of the disruption of hepatitis c virus replication complexes by inhibitors of lipid metabolism,” Virology 394, 130–142 (2009).
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F. P. Henry, D. Côté, M. A. Randolph, E. A. Z. Rust, R. W. Redmond, I. E. Kochevar, C. P. Lin, and J. M. Winograd, “Real-time in vivo assessment of the nerve microenvironment with coherent anti-Stokes Raman scattering microscopy,” Plast. Reconstr. Surg. 123, 123S–130S (2009).
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M. Muller and J. M. Schins, “Imaging the thermodynamic state of lipid membranes with multiplex CARS microscopy,” J. Phys. Chem. B 106, 3715–3723 (2002).
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N. Dudovich, D. Oron, and Y. Silberberg, “Single-pulse coherently controlled nonlinear Raman spectroscopy and microscopy,” Nature 418, 512–514 (2002).
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A. F. Pegoraro, A. Ridsdale, D. J. Moffatt, Y. Jia, J. P. Pezacki, and A. Stolow, “Optimally chirped multimodal CARS microscopy based on a single Ti:sapphire oscillator,” Opt. Express 17, 2984–2996 (2009).
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X. Nan, A. M. Tonary, A. Stolow, X. S. Xie, and J. P. Pezacki, “Intracellular imaging of HCV RNA and cellular lipids by using simultaneous two-photon fluorescence and coherent anti-Stokes Raman scattering microscopies,” ChemBioChem 7, 1895–1897 (2006).
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B. R. Masters, P. T. C. So, C. Buehler, N. Barry, J. D. Sutin, W. W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt. 9, 1265–1270 (2004).
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X. Nan, A. M. Tonary, A. Stolow, X. S. Xie, and J. P. Pezacki, “Intracellular imaging of HCV RNA and cellular lipids by using simultaneous two-photon fluorescence and coherent anti-Stokes Raman scattering microscopies,” ChemBioChem 7, 1895–1897 (2006).
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E. O. Potma, W. P. de Boeij, P. J. M. van Haastert, and D. A. Wiersma, “Real-time visualization of intracellular hydrodynamics in single living cells,” Proc. Natl. Acad. Sci. USA 98, 1577–1582 (2001).
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H. A. Rinia, M. Bonn, M. Müller, and E. M. Vartiainen, “Quantitative CARS spectroscopy using the maximum entropy method: the main lipid phase transition,” Chem. PhysChem. 8, 279–287 (2007).
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C. L. Evans and X. S. Xie, “Coherent anti-Stokes Raman scattering microscopy: chemical imaging for biology and medicine,” Annu. Rev. Anal. Chem. 1, 883–909 (2008).
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F. Légaré, C. L. Evans, F. Ganikhanov, and X. S. Xie, “Towards CARS endoscopy,” Opt. Express 14, 4427–4432 (2006).
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C. L. Evans, E. O. Potma, M. Puoris’haag, D. Cote, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. USA 102, 16807–16812 (2005).
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D. J. Jones, E. O. Potma, J.-X. Cheng, B. Burfeindt, Y. Pang, J. Ye, and X. S. Xie, “Synchronization of two passively mode-locked, picosecond lasers within 20 fs for coherent anti-Stokes Raman scattering microscopy,” Rev. Sci. Instrum. 73, 2843–2848 (2002).
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J. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, “Multiplex coherent anti-Stokes Raman scattering microspectroscopy and study of lipid vesicles,” J. Phys. Chem. B 106, 8493–8498(2002).
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J.-X. Cheng, Y. K. Jia, G. Zheng, and X. S. Xie, “Laser-scanning coherent anti-Stokes Raman scattering microscopy and applications to cell biology,” Biophys. J. 83, 502–509(2002).
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D. J. Jones, E. O. Potma, J.-X. Cheng, B. Burfeindt, Y. Pang, J. Ye, and X. S. Xie, “Synchronization of two passively mode-locked, picosecond lasers within 20 fs for coherent anti-Stokes Raman scattering microscopy,” Rev. Sci. Instrum. 73, 2843–2848 (2002).
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Zheng, G.

J.-X. Cheng, Y. K. Jia, G. Zheng, and X. S. Xie, “Laser-scanning coherent anti-Stokes Raman scattering microscopy and applications to cell biology,” Biophys. J. 83, 502–509(2002).
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G. Krauss, T. Hanke, A. Sell, D. Träutlein, A. Leitenstorfer, R. Selm, M. Winterhalder, and A. Zumbusch, “Compact coherent anti-Stokes Raman scattering microscope based on a picosecond two-color Er:fiber laser system,” Opt. Lett. 34, 2847–2849 (2009).
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A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett. 82, 4142–4145 (1999).
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Annu. Rev. Anal. Chem.

C. L. Evans and X. S. Xie, “Coherent anti-Stokes Raman scattering microscopy: chemical imaging for biology and medicine,” Annu. Rev. Anal. Chem. 1, 883–909 (2008).
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Appl. Phys. B

K. L. Corwin, N. R. Newbury, J. M. Dudley, S. Coen, S. A. Diddams, B. R. Washburn, K. Weber, and R. S. Windeller, “Fundamental amplitude noise limitations to supercontinuum spectra generated in a microstructured fiber,” Appl. Phys. B 77, 269–277 (2003).
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Appl. Phys. Lett.

T. Hellerer, A. M. Enejder, and A. Zumbusch, “Spectral focusing: high spectral resolution spectroscopy with broad-bandwidth laser pulses,” Appl. Phys. Lett. 85, 25–27 (2004).
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Biophys. J.

J.-X. Cheng, Y. K. Jia, G. Zheng, and X. S. Xie, “Laser-scanning coherent anti-Stokes Raman scattering microscopy and applications to cell biology,” Biophys. J. 83, 502–509(2002).
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Chem. PhysChem.

H. A. Rinia, M. Bonn, M. Müller, and E. M. Vartiainen, “Quantitative CARS spectroscopy using the maximum entropy method: the main lipid phase transition,” Chem. PhysChem. 8, 279–287 (2007).
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ChemBioChem

X. Nan, A. M. Tonary, A. Stolow, X. S. Xie, and J. P. Pezacki, “Intracellular imaging of HCV RNA and cellular lipids by using simultaneous two-photon fluorescence and coherent anti-Stokes Raman scattering microscopies,” ChemBioChem 7, 1895–1897 (2006).
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J. Biomed. Opt.

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J.-X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12, 054007 (2007).
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B. R. Masters, P. T. C. So, C. Buehler, N. Barry, J. D. Sutin, W. W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt. 9, 1265–1270 (2004).
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J. Opt. Soc. Am. B

J. Phys. Chem. B

J.-X. Cheng and X. Xie, “Coherent anti-Stokes Raman scattering microscopy: instrumentation, theory, and applications,” J. Phys. Chem. B 108, 827–840 (2004).
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J.-x. Cheng, A. Volkmer, L. Book, and X. Xie, “An epi-detected coherent anti-Stokes Raman scattering (E-CARS) microscope with high spectral resolution and high sensitivity,” J. Phys. Chem. B 105, 1277–1280 (2001).
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M. N. Slipchenko, T. T. Le, H. Chen, and J.-X. Cheng, “High-speed vibrational imaging and spectral analysis of lipid bodies by compound Raman microscopy,” J. Phys. Chem. B 113, 7681–7686 (2009).
[CrossRef] [PubMed]

M. Muller and J. M. Schins, “Imaging the thermodynamic state of lipid membranes with multiplex CARS microscopy,” J. Phys. Chem. B 106, 3715–3723 (2002).
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J. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, “Multiplex coherent anti-Stokes Raman scattering microspectroscopy and study of lipid vesicles,” J. Phys. Chem. B 106, 8493–8498(2002).
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Nature

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Opt. Express

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A. F. Pegoraro, A. Ridsdale, D. J. Moffatt, J. P. Pezacki, B. K. Thomas, L. Fu, L. Dong, M. E. Fermann, and A. Stolow, “All-fiber CARS microscopy of live cells,” Opt. Express 17, 20700–20706 (2009).
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Y. Fu, H. Wang, R. Shi, and J.-X. Cheng, “Characterization of photodamage in coherent anti-Stokes Raman scattering microscopy,” Opt. Express 14, 3942–3951 (2006).
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F. Légaré, C. L. Evans, F. Ganikhanov, and X. S. Xie, “Towards CARS endoscopy,” Opt. Express 14, 4427–4432 (2006).
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E. R. Andresen, C. K. Nielsen, J. Thøgersen, and S. R. Keiding, “Fiber laser-based light source for coherent anti-Stokes Raman scattering microspectroscopy,” Opt. Express 15, 4848–4856 (2007).
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Y. Fu, T. B. Huff, H.-W. Wang, J.-X. Cheng, and H. Wang, “Ex vivo and in vivo imaging of myelin fibers in mouse brain by coherent anti-Stokes Raman scattering microscopy,” Opt. Express 16, 19396–19409 (2008).
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A. F. Pegoraro, A. Ridsdale, D. J. Moffatt, Y. Jia, J. P. Pezacki, and A. Stolow, “Optimally chirped multimodal CARS microscopy based on a single Ti:sapphire oscillator,” Opt. Express 17, 2984–2996 (2009).
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Opt. Lett.

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G. Krauss, T. Hanke, A. Sell, D. Träutlein, A. Leitenstorfer, R. Selm, M. Winterhalder, and A. Zumbusch, “Compact coherent anti-Stokes Raman scattering microscope based on a picosecond two-color Er:fiber laser system,” Opt. Lett. 34, 2847–2849 (2009).
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Figures (6)

Fig. 1
Fig. 1

(a) Energy level diagram for resonant CARS. The energy difference between pump ω p and Stokes ω s matches the energy of the Raman level to be probed Ω R . The anti-Stokes ω a s light is the desired signal. (b) Energy level diagrams for nonresonant CARS. The anti-Stokes light generated has the same frequency as is generated in the resonant case, yet does not rely on ground-state vibrational coherence.

Fig. 2
Fig. 2

The CARS (a) resonant signal, (b) contrast, and (c) performance are shown as a function of pump-Stokes frequency difference for varying pulse spectral widths for pulses of constant energy. The broadest (i.e., temporally shortest) pulse achieves the maximum resonant signal whereas maximum contrast is achieved for the narrowest (i.e., temporally longest) pulse. The individual peaks of the Raman spectrum are resolved only by use of the narrowest pulse. Performance is maximized for a pulse spectral width that matches the effective linewidth of the combined Raman mode; see text for definitions of contrast and performance.

Fig. 3
Fig. 3

Generalized CARS microscopy optical setup. The pump laser is split using a beam splitter (BS). Part of the beam is used to generate a Stokes pulse; the other part is delayed then recombined and overlapped with the Stokes on a dichroic mirror (DM). Inside the microscope galvo-scanning mirrors (GS) direct the beam. Dichroic mirrors and bandpass (BP) filters separate the signals from the excitation light. Epidetected signals can be collected before or after the galvo mirrors (descanned or nondescanned detection).

Fig. 4
Fig. 4

(a) CARS and DIC multimodal imaging of HuH-7 liver cells. The CARS image was taken first using chirped fs pulses and then overlapped with a subsequent DIC image. In print, some of the lipid drops are indicated; online, red represents CARS due to lipids, gray represents the DIC image. Scale bar is 5 μm . (b) CARS and TPF multimodal image of HuH-7 cells infected with a modified HCV strain (infected cells express GFP). The images were acquired simultaneously using chirped fs pulses as part of a time course studying live cells (taken at the 48 h time point). In print, the areas indicated are TPF due to GFP; the remainder of the image is CARS due to lipids. Online, greenrepresents TPF due to GFP; red represents CARS due to lipids. Scale bar is 10 μm .

Fig. 5
Fig. 5

(a) WHHMLI arterial sample. Online, red represents CARS imaging of lipids, green represents TPF from elastin, and blue represents SHG from collagen. In print, the different parts of the sample are labeled. All the signals were collected simultaneously using chirped fs pulses. Scale bar is 50 μm . Multimodal nonlinear optical imaging of fascia. (b) Forward SHG image using fs pulses, (c) forward CARS image at Raman shift frequency Δ ω = 2845 cm 1 using ps pulses, and (d) forward THG image using fs pulses. Scanning area was 146.2 μm 2 ( 512 × 512 pixels). (b), (c) and (d) reprinted with permission from [50].

Fig. 6
Fig. 6

In vivo CARS imaging using ps pulses of mouse brain using an upright microscope with a dipping mode water objective. (a) A schematic of the in vivo CARS microscope; D, dichroic mirror. (b) CARS image of the parietal cortex. (c) Epi-CARS image of bundles of myelinated fibers in the subcortex white matter. Reprinted with permission from [52].

Equations (3)

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P CARS ( ω a s ) = + χ ( 3 ) E p ( ω p ) E s * ( ω s ) E p r ( ω p r ) d ω ,
χ r ( 3 ) = A Ω ( ω p ω s ) i Γ ,
E ( t ) = E π δ 2 ln 2 exp ( π 2 δ 2 t 2 2 ln 2 ) exp ( i ω t ) ,

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