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Optics Express

Optics Express

  • Editor: Michael Duncan
  • Vol. 14, Iss. 7 — Apr. 3, 2006
  • pp: 2798–2804
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In-vivo multi-nonlinear optical imaging of a living cell using a supercontinuum light source generated from a photonic crystal fiber

Hideaki Kano and Hiro-o Hamaguchi  »View Author Affiliations


Optics Express, Vol. 14, Issue 7, pp. 2798-2804 (2006)
http://dx.doi.org/10.1364/OE.14.002798


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Abstract

A supercontinuum light source generated with a femtosecond Ti:Sapphire oscillator has been used to obtain both vibrational and two-photon excitation fluorescence (TPEF) images of a living cell simultaneously at different wavelengths. Owing to an ultrabroadband spectral profile of the supercontinuum, multiple vibrational resonances have been detected through coherent anti-Stokes Raman scattering (CARS) process. In addition to the multiplex CARS process, multiple electronic states can be excited due to the broadband electronic two-photon excitation using the supercontinuum, giving rise to a two-photon excitation fluorescence (TPEF) signal. Using a living yeast cell whose nucleus is labeled by green fluorescent protein (GFP), we have succeeded in visualizing organelles such as mitochondria, septum, and nucleus through the CARS and the TPEF processes. The supercontinuum enables us to perform unique multi-nonlinear optical imaging through two different nonlinear optical processes.

© 2006 Optical Society of America

1. Introduction

Raman microspectroscopy is one of the most powerful and nondestructive methods in order to elucidate intra-cellular structure and its dynamics in vivo with three-dimensional sectioning capability [1–8

1. G. J. Puppels, F. F. M. De Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, “Studying single living cells and chromosomes by confocal Raman microspectroscopy,” Nature (London, United Kingdom) 347, 301–303 (1990). [CrossRef] [PubMed]

]. Based on the time- and space-resolved molecular specific information obtained by confocal Raman microspectroscopy, we have recently investigated the mitochondrial metabolic activity and an initial death process of a living yeast cell [5–8

5. Y.-S. Huang, T. Karashima, M. Yamamoto, and H. Hamaguchi, “Molecular-level pursuit of yeast mitosis by time- and space-resolved Raman spectroscopy,” J. Raman Spectrosc. 34, 1–3 (2003). [CrossRef]

].The “Raman spectroscopic signature of life” has been found in living fission [5–7

5. Y.-S. Huang, T. Karashima, M. Yamamoto, and H. Hamaguchi, “Molecular-level pursuit of yeast mitosis by time- and space-resolved Raman spectroscopy,” J. Raman Spectrosc. 34, 1–3 (2003). [CrossRef]

] and budding [8

8. Y. Naito, A. Toh-e, and H.-o. Hamaguchi, “In vivo time-resolved Raman imaging of a spontaneous death process of a single budding yeast cell,” J. Raman Spectrosc. 36, 837–839 (2005). [CrossRef]

] yeast cells, enabling quantitative analysis of cellular bioactivity at the molecular level. In order to trace the detailed dynamical behavior, however, spontaneous Raman microspectroscopy may not be suitable because of its low efficiency; it often takes several minutes to obtain one spectrum. This low efficiency originates from the small scattering cross section of the spontaneous Raman process. An alternative approach to obtain vibrational images with high speed is coherent Raman microspectroscopy. Among them, coherent anti-Stokes Raman scattering (CARS) microscopy has been widely exploited [9–15

9. 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]

]. In particular, multiplex CARS microspectroscopy is promising because of its capability to obtain vibrational spectra efficiently [12

12. G. W. H. Wurpel, J. M. Schins, and M. Mueller, “Chemical specificity in three-dimensional imaging with multiplex coherent anti-Stokes Raman scattering microscopy,” Opt. Lett. 27, 1093–1095 (2002). [CrossRef]

, 16–19

16. C. Otto, A. Voroshilov, S. G. Kruglik, and J. Greve, “Vibrational bands of luminescent zinc(II)-octaethyl-porphyrin using a polarization-sensitive “microscopic” multiplex CARS technique,” J. Raman Spectrosc. 32, 495–501 (2001). [CrossRef]

]. The multiplex CARS process requires two laser sources, namely, a narrow band pump laser (ω1) and a broadband Stokes laser (ω2). The multiple vibrational coherences are created because of the wide spectral range of the frequency difference, ω12. If we can prepare ultrashort laser pulses, an impulsive Raman excitation and a subsequent narrow-band probe can also generate a multiplex CARS spectrum [20

20. H. Kano and H. Hamaguchi, “Dispersion-compensated supercontinuum generation for ultrabroadband multiplex coherent anti-Stokes Raman scattering spectroscopy,” J. Raman Spectrosc., in press.

, 21

21. S.-H. Lim, A. G. Caster, and S. R. Leone, “Single-pulse phase-control interferometric coherent anti-Stokes Raman scattering spectroscopy,” Phys. Rev. A 30, 2805–2807 (2005).

]. One of the most prominent features of multiplex CARS microspectroscopy lies in the fact that it can easily distinguish the concentration change of a particular molecule from the structural change through the spectral analysis. It should be emphasized that a single-wavenumber CARS detection, which is widely adopted in CARS microscopy, cannot discriminate these two phenomena. Although there were several restrictions on the spectral coverage of multiplex CARS microspectroscopy mainly due to the bandwidth of the laser emission [12

12. G. W. H. Wurpel, J. M. Schins, and M. Mueller, “Chemical specificity in three-dimensional imaging with multiplex coherent anti-Stokes Raman scattering microscopy,” Opt. Lett. 27, 1093–1095 (2002). [CrossRef]

, 16–19

16. C. Otto, A. Voroshilov, S. G. Kruglik, and J. Greve, “Vibrational bands of luminescent zinc(II)-octaethyl-porphyrin using a polarization-sensitive “microscopic” multiplex CARS technique,” J. Raman Spectrosc. 32, 495–501 (2001). [CrossRef]

], the spectral coverage has been significantly broadened using the supercontinuum light source generated from a photonic crystal fiber [22–25

22. I. G. Petrov and V. V. Yakovlev, “Enhancing red-shifted white-light continuum generation in optical fibers for applications in nonlinear Raman microscopy,” Opt. Express 13, 1299 (2005)http://www.opticsexpress.org/abstract.cfm?URI=OPEX-13-4-1299 . [CrossRef] [PubMed]

] or a tapered fiber [26

26. T. W. Kee and M. T. Cicerone, “Simple approach to one-laser, broadband coherent anti-Stokes Raman scattering microscopy,” Opt. Lett. 29, 2701–2703 (2004). [CrossRef] [PubMed]

]. Recently, the spectral coverage of the multiplex CARS microspectroscopy has been extended to be more than 2800 cm-1 [20

20. H. Kano and H. Hamaguchi, “Dispersion-compensated supercontinuum generation for ultrabroadband multiplex coherent anti-Stokes Raman scattering spectroscopy,” J. Raman Spectrosc., in press.

], which is ranging from 360 nm to 3210 cm-1. In view of electronic spectroscopy, the supercontinuum light source can also be used as an excitation light source for the two-photon excitation fluorescence (TPEF) [27–29

27. C. McConnell and E. Riis, “Photonic crystal fibre enables short-wavelength two-photon laser scanning fluorescence microscopy with fura-2,” Phys. Med. Biol. 49, 4757–4763 (2004). [CrossRef] [PubMed]

]. Owing to the broadband spectral profile of the supercontinuum, the two-photon allowed electronic state can be excited efficiently in comparison with conventional TPEF microscopy using a Ti:Sapphire oscillator. In the present study, we have combined our multiplex CARS setup with the TPEF detection. Both CARS and TPEF signals have been successfully obtained simultaneously within short data-acquisition time such as 100ms.

2. Experimental

The supercontinuum-based nonlinear optical microspectroscopy system has been described elsewhere [24

24. H. Kano and H. Hamaguchi, “Ultrabroadband (>2500 cm-1) Multiplex coherent anti-stokes raman scattering microspectroscopy using a supercontinuum generated from a photonic crystal fiber,” Appl. Phys. Lett. 86, 121113–121115 (2005). [CrossRef]

]. Briefly, an unamplified femtosecond mode-locked Ti:Sapphire oscillator (Coherent, Vitesse-800) was used as a laser source. Typical duration, pulse energy, peak wavelength, and repetition rate were 100 fs, 12 nJ, 800 nm, and 80 MHz, respectively. About 20 % of the output from the oscillator was used for a seed laser to generate a supercontinuum in the PCF (Crystal Fibre, NL-PM-750). The fundamental of the Ti:Sapphire laser and the supercontinuum were used for the pump (ω1) and Stokes (ω2) lasers, respectively. In order to obtain CARS spectrum with high frequency resolution, the pump laser pulses were spectrally filtered using a narrow band pass filter. The bandwidth of the pump laser is about 20cm-1. The pulse energy of the pump and Stokes lasers were 200 and 170 pJ, respectively. Two laser pulses were superimposed collinearly using an 800-nm Notch filter, and then tightly focused onto the sample with a microscope objective (x40). We have modified an inverted microscope (Nikon, TE2000-S). The forward-propagating CARS signal was collected with another microscope objective (x40) in an opposed configuration. After passing through an 800-nm Notch and short wavelength pass filters, the CARS signal was spectrally dispersed by a polychromator (Acton, SpectraPro-300i) and detected by a CCD camera (Roper Scientific, Spec-10:400BR/XTE). The sample was scanned by a piezo stage (MadCity, Nano-LP-100). An exposure time for each point was 100 ms. The spatial resolution was estimated to be 0.47 ± 0.01μm for the lateral and 1.51± 0.02μm for the axial directions, respectively [30

30. H. Kano and H. Hamaguchi, “Vibrational imaging of a J-aggregate microcrystal using ultrabroadband multiplex coherent anti-Stokes Raman scattering microspectroscopy,” submitted to Vibrational Spectroscopy.

]. We used fission yeast Schizosaccharomyces pombe (S. pombe) as a sample [5–7

5. Y.-S. Huang, T. Karashima, M. Yamamoto, and H. Hamaguchi, “Molecular-level pursuit of yeast mitosis by time- and space-resolved Raman spectroscopy,” J. Raman Spectrosc. 34, 1–3 (2003). [CrossRef]

]. The nuclei of yeast cells were labeled by green fluorescent protein (GFP). Yeast cells in water were spread on a slide-glass and sandwiched with a cover-glass. Because of a small quantity of the sample, yeast cells were immobilized between a slide-glass and a cover-glass. All measurements were performed at room temperature.

3. Results and Discussion

Fig. 1. (a) Spectral profiles of the CARS signal of a living yeast cell (red) and surrounding water (blue) obtained in 100 ms exposure time; (b) CARS image of living yeast cell at a Raman shift of 2840 cm-1. The CARS signals at the positions of black and white crosses correspond to the red and blue curves in Fig. 1(a), respectively.

Thanks to the three-dimensional sectioning capability, CARS microscopy enables us to obtain not only a lateral but also an axial slice of a living yeast cell. Figures 2 (a) and 2(b) show a lateral and an axial CARS images of a yeast cell, respectively. Figure 2(b) corresponds to the vertical slice of the yeast cell at the position of y=0. The CARS signal is weaker at the top part rather than the bottom part. It is due to the imperfect focusing of the two laser beams because of the spatially heterogeneous refractive index inside of the cell. Figure 2(c) shows axial slices of the same yeast cell at each depth position. Three-dimensional distribution of mitochondoria is clearly observed in Fig. 2(c) with the high three-dimensional spatial resolution.

Fig. 2. Lateral(a) and axial(b) CARS images of a yeast cell at a Raman shift of 2840 cm-1; (c) Axial slices of the yeast cell at different depth positions, which are indicated at the top in a micrometer scale.

Figure 4 shows multi-nonlinear optical imaging of the CARS (red) and TPEF (green) signals, which is obtained in Fig. 3. As described, the yeast cell at the center of Fig. 4 is in the M phase. It is also found that the mitochondria exist around the nucleus.

Fig. 3. Spectral profile of the CARS and TPEF signals of a living yeast cell; CARS lateral images of living yeast cells at the Raman shift of 2840 cm-1 at the delay time of zero (b) and -4 ps (c), respectively; TPEF lateral images of the same system at 506 nm at the delay time of zero (d) and -4 ps (e), respectively; TPEF image only due to the non-degenerated (ω12)- photon process.
Fig. 4. In-vivo multi-nonlinear optical imaging of yeast cells. The CARS and TPEF images are indicated as red and green, respectively.

4. Conclusion

In conclusion, multi-nonlinear optical imaging of a living yeast cell has been performed by supercontinuum-based microspectroscopy. Owing to the broadband feature of the supercontinuum, the spectral profile of the multiplex CARS signal can be elucidated in detail. In addition to the ultrabroadband multiplex CARS detection, the efficient two-photon excitation can also be performed in resonance with the two-photon allowed electronic state. Since the Raman signal due to a nucleus is weak to be detected, dual imaging of the CARS and the TPEF signals provides useful information on the dynamical behavior of the living system with high speed. The supercontinuum enables us to perform unique multi-nonlinear optical imaging through various nonlinear optical processes.

Acknowledgments

This research is supported by a Grant-in-Aid for Creative Scientific Research (No. 15GS0204) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan. H. K. is supported by a Grant-in-Aid for Young Scientists (B) (No. 15750005) from Japan Society for the Promotion of Science, and research grants from The Kurata Memorial Hitachi Science and Technology Foundation. The authors thank Dr. Y. -S. Huang for her help in sample preparation and Dr. T. Karashima and Prof. M. Yamamoto for supplying us the strain of S. pombe.

References and links

1.

G. J. Puppels, F. F. M. De Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, “Studying single living cells and chromosomes by confocal Raman microspectroscopy,” Nature (London, United Kingdom) 347, 301–303 (1990). [CrossRef] [PubMed]

2.

G. J. Puppels, J. H. Olminkhof, G. M. Segers-Nolten, C. Otto, F. F. de Mul, and J. Greve, “Laser irradiation and Raman spectroscopy of single living cells and chromosomes: sample degradation occurs with 514.5 nm but not with 660 nm laser light,” Exp. Cell Res. 195, 361–367 (1991). [CrossRef] [PubMed]

3.

Y. Takai, T. Masuko, and H. Takeuchi, “Lipid structure of cytotoxic granules in living human killer T lymphocytes studied by Raman microspectroscopy,” Biochim. Biophys. Acta 1335, 199–208 (1997). [CrossRef] [PubMed]

4.

C. Otto, N. M. Sijtsema, and J. Greve, “Confocal Raman microspectroscopy of the activation of single neutrophilic granulocytes,” Eur. Biophys. J. 27, 582–589 (1998). [CrossRef] [PubMed]

5.

Y.-S. Huang, T. Karashima, M. Yamamoto, and H. Hamaguchi, “Molecular-level pursuit of yeast mitosis by time- and space-resolved Raman spectroscopy,” J. Raman Spectrosc. 34, 1–3 (2003). [CrossRef]

6.

Y.-S. Huang, T. Karashima, M. Yamamoto, T. Ogura, and H. Hamaguchi, “Raman spectroscopic signature of life in a living yeast cell,” J. Raman Spectrosc. 35, 525–526 (2004). [CrossRef]

7.

Y.-S. Huang, T. Karashima, M. Yamamoto, and H. Hamaguchi, “Molecular-level investigation of the structure, transformation, and bioactivity of single living fission yeast cells by time- and space-resolved Raman Spectroscopy,” Biochemistry 44, 10009–10019 (2005). [CrossRef] [PubMed]

8.

Y. Naito, A. Toh-e, and H.-o. Hamaguchi, “In vivo time-resolved Raman imaging of a spontaneous death process of a single budding yeast cell,” J. Raman Spectrosc. 36, 837–839 (2005). [CrossRef]

9.

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]

10.

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]

11.

M. Hashimoto, T. Araki, and S. Kawata, “Molecular vibration imaging in the fingerprint region by use of coherent anti-Stokes Raman scattering microscopy with a collinear configuration,” Opt. Lett. 25, 1768–1770 (2000). [CrossRef]

12.

G. W. H. Wurpel, J. M. Schins, and M. Mueller, “Chemical specificity in three-dimensional imaging with multiplex coherent anti-Stokes Raman scattering microscopy,” Opt. Lett. 27, 1093–1095 (2002). [CrossRef]

13.

H. N. Paulsen, K. M. Hilligsoe, J. Thogersen, S. R. Keiding, and J. J. Larsen, “Coherent anti-Stokes Raman scattering microscopy with a photonic crystal fiber based light source,” Opt. Lett. 28, 1123–1125 (2003). [CrossRef] [PubMed]

14.

R. D. Schaller, J. Ziegelbauer, L. F. Lee, L. H. Haber, and R. J. Saykally, “Chemically selective imaging of subcellular structure in human hepatocytes with coherent anti-stokes Raman scattering (CARS) near-field scanning optical microscopy (NSOM),” J. Phys. Chem. B 106, 8489–8492 (2002). [CrossRef]

15.

T. Ichimura, N. Hayazawa, M. Hashimoto, Y. Inouye, and S. Kawata, “Tip-enhanced coherent anti-stokes Raman scattering for vibrational nanoimaging,” Phys. Rev. Lett. 92, 220801 (2004). [CrossRef] [PubMed]

16.

C. Otto, A. Voroshilov, S. G. Kruglik, and J. Greve, “Vibrational bands of luminescent zinc(II)-octaethyl-porphyrin using a polarization-sensitive “microscopic” multiplex CARS technique,” J. Raman Spectrosc. 32, 495–501 (2001). [CrossRef]

17.

J.-X. 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]

18.

D. Oron, N. Dudovich, and Y. Silberberg, “Single-pulse phase-contrast nonlinear Raman Spectroscopy,” Phys. Rev. Lett. 89, 273001 (2002). [CrossRef]

19.

D. Oron, N. Dudovich, D. Yelin, and Y. Silberberg, “Narrow-band coherent anti-stokes Raman signals from broad-band pulses,” Phys. Rev. Lett. 88, 063004 (2002). [CrossRef] [PubMed]

20.

H. Kano and H. Hamaguchi, “Dispersion-compensated supercontinuum generation for ultrabroadband multiplex coherent anti-Stokes Raman scattering spectroscopy,” J. Raman Spectrosc., in press.

21.

S.-H. Lim, A. G. Caster, and S. R. Leone, “Single-pulse phase-control interferometric coherent anti-Stokes Raman scattering spectroscopy,” Phys. Rev. A 30, 2805–2807 (2005).

22.

I. G. Petrov and V. V. Yakovlev, “Enhancing red-shifted white-light continuum generation in optical fibers for applications in nonlinear Raman microscopy,” Opt. Express 13, 1299 (2005)http://www.opticsexpress.org/abstract.cfm?URI=OPEX-13-4-1299 . [CrossRef] [PubMed]

23.

S. O. Konorov, D. A. Akimov, E. E. Serebryannikov, A. A. Ivanov, M. V. Alfimov, and A. M. Zheltikov, “Cross-correlation frequency-resolved optical gating coherent anti-Stokes Raman scattering with frequency-converting photonic-crystal fibers,” Phys. Rev. E 70, 057601 (2004). [CrossRef]

24.

H. Kano and H. Hamaguchi, “Ultrabroadband (>2500 cm-1) Multiplex coherent anti-stokes raman scattering microspectroscopy using a supercontinuum generated from a photonic crystal fiber,” Appl. Phys. Lett. 86, 121113–121115 (2005). [CrossRef]

25.

H. Kano and H. Hamaguchi, “Vibrationally resonant imaging of a single living cell by supercontinuum-based multiplex coherent anti-Stokes Raman scattering microspectroscopy,” Opt. Express 13, 1322–1327 (2005)http://www.opticsexpress.org/abstract.cfm?URI=OPEX-13-4-1322 . [CrossRef] [PubMed]

26.

T. W. Kee and M. T. Cicerone, “Simple approach to one-laser, broadband coherent anti-Stokes Raman scattering microscopy,” Opt. Lett. 29, 2701–2703 (2004). [CrossRef] [PubMed]

27.

C. McConnell and E. Riis, “Photonic crystal fibre enables short-wavelength two-photon laser scanning fluorescence microscopy with fura-2,” Phys. Med. Biol. 49, 4757–4763 (2004). [CrossRef] [PubMed]

28.

K. Isobe, W. Watanabe, S. Matsunaga, T. Higashi, K. Fukui, and K. Itoh, “Multi-spectral two-photon excited fluorescence microscopy using supercontinuum light source,” Jpn. J. Appl. Phys. Part 2 44, L167–L169 (2005). [CrossRef]

29.

J. A. Palero, V. O. Boer, J. C. Vijverberg, H. C. Gerritsen, and H. J. C. M. Sterenborg, “Short-wavelength two-photon excitation fluorescence microscopy of tryptophan with a photonic crystal fiber based light source,” Opt. Express 13, 5363–5368 (2005)http://www.opticsexpress.org/abstract.cfm?URI=OPEX-13-14-5363.. [CrossRef] [PubMed]

30.

H. Kano and H. Hamaguchi, “Vibrational imaging of a J-aggregate microcrystal using ultrabroadband multiplex coherent anti-Stokes Raman scattering microspectroscopy,” submitted to Vibrational Spectroscopy.

31.

H. Wang, Y. Fu, P. Zickmund, R. Shi, and J.-X. Cheng, “Coherent anti-Stokes Raman scattering imaging of live spinal tissues,” Biophys. J. 89, 581–591 (2005). [CrossRef] [PubMed]

OCIS Codes
(110.0180) Imaging systems : Microscopy
(180.2520) Microscopy : Fluorescence microscopy
(190.4180) Nonlinear optics : Multiphoton processes
(190.4370) Nonlinear optics : Nonlinear optics, fibers
(300.6230) Spectroscopy : Spectroscopy, coherent anti-Stokes Raman scattering

ToC Category:
Nonlinear Optics

History
Original Manuscript: January 3, 2006
Manuscript Accepted: March 15, 2006
Published: April 3, 2006

Virtual Issues
Vol. 1, Iss. 5 Virtual Journal for Biomedical Optics

Citation
Hideaki Kano and Hiro-o Hamaguchi, "In-vivo multi-nonlinear optical imaging of a living cell using a supercontinuum light source generated from a photonic crystal fiber," Opt. Express 14, 2798-2804 (2006)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-14-7-2798


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References

  1. G. J. Puppels, F. F. M. De Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, "Studying single living cells and chromosomes by confocal Raman microspectroscopy," Nature (London, United Kingdom) 347,301-303 (1990). [CrossRef] [PubMed]
  2. G. J. Puppels, J. H. Olminkhof, G. M. Segers-Nolten, C. Otto, F. F. de Mul, and J. Greve, "Laser irradiation and Raman spectroscopy of single living cells and chromosomes: sample degradation occurs with 514.5 nm but not with 660 nm laser light," Exp. Cell Res. 195,361-367 (1991). [CrossRef] [PubMed]
  3. Y. Takai, T. Masuko, and H. Takeuchi, "Lipid structure of cytotoxic granules in living human killer T lymphocytes studied by Raman microspectroscopy," Biochim. Biophys. Acta 1335,199-208 (1997). [CrossRef] [PubMed]
  4. C. Otto, N. M. Sijtsema, and J. Greve, "Confocal Raman microspectroscopy of the activation of single neutrophilic granulocytes," Eur. Biophys. J. 27,582-589 (1998). [CrossRef] [PubMed]
  5. Y.-S. Huang, T. Karashima, M. Yamamoto, and H. Hamaguchi, "Molecular-level pursuit of yeast mitosis by time- and space-resolved Raman spectroscopy," J. Raman Spectrosc. 34,1-3 (2003). [CrossRef]
  6. Y.-S. Huang, T. Karashima, M. Yamamoto, T. Ogura, and H. Hamaguchi, "Raman spectroscopic signature of life in a living yeast cell," J. Raman Spectrosc. 35,525-526 (2004). [CrossRef]
  7. Y.-S. Huang, T. Karashima, M. Yamamoto, and H. Hamaguchi, "Molecular-level investigation of the structure, transformation, and bioactivity of single living fission yeast cells by time- and space-resolved Raman Spectroscopy," Biochemistry 44,10009-10019 (2005). [CrossRef] [PubMed]
  8. Y. Naito, A. Toh-e,and H.-o. Hamaguchi, "In vivo time-resolved Raman imaging of a spontaneous death process of a single budding yeast cell," J. Raman Spectrosc. 36,837-839 (2005). [CrossRef]
  9. 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]
  10. 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]
  11. M. Hashimoto, T. Araki, and S. Kawata, "Molecular vibration imaging in the fingerprint region by use of coherent anti-Stokes Raman scattering microscopy with a collinear configuration," Opt. Lett. 25,1768-1770 (2000). [CrossRef]
  12. G. W. H. Wurpel, J. M. Schins, and M. Mueller, "Chemical specificity in three-dimensional imaging with multiplex coherent anti-Stokes Raman scattering microscopy," Opt. Lett. 27,1093-1095 (2002). [CrossRef]
  13. H. N. Paulsen, K. M. Hilligsoe, J. Thogersen, S. R. Keiding, and J. J. Larsen, "Coherent anti-Stokes Raman scattering microscopy with a photonic crystal fiber based light source," Opt. Lett. 28,1123-1125 (2003). [CrossRef] [PubMed]
  14. R. D. Schaller, J. Ziegelbauer, L. F. Lee, L. H. Haber, and R. J. Saykally, "Chemically selective imaging of subcellular structure in human hepatocytes with coherent anti-stokes Raman scattering (CARS) near-field scanning optical microscopy (NSOM)," J. Phys. Chem. B 106,8489-8492 (2002). [CrossRef]
  15. T. Ichimura, N. Hayazawa, M. Hashimoto, Y. Inouye, and S. Kawata, "Tip-enhanced coherent anti-stokes Raman scattering for vibrational nanoimaging," Phys. Rev. Lett. 92,220801 (2004). [CrossRef] [PubMed]
  16. C. Otto, A. Voroshilov, S. G. Kruglik, and J. Greve, "Vibrational bands of luminescent zinc(II)-octaethyl-porphyrin using a polarization-sensitive "microscopic" multiplex CARS technique," J. Raman Spectrosc. 32,495-501 (2001). [CrossRef]
  17. J.-X. 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]
  18. D. Oron, N. Dudovich, and Y. Silberberg, "Single-pulse phase-contrast nonlinear Raman Spectroscopy," Phys. Rev. Lett. 89,273001 (2002). [CrossRef]
  19. D. Oron, N. Dudovich, D. Yelin, and Y. Silberberg, "Narrow-band coherent anti-stokes Raman signals from broad-band pulses," Phys. Rev. Lett. 88,063004 (2002). [CrossRef] [PubMed]
  20. H. Kano, and H. Hamaguchi, "Dispersion-compensated supercontinuum generation for ultrabroadband multiplex coherent anti-Stokes Raman scattering spectroscopy," J. Raman Spectrosc., in press.
  21. S.-H. Lim, A. G. Caster, and S. R. Leone, "Single-pulse phase-control interferometric coherent anti-Stokes Raman scattering spectroscopy," Phys. Rev. A 30,2805-2807 (2005).
  22. I. G. Petrov, and V. V. Yakovlev, "Enhancing red-shifted white-light continuum generation in optical fibers for applications in nonlinear Raman microscopy," Opt. Express 13,1299 (2005). [CrossRef] [PubMed]
  23. http://www.opticsexpress.org/abstract.cfm?URI=OPEX-13-4-1299 [CrossRef]
  24. S. O. Konorov, D. A. Akimov, E. E. Serebryannikov, A. A. Ivanov, M. V. Alfimov, and A. M. Zheltikov, "Cross-correlation frequency-resolved optical gating coherent anti-Stokes Raman scattering with frequency-converting photonic-crystal fibers," Phys. Rev. E 70,057601 (2004). [CrossRef]
  25. H. Kano, and H. Hamaguchi, "Ultrabroadband (>2500 cm-1) Multiplex coherent anti-stokes raman scattering microspectroscopy using a supercontinuum generated from a photonic crystal fiber," Appl. Phys. Lett. 86,121113-121115 (2005). [CrossRef] [PubMed]
  26. H. Kano, and H. Hamaguchi, "Vibrationally resonant imaging of a single living cell by supercontinuum-based multiplex coherent anti-Stokes Raman scattering microspectroscopy," Opt. Express 13,1322-1327 (2005). http://www.opticsexpress.org/abstract.cfm?URI=OPEX-13-4-1322 [CrossRef] [PubMed]
  27. T. W. Kee, and M. T. Cicerone, "Simple approach to one-laser, broadband coherent anti-Stokes Raman scattering microscopy," Opt. Lett. 29,2701-2703 (2004). [CrossRef] [PubMed]
  28. C. McConnell, and E. Riis, "Photonic crystal fibre enables short-wavelength two-photon laser scanning fluorescence microscopy with fura-2," Phys. Med. Biol. 49,4757-4763 (2004). [CrossRef]
  29. K. Isobe, W. Watanabe, S. Matsunaga, T. Higashi, K. Fukui, and K. Itoh, "Multi-spectral two-photon excited fluorescence microscopy using supercontinuum light source," Jpn. J. Appl. Phys. Part 2 44,L167-L169 (2005). [CrossRef] [PubMed]
  30. J. A. Palero, V. O. Boer, J. C. Vijverberg, H. C. Gerritsen, and H. J. C. M. Sterenborg, "Short-wavelength two-photon excitation fluorescence microscopy of tryptophan with a photonic crystal fiber based light source," Opt. Express 13,5363-5368 (2005).
  31. http://www.opticsexpress.org/abstract.cfm?URI=OPEX-13-14-5363. [CrossRef] [PubMed]
  32. H. Kano, and H. Hamaguchi, "Vibrational imaging of a J-aggregate microcrystal using ultrabroadband multiplex coherent anti-Stokes Raman scattering microspectroscopy," submitted toVibrational Spectroscopy.
  33. H. Wang, Y. Fu, P. Zickmund, R. Shi, and J.-X. Cheng, "Coherent anti-Stokes Raman scattering imaging of live spinal tissues," Biophys. J. 89,581-591 (2005).

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