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]. 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].The “Raman spectroscopic signature of life” has been found in living fission [5–7] and budding [8] 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]. In particular, multiplex CARS microspectroscopy is promising because of its capability to obtain vibrational spectra efficiently [12, 16–19]. 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, 21]. 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, 16–19], the spectral coverage has been significantly broadened using the supercontinuum light source generated from a photonic crystal fiber [22–25] or a tapered fiber [26]. Recently, the spectral coverage of the multiplex CARS microspectroscopy has been extended to be more than 2800 cm-1 [20], 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]. 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]. 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]. We used fission yeast Schizosaccharomyces pombe (S. pombe) as a sample [5–7]. 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

Figure 1(a) shows typical spectral profiles of the CARS signal of a living yeast cell (red) and surrounding water (blue). As clearly shown, a strong signal is observed inside of the yeast cell at the Raman shift of 2840 cm-1. This band originates from CH2 stretching vibrational modes, which shows a slightly dispersive lineshape due to an interference with the nonresonant background. In a previous study, we have extracted only the vibrationally resonant signal using a differentiation method [25]. In the present study, however, the vibrational signal is dominantly observed probably due to the improved spatial resolution and spatial overlap between the pump and Stokes laser pulses. On the basis of our previous spontaneous Raman [5–7] and CARS [25] studies, the signal at the Raman shift of 2840 cm-1 is found especially in mitochondria, because mitochondrion is an organelle containing a high concentration of phospholipids. Figure 1(b) shows a CARS image of living cells at the Raman shift of 2840 cm-1. The CARS spectra of a yeast cell (red) and water background (blue) in Fig. 1(a) are obtained at (x, y)=(5.65 μm, -3.05 μm) and (6.71 μm, 1.22 μm), which are indicated as black and white crosses in Fig. 1(b), respectively. Figure 1(b) indicates various yeast cells at different stages of the cell division cycle. Especially, a septum is visualized in the yeast cell around the center of Fig. 1(b). The septum is composed of carbohydrates such as saccharide, which is also rich in CH2 bonds. The CARS signal at the edge of the yeast cell decreases mainly due to the imperfect focusing caused by the refractive index mismatch between the yeast cell and surrounding water.

 

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.

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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.

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Next we shall focus on the two-photon excitation fluorescence (TPEF). As described in the previous section, the nuclei of the yeast cells in the present study are tagged by GFP. Figure 3(a) shows the CARS and the TPEF spectra obtained in 100 ms exposure time. The weak but distinguishable peak is observed around 506 nm. Taking into account of the spectral profile of this signal, it is assigned to the TPEF signal due to GFP. Owing to the high quantum yield of GFP, the fluorescence intensity is enough to be detected with such short data-acquisition time. Figures 3(b) and 3(d) show CARS and TPEF images at zero delay time between the pump and Stokes pulses. It should be emphasized that both images of multiplex CARS and TPEF signals are obtained simultaneously. Since there is no overlap between the CARS and TPEF signals in spectral domain, it can be easily differentiated using the spectrometer. Although dual imaging of the CARS and the TPEF signals have been reported using two synchronized Ti:Sapphire oscillators [31], full spectral information is obtained for the first time in the present study. The CARS and TPEF signals indicated as red and green curves in Fig. 3(a) are obtained at (x, y)=(0.31 μm, -2.29 μm) and (4.27 μm, 1.53 μm), respectively. A yeast cell around the center of Figs. 3(b) and (d) shows two spots due to GFP in Fig. 3(d). It means that the yeast cell is in the M phase. Compared with Fig 1(b), septum is not clearly visualized by CH2 stretching vibrational mode. From our previous study [5], Raman spectrum of a matured septum is different from that of a primary septum. From this result, the CARS signal intensity due to CH2 stretching vibrational mode can be used for monitoring the evolution of the septum. Figures 3(c) and 3(e) show CARS and TPEF images at -4-ps delay time between the pump and Stokes pulses. Since the CARS process occurs through a virtual electronic transition, no signal is observed when there is no temporal overlap between the pump and Stokes pulses at such a large delay. On the other hand, TPEF signal is still observed even without temporal overlap. It should be noted, however, that the signal intensity is drastically decreased by a factor of 3.4. The signal is, therefore, ascribed to degenerate 2ω1- and/or 2ω2-photon processes. In the present experimental condition, the TPEF signal due to the 2ω2-photon process was negligible. In order to evaluate the signal intensity only due to the non-degenerated (ω12)-photon process, we have subtracted the TPEF signal intensity in Fig. 3(e) from that in Fig. 3(d). The result is depicted in Fig. 3(f). It is clear that the TPEF signal due to the (ω12)-photon process is larger than that due to the 2ω1-photon processes. It can be explained by the following two reasons. First, the TPEF signal intensity in Fig. 3(f) is proportional to the intensity both of the ω1 and the ω2 pulses. On the other hand, the TPEF signal intensity in Fig. 3(e) depends quadratically on the intensity of the ω1 pulses. Therefore, the threshold of the laser power in the former is lower than that in the latter. Second, the wide spectral bandwidth of the Stokes pulse works as an efficient twophoton excitation light source. On the other hand, the degenerate two-photon process provides a bandwidth much narrower than that of the non-degenerated (ω12)-photon process. Furthermore, a spectrally filtered or a temporally chirped supercontiuum with a narrow-band pump pulse can be used for an ideal light source to obtain fluorescence excitation spectrum of a two-photon allowed electronic state.

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.

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Fig. 4. In-vivo multi-nonlinear optical imaging of yeast cells. The CARS and TPEF images are indicated as red and green, respectively.

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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]  

References

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  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]
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    [CrossRef]
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    [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]
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    [CrossRef] [PubMed]
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    [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).

2005 (9)

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]

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]

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]

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]

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).

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]

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]

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).

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).

2004 (5)

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]

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]

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]

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]

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]

2003 (2)

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]

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]

2002 (6)

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]

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]

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]

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

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]

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]

2001 (1)

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]

2000 (1)

1999 (1)

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]

1998 (1)

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]

1997 (1)

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]

1991 (1)

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]

1990 (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]

Akimov, D. A.

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]

Alfimov, M. V.

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]

Araki, T.

Arndt-Jovin, D. J.

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]

Boer, V. O.

Book, L. D.

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]

Caster, A. G.

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).

Cheng, J.-X.

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).

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]

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]

Cicerone, M. T.

de Mul, F. F.

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]

De Mul, F. F. M.

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]

Dudovich, N.

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

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]

Fu, Y.

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).

Fukui, K.

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]

Gerritsen, H. C.

Greve, J.

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]

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]

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]

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]

Haber, L. H.

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]

Hamaguchi, H.

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]

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]

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]

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]

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]

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

Hamaguchi, H.-o.

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]

Hashimoto, M.

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]

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]

Hayazawa, N.

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]

Higashi, T.

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]

Hilligsoe, K. M.

Holtom, G. R.

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]

Huang, Y.-S.

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]

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]

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]

Ichimura, T.

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]

Inouye, Y.

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]

Isobe, K.

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]

Itoh, K.

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]

Ivanov, A. A.

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]

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]

Jovin, T. M.

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]

Kano, H.

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]

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]

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

Karashima, T.

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]

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]

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]

Kawata, S.

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]

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]

Kee, T. W.

Keiding, S. R.

Konorov, S. O.

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]

Kruglik, S. G.

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]

Larsen, J. J.

Lee, L. F.

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]

Leone, S. R.

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).

Lim, S.-H.

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).

Masuko, T.

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]

Matsunaga, S.

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]

McConnell, C.

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]

Mueller, M.

Naito, Y.

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]

Ogura, T.

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]

Olminkhof, J. H.

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]

Oron, D.

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

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]

Otto, C.

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]

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]

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]

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]

Palero, J. A.

Paulsen, H. N.

Petrov, I. G.

Puppels, G. J.

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]

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]

Riis, E.

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]

Robert-Nicoud, M.

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]

Saykally, R. J.

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]

Schaller, R. D.

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]

Schins, J. M.

Segers-Nolten, G. M.

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]

Serebryannikov, E. E.

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]

Shi, R.

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).

Sijtsema, N. M.

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]

Silberberg, Y.

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]

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

Sterenborg, H. J. C. M.

Takai, Y.

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]

Takeuchi, H.

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]

Thogersen, J.

Toh-e, A.

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]

Vijverberg, J. C.

Volkmer, A.

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]

Voroshilov, A.

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]

Wang, H.

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).

Watanabe, W.

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]

Wurpel, G. W. H.

Xie, X. S.

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]

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]

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]

Yakovlev, V. V.

Yamamoto, M.

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]

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]

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]

Yelin, D.

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]

Zheltikov, A. M.

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]

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).
[CrossRef] [PubMed]

Zickmund, P.

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).

Ziegelbauer, J.

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]

Zumbusch, A.

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]

Appl. Phys. Lett. (1)

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]

Biochemistry (1)

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]

Biochim. Biophys. Acta (1)

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]

Biophys. J. (2)

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]

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).

Eur. Biophys. J. (1)

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]

Exp. Cell Res. (1)

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]

J. Phys. Chem. B (2)

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]

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]

J. Raman Spectrosc. (5)

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]

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]

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]

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]

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

Jpn. J. Appl. Phys (1)

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]

Nature (London, United Kingdom) (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]

Opt. Express (3)

Opt. Lett. (4)

Phys. Med. Biol. (1)

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]

Phys. Rev. A (1)

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).

Phys. Rev. E (1)

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]

Phys. Rev. Lett. (4)

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

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]

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]

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]

Other (3)

http://www.opticsexpress.org/abstract.cfm?URI=OPEX-13-14-5363.
[CrossRef] [PubMed]

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

http://www.opticsexpress.org/abstract.cfm?URI=OPEX-13-4-1299
[CrossRef]

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Figures (4)

Fig. 1.
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.

Fig. 2.
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.

Fig. 3.
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.
Fig. 4.

In-vivo multi-nonlinear optical imaging of yeast cells. The CARS and TPEF images are indicated as red and green, respectively.

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