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Optics Express

Optics Express

  • Editor: Michael Duncan
  • Vol. 13, Iss. 4 — Feb. 21, 2005
  • pp: 1322–1327
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Vibrationally resonant imaging of a single living cell by supercontinuum-based multiplex coherent anti-Stokes Raman scattering microspectroscopy

Hideaki Kano and Hiro-o Hamaguchi  »View Author Affiliations


Optics Express, Vol. 13, Issue 4, pp. 1322-1327 (2005)
http://dx.doi.org/10.1364/OPEX.13.001322


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Abstract

Supercontinuum-based multiplex coherent anti-Stokes Raman scattering (CARS) microspectroscopy has been applied to vibrational imaging of a living fission yeast cell. We have successfully extracted only a vibrationally resonant CARS image from a characteristic spectral profile in the C-H stretching vibrational region. Using our simple but sensitive analysis, the vibrational contrast is significantly improved in comparison with a CARS imaging at a fixed Raman shift. The CARS image of a living yeast cell indicates several areas at which the signal is remarkably strong. They are considered to arise from mitochondria.

© 2005 Optical Society of America

1. Introduction

Coherent anti-Stokes Raman scattering (CARS) microscopy has become a powerful technique for three-dimensional vibrational imaging of chemical and biological samples [1

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]

8

8. H. Kano and H. Hamaguchi, “Near-infrared coherent anti-Stokes Raman scattering microscopy using supercontinuum generated from a photonic crystal fiber,” Appl. Phys. B B 80, 243–246 (2005). [CrossRef]

]. Recently, significant progress has been made of CARS microscopy such as near-infrared excitation [1

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]

], epi-detection [9

9. J.-X. Cheng, L. D. Book, and X. S. Xie, “Polarization coherent anti-Stokes Raman scattering microscopy,” Opt. Lett. 26, 1341–1343 (2001). [CrossRef]

], time-resolved detection [10

10. A. Volkmer, L. D. Book, and X. S. Xie, “Time-resolved coherent anti-Stokes Raman scattering microscopy: imaging based on Raman free induction decay,” Appl. Phys. Lett. 80, 1505–1507 (2002). [CrossRef]

], multiplex measurement [11

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

13

13. 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]

], and heterodyne detection [14

14. C. L. Evans, E. O. Potma, and X. S. Xie, “Coherent anti-Stokes Raman scattering spectral interferometry: determination of the real and imaginary components of nonlinear susceptibility chi(3) for vibrational microscopy,” Opt. Lett. 29, 2923–2925 (2004). [CrossRef]

]. Among them, multiplex CARS microspectrosopy is especially useful because it is capable of obtaining not only a CARS image but also CARS spectrum with a data-acquisition time faster than conventional spontaneous Raman microscopy. Recently, the spectral coverage of the multiplex CARS detection has been significantly broadened using a supercontinuum Stokes laser, which has been reported independently by Kee and Cicerone [15

15. 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]

] and by us [16

16. 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. (accepted). [PubMed]

]. In these setups, supercontinuum is generated from a tapered fiber [15

15. 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]

] or a photonic crystal fiber (PCF) [16

16. 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. (accepted). [PubMed]

] using a femtosecond Ti:Sapphire oscillator. Owing to a low threshold of the continuum generation [5

5. 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]

, 17

17. J. K. Ranka, R. S. Windeler, and A. J. Stentz, “Visible continuum generation in air-silica microstructure optical fibers with anomalous dispersion at 800 nm,” Opt. Lett. 25, 25–27 (2000). [CrossRef]

19

19. M. L. Hu, C. Y. Wang, Y. Li, Z. Wang, L. Chai, and A. M. Zheltikov “Multiplex frequency conversion of unamplified 30-fs Ti: sapphire laser pulses by an array of waveguiding wires in a random-hole microstructure fiber,” Opt. Exp. 12, 6129–6134 (2004), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-25-6129. [CrossRef]

], various kinds of applications such as spectroscopy [20

20. H. Kano and H. Hamaguchi, “Characterization of a supercontinuum generated from a photonic crystal fiber and its application to coherent Raman spectroscopy,” Opt. Lett. 28, 2360–2362 (2003). [CrossRef] [PubMed]

22

22. T. Nagahara, K. Imura, and H. Okamoto, “Time-resolved scanning near-field optical microscopy with supercontinuum light pulses generated in microstructure fiber,” Rev. Sci. Instrum. 75, 4528–4533 (2004). [CrossRef]

] and metrology [23

23. R. Holzwarth, T. Udem, T. W. Hänsch, J. C. Knight, W. J. Wadsworth, and P. S. J. Russell, “Optical frequency synthesizer for precision spectroscopy,” Phys. Rev. Lett. 85, 2264–2267 (2000). [CrossRef] [PubMed]

] have been reported only using a single Ti: Sapphire oscillator. It is also advantageous in microscopy because many studies on CARS microscopy require a complicated laser system such as an amplified laser source or two-synchronized oscillators. Using the supercontinuum as the ultrabroadband Stokes laser, ultrabroadband multiplex CARS microspectroscopy has been demonstrated over a 2500cm-1-wavenumber region, which spans from the fingerprint region to the C-H or O-H stretching region [15

15. 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]

, 16

16. 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. (accepted). [PubMed]

].

It is well known that the vibrationally resonant CARS signal is often overwhelmed by a nonresonant background, which results in degradation of the vibrational contrast of a CARS image [7

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

]. Several reports have been made of the suppression of the nonresonant background using the polarization-sensitive detection [9

9. J.-X. Cheng, L. D. Book, and X. S. Xie, “Polarization coherent anti-Stokes Raman scattering microscopy,” Opt. Lett. 26, 1341–1343 (2001). [CrossRef]

], epi-detection [24

24. A. Volkmer, J.-X. Cheng, and X. S. Xie, “Vibrational imaging with high sensitivity via epidetected coherent anti-Stokes Raman scattering microscopy,” Phys. Rev. Lett. 87, 023901–023904 (2001). [CrossRef]

], time-resolved detection [10

10. A. Volkmer, L. D. Book, and X. S. Xie, “Time-resolved coherent anti-Stokes Raman scattering microscopy: imaging based on Raman free induction decay,” Appl. Phys. Lett. 80, 1505–1507 (2002). [CrossRef]

], and heterodyne detection [14

14. C. L. Evans, E. O. Potma, and X. S. Xie, “Coherent anti-Stokes Raman scattering spectral interferometry: determination of the real and imaginary components of nonlinear susceptibility chi(3) for vibrational microscopy,” Opt. Lett. 29, 2923–2925 (2004). [CrossRef]

]. However, some techniques decrease the resonant CARS signal itself [9

9. J.-X. Cheng, L. D. Book, and X. S. Xie, “Polarization coherent anti-Stokes Raman scattering microscopy,” Opt. Lett. 26, 1341–1343 (2001). [CrossRef]

, 10

10. A. Volkmer, L. D. Book, and X. S. Xie, “Time-resolved coherent anti-Stokes Raman scattering microscopy: imaging based on Raman free induction decay,” Appl. Phys. Lett. 80, 1505–1507 (2002). [CrossRef]

], and the other requires complicated experimental setup for a local oscillator [14

14. C. L. Evans, E. O. Potma, and X. S. Xie, “Coherent anti-Stokes Raman scattering spectral interferometry: determination of the real and imaginary components of nonlinear susceptibility chi(3) for vibrational microscopy,” Opt. Lett. 29, 2923–2925 (2004). [CrossRef]

]. In the present study, we have applied ultrabroadband multiplex CARS microspectroscopy to a living yeast cell, and succeeded in obtaining a resonant CARS image with significantly enhanced vibrational contrast. Instead of employing complicated setup, we analyzed carefully the CARS spectral profile, and extracted only the resonant component. The key is an interference of the resonant CARS signal with the nonresonant background, which works as a local oscillator for the resonant CARS signal [25

25. G. W. H. Wurpel, J. M. Schins, and M. Müller, “Direct measurement of chain order in single phospholipid mono- and bilayers with multiplex CARS,” J. Phys. Chem. B 108, 3400–3403 (2004). [CrossRef]

].

2. Experimental

The ultrabroadband multiplex CARS microspectroscopy using a PCF is described elsewhere [16

16. 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. (accepted). [PubMed]

]. Briefly, an unamplified mode-locked Ti:Sapphire laser (Coherent, Vitesse-800) was used as a laser source. Typical duration, pulse energy, peak wavelength, and repetition rate were 100 fs, 12 nJ, 801 nm, and 80 MHz, respectively. A portion 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 (ω 1) and Stokes (ω 2) lasers, respectively. In order to obtain CARS spectrum with high frequency resolution, the pump laser pulses were spectrally filtered using a narrow-bandpass filter. The peak position and bandwidth were measured to be about 12490 cm-1 (800.6 nm) and 23 cm-1, respectively. For Raman resonance, the wavelength of the Stokes laser must be in the near-infrared (NIR) region because that of the pump laser is also in the NIR. The visible component in the supercontinuum was thereby blocked by a long-wavelength-pass filter. The pulse energy of the pump and Stokes lasers were 120 and 140 pJ, respectively. Two laser pulses were superimposed collinearly using an 800-nm Notch filter, and then tightly focused onto the sample with a 40×0.9 NA microscope objective. We used an inverted microscope (Nikon, TE2000-S), which was modified for our setup. The forward-propagating CARS signal was collected with a 40×0.6 NA microscope objective. After passing through 800-nm Notch and short-wavelength-pass filter, 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) with a step size of 153 nm in both the x and y directions. An exposure time for each point was 200 ms. The lateral and axial spatial resolutions in the present setup were 0.59±0.01µm and 4.2±0.3 µm, respectively. We used fission yeast Schizosaccharomyces pombe (S. pombe) as a sample [26

26. 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]

, 27

27. 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]

]. 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.

Fig. 1. Typical spectral profile of the CARS signal of a living yeast cell (red solid) and surrounding water (blue dotted)

3. Results and discussion

Typical spectral profiles of the CARS signal from a living yeast cell and surrounding water are shown in Fig. 1. The overall CARS spectral profile depends on the temporal overlap between the pump and Stokes laser pulses, because the supercontinuum Stokes pulses are temporally chirped due to a fiber dispersion [16

16. 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. (accepted). [PubMed]

]. By changing the delay time between the pump and Stokes laser pulses, we can control the spectral coverage of the CARS measurement and a wavenumber region at which the CARS signal is obtained with high detection efficiency. In the present setup, the delay time is optimized for efficient detection of the CARS signal due to the C-H stretching vibrational mode. As clearly shown in Fig. 1, a dispersive lineshape is observed around 2856 cm-1, which is due to an interference of the resonant C-H stretching CARS signal with the nonresonant background. Because of this interference, the resonant CARS signal is intrinsically heterodyne-detected by the nonresonant background. On the other hand, the vibrationally nonresonant background signal of water shows a featureless spectral profile that represents that of the supercontinuum. Figure 2(a) and (b) shows the CARS images of living yeast cells at the Raman shift of 2856 and 2200 cm-1, respectively. It is noted that both images are simultaneously obtained by the multiplex CARS detection. The CARS spectra of a yeast cell (red solid) and water background (blue dotted) in Fig. 1 are obtained at (x, y)=(-1.98 µm, -1.37 µm) and (-3.05 µm, 3.05 µm) positions, which are indicated as black and white crosses in Fig. 2(a), respectively. At 2856 cm-1, the resonant CARS signal is observed mainly from yeast cells, although the vibrational contrast is degraded due to the nonresonant background surrounding the cells. The dark and round region around the center of the yeast cell is most probably due to a nucleus. On the other hand, the contrast is dramatically decreased in Fig. 2(b) because there is no vibrational resonance at this Raman shift. Both in Fig 2(a) and (b), an outline of a yeast cell is observed as a dark region. The observed pattern is caused by a distortion of the focus spots due to refractive index mismatch [7

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

].

Fig. 2. CARS lateral images of living yeast cells at the Raman shift of 2856 cm-1 (a) and 2200 cm-1 (b). The black and white crosses in (a) correspond to the positions at which the spectral profiles are shown in Fig. 1.

Fig. 3. Vibrationally resonant CARS imaging using the differentiation method (a) and a method using a conventional lineshape function described in Eq. (1) (b). Note that the contrast and signal-to-noise ratio is dramatically improved in comparison with Fig. 2(a).

The spectral profile of the CARS signal is often fitted by the following function [4

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

, 11

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

, 13

13. 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]

, 25

25. G. W. H. Wurpel, J. M. Schins, and M. Müller, “Direct measurement of chain order in single phospholipid mono- and bilayers with multiplex CARS,” J. Phys. Chem. B 108, 3400–3403 (2004). [CrossRef]

];

I(ω)=ANReiϕ+RARΓRΓRi(ωΩR)2.
(1)

Here I(ω) is the observed signal intensity, A NR and ϕ the amplitude and phase of the nonresonant background, A R the amplitude of the Raman resonance signal, ΩR the Raman frequency of the molecule, and ΓR the dephasing rate of the Raman band. It is informative to compare our method with the conventional analysis using Eq. (1). The spectral profile of the CARS signal in Fig. 1 is fitted by Eq. (1) in wavenumber region from 2588 to 3135 cm-1. Here we assumed that the amplitude of the nonresonant background is described by a second-order polynomial function, and that only one Raman resonance is observed. The Raman frequency, dephasing rate, and phase of the nonresonant background, namely, ΩR, ΓR, and ϕ, are determined to be 2894 cm-1, 69 cm-1, and 1.8 radian, respectively. Since there are many fitting parameters, the values ΩR, ΓR, and ϕ are fixed in the fitting procedure for each spatial point. The resultant CARS image using the amplitude A R in Eq. (1) is shown in Fig. 3(b). The original data is the same as that of Fig. 2. The CARS image in Fig. 3(b) is very similar to Fig. 3(a). It means that the vibrationally resonant CARS image can be extracted not only using the complicated lineshape function described in Eq. (1) but also using the simple differentiation technique demonstrated in the present paper.

4. Conclusion

In conclusion, vibrational imaging of a living yeast cell has been performed by supercontinuum-based multiplex CARS microspectroscopy. Owing to the broadband multiplex CARS detection, the spectral profile of the CARS signal can be analyzed in detail. The differentiation method enables us to extract only the resonant CARS image with the high vibrational contrast. The clear vibrational image of a yeast cell itself and mitochondria is successfully obtained. In comparison with the conventional fitting analysis, the differentiation technique is simple and sensitive, and is extremely useful in enhancing the vibrational contrast.

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 Ms. 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.

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]

2.

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]

3.

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]

4.

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

5.

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]

6.

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, 220801–220804 (2004). [CrossRef] [PubMed]

7.

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

8.

H. Kano and H. Hamaguchi, “Near-infrared coherent anti-Stokes Raman scattering microscopy using supercontinuum generated from a photonic crystal fiber,” Appl. Phys. B B 80, 243–246 (2005). [CrossRef]

9.

J.-X. Cheng, L. D. Book, and X. S. Xie, “Polarization coherent anti-Stokes Raman scattering microscopy,” Opt. Lett. 26, 1341–1343 (2001). [CrossRef]

10.

A. Volkmer, L. D. Book, and X. S. Xie, “Time-resolved coherent anti-Stokes Raman scattering microscopy: imaging based on Raman free induction decay,” Appl. Phys. Lett. 80, 1505–1507 (2002). [CrossRef]

11.

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

12.

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

13.

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]

14.

C. L. Evans, E. O. Potma, and X. S. Xie, “Coherent anti-Stokes Raman scattering spectral interferometry: determination of the real and imaginary components of nonlinear susceptibility chi(3) for vibrational microscopy,” Opt. Lett. 29, 2923–2925 (2004). [CrossRef]

15.

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]

16.

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. (accepted). [PubMed]

17.

J. K. Ranka, R. S. Windeler, and A. J. Stentz, “Visible continuum generation in air-silica microstructure optical fibers with anomalous dispersion at 800 nm,” Opt. Lett. 25, 25–27 (2000). [CrossRef]

18.

M. L. Hu, C. Y. Wang, L. Chai, and A. M. Zheltikov “Frequency-tunable anti-Stokes line emission by eigenmodes of a birefringent microstructure fiber,” Opt. Exp. 12, 1932–1937 (2004), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-9-1932. [CrossRef]

19.

M. L. Hu, C. Y. Wang, Y. Li, Z. Wang, L. Chai, and A. M. Zheltikov “Multiplex frequency conversion of unamplified 30-fs Ti: sapphire laser pulses by an array of waveguiding wires in a random-hole microstructure fiber,” Opt. Exp. 12, 6129–6134 (2004), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-25-6129. [CrossRef]

20.

H. Kano and H. Hamaguchi, “Characterization of a supercontinuum generated from a photonic crystal fiber and its application to coherent Raman spectroscopy,” Opt. Lett. 28, 2360–2362 (2003). [CrossRef] [PubMed]

21.

H. Kano and H. Hamaguchi, “Femtosecond coherent anti-Stokes Raman scattering spectroscopy using a supercontinuum generated from a photonic crystal fiber,” Appl. Phys. Lett. 85, 4298–4300 (2004). [CrossRef]

22.

T. Nagahara, K. Imura, and H. Okamoto, “Time-resolved scanning near-field optical microscopy with supercontinuum light pulses generated in microstructure fiber,” Rev. Sci. Instrum. 75, 4528–4533 (2004). [CrossRef]

23.

R. Holzwarth, T. Udem, T. W. Hänsch, J. C. Knight, W. J. Wadsworth, and P. S. J. Russell, “Optical frequency synthesizer for precision spectroscopy,” Phys. Rev. Lett. 85, 2264–2267 (2000). [CrossRef] [PubMed]

24.

A. Volkmer, J.-X. Cheng, and X. S. Xie, “Vibrational imaging with high sensitivity via epidetected coherent anti-Stokes Raman scattering microscopy,” Phys. Rev. Lett. 87, 023901–023904 (2001). [CrossRef]

25.

G. W. H. Wurpel, J. M. Schins, and M. Müller, “Direct measurement of chain order in single phospholipid mono- and bilayers with multiplex CARS,” J. Phys. Chem. B 108, 3400–3403 (2004). [CrossRef]

26.

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]

27.

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]

OCIS Codes
(110.0180) Imaging systems : Microscopy
(300.6230) Spectroscopy : Spectroscopy, coherent anti-Stokes Raman scattering

ToC Category:
Research Papers

History
Original Manuscript: February 8, 2005
Revised Manuscript: February 15, 2005
Published: February 21, 2005

Citation
Hideaki Kano and Hiro-o 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.opticsinfobase.org/oe/abstract.cfm?URI=oe-13-4-1322


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References

  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]
  2. 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]
  3. 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]
  4. M. Müller and J. M. Schins, "Imaging the thermodynamic state of lipid membranes with multiplex CARS microscopy," J. Phys. Chem. B 106, 3715-3723 (2002). [CrossRef]
  5. 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]
  6. 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, 220801-220804 (2004). [CrossRef] [PubMed]
  7. J.-X. Cheng and X. S. Xie, "Coherent anti-Stokes Raman scattering microscopy: instrumentation, theory, and applications," J. Phys. Chem. B 108, 827-840 (2004). [CrossRef]
  8. H. Kano and H. Hamaguchi, "Near-infrared coherent anti-Stokes Raman scattering microscopy using supercontinuum generated from a photonic crystal fiber," Appl. Phys. B B80, 243-246 (2005). [CrossRef]
  9. J.-X. Cheng, L. D. Book, and X. S. Xie, "Polarization coherent anti-Stokes Raman scattering microscopy," Opt. Lett. 26, 1341-1343 (2001). [CrossRef]
  10. A. Volkmer, L. D. Book, and X. S. Xie, "Time-resolved coherent anti-Stokes Raman scattering microscopy: imaging based on Raman free induction decay," Appl. Phys. Lett. 80, 1505-1507 (2002). [CrossRef]
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