Abstract

We use the time-lens concept to demonstrate a new scheme for synchronization of two pulsed light sources for biological imaging. An all fiber, 1064 nm time-lens source is synchronized to a picosecond solid-state Ti: Sapphire mode-locked laser by using the mode-locked laser pulses as the clock. We demonstrate the application of this synchronized source for CARS and SRS imaging by imaging mouse tissues. Synchronized two wavelength pulsed source is an important technical difficulty for CARS and SRS imaging. The time-lens source demonstrated here may provide an all fiber, user friendly alternative for future SRS imaging.

©2010 Optical Society of America

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References

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  1. A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett. 82(20), 4142 (1999).
    [Crossref]
  2. C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science 322(5909), 1857–1861 (2008).
    [Crossref] [PubMed]
  3. C. L. Evans, E. O. Potma, M. Puoris’haag, D. Côté, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. U.S.A. 102(46), 16807–16812 (2005).
    [Crossref] [PubMed]
  4. C. L. Evans, X. Xu, S. Kesari, X. S. Xie, S. T. C. Wong, and G. S. Young, “Chemically-selective imaging of brain structures with CARS microscopy,” Opt. Express 15(19), 12076–12087 (2007).
    [Crossref] [PubMed]
  5. T. Baldacchini and R. Zadoyan, “In situ and real time monitoring of two-photon polymerization using broadband coherent anti-Stokes Raman scattering microscopy,” Opt. Express 18(18), 19219–19231 (2010).
    [Crossref] [PubMed]
  6. F. Svedberg, C. Brackmann, T. Hellerer, and A. J. Enejder, “Nonlinear microscopy with fiber laser continuum excitation,” J. Biomed. Opt. 15(2), 026026 (2010).
    [Crossref] [PubMed]
  7. A. F. Pegoraro, A. Ridsdale, D. J. Moffatt, J. P. Pezacki, B. K. Thomas, L. Fu, L. Dong, M. E. Fermann, and A. Stolow, “All-fiber CARS microscopy of live cells,” Opt. Express 17(23), 20700–20706 (2009).
    [Crossref] [PubMed]
  8. G. Krauss, T. Hanke, A. Sell, D. Träutlein, A. Leitenstorfer, R. Selm, M. Winterhalder, and A. Zumbusch, “Compact coherent anti-Stokes Raman scattering microscope based on a picosecond two-color Er:fiber laser system,” Opt. Lett. 34(18), 2847–2849 (2009).
    [Crossref] [PubMed]
  9. B. H. Kolner, “Active pulse-compression using an integrated electro-optic phase modulator,” Appl. Phys. Lett. 52(14), 1122–1124 (1988).
    [Crossref]
  10. D. H. Broaddus, M. A. Foster, O. Kuzucu, A. C. Turner-Foster, K. W. Koch, M. Lipson, and A. L. Gaeta, “Temporal-imaging system with simple external-clock triggering,” Opt. Express 18(13), 14262–14269 (2010).
    [Crossref] [PubMed]
  11. Z. Jiang, D. E. Leaird, and A. M. Weiner, “Optical processing based on spectral line-by-line pulse shaping on a phase-modulated CW laser,” IEEE J. Quantum Electron. 42(7), 657–665 (2006).
    [Crossref]
  12. J. van Howe, J. Hansryd, and C. Xu, “Multiwavelength pulse generator using time-lens compression,” Opt. Lett. 29(13), 1470–1472 (2004).
    [Crossref] [PubMed]
  13. J. van Howe, J. H. Lee, and C. Xu, “Generation of 3.5 nJ femtosecond pulses from a continuous-wave laser without mode locking,” Opt. Lett. 32(11), 1408–1410 (2007).
    [Crossref] [PubMed]
  14. E. O. Potma, D. J. Jones, J. X. Cheng, X. S. Xie, and J. Ye, “High-sensitivity coherent anti-Stokes Raman scattering microscopy with two tightly synchronized picosecond lasers,” Opt. Lett. 27(13), 1168–1170 (2002).
    [Crossref]
  15. W. Min, S. Lu, S. Chong, R. Roy, G. R. Holtom, and X. S. Xie, “Imaging chromophores with undetectable fluorescence by stimulated emission microscopy,” Nature 461(7267), 1105–1109 (2009).
    [Crossref] [PubMed]
  16. D. Fu, T. Ye, T. E. Matthews, B. J. Chen, G. Yurtserver, and W. S. Warren, “High-resolution in vivo imaging of blood vessels without labeling,” Opt. Lett. 32(18), 2641–2643 (2007).
    [Crossref] [PubMed]
  17. D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, “Two-color, two-photon, and excited-state absorption microscopy,” J. Biomed. Opt. 12(5), 054004 (2007).
    [Crossref] [PubMed]

2010 (3)

2009 (3)

2008 (1)

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science 322(5909), 1857–1861 (2008).
[Crossref] [PubMed]

2007 (4)

2006 (1)

Z. Jiang, D. E. Leaird, and A. M. Weiner, “Optical processing based on spectral line-by-line pulse shaping on a phase-modulated CW laser,” IEEE J. Quantum Electron. 42(7), 657–665 (2006).
[Crossref]

2005 (1)

C. L. Evans, E. O. Potma, M. Puoris’haag, D. Côté, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. U.S.A. 102(46), 16807–16812 (2005).
[Crossref] [PubMed]

2004 (1)

2002 (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(20), 4142 (1999).
[Crossref]

1988 (1)

B. H. Kolner, “Active pulse-compression using an integrated electro-optic phase modulator,” Appl. Phys. Lett. 52(14), 1122–1124 (1988).
[Crossref]

Baldacchini, T.

Brackmann, C.

F. Svedberg, C. Brackmann, T. Hellerer, and A. J. Enejder, “Nonlinear microscopy with fiber laser continuum excitation,” J. Biomed. Opt. 15(2), 026026 (2010).
[Crossref] [PubMed]

Broaddus, D. H.

Chen, B. J.

Cheng, J. X.

Chong, S.

W. Min, S. Lu, S. Chong, R. Roy, G. R. Holtom, and X. S. Xie, “Imaging chromophores with undetectable fluorescence by stimulated emission microscopy,” Nature 461(7267), 1105–1109 (2009).
[Crossref] [PubMed]

Côté, D.

C. L. Evans, E. O. Potma, M. Puoris’haag, D. Côté, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. U.S.A. 102(46), 16807–16812 (2005).
[Crossref] [PubMed]

Dong, L.

Enejder, A. J.

F. Svedberg, C. Brackmann, T. Hellerer, and A. J. Enejder, “Nonlinear microscopy with fiber laser continuum excitation,” J. Biomed. Opt. 15(2), 026026 (2010).
[Crossref] [PubMed]

Evans, C. L.

C. L. Evans, X. Xu, S. Kesari, X. S. Xie, S. T. C. Wong, and G. S. Young, “Chemically-selective imaging of brain structures with CARS microscopy,” Opt. Express 15(19), 12076–12087 (2007).
[Crossref] [PubMed]

C. L. Evans, E. O. Potma, M. Puoris’haag, D. Côté, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. U.S.A. 102(46), 16807–16812 (2005).
[Crossref] [PubMed]

Fermann, M. E.

Foster, M. A.

Freudiger, C. W.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science 322(5909), 1857–1861 (2008).
[Crossref] [PubMed]

Fu, D.

D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, “Two-color, two-photon, and excited-state absorption microscopy,” J. Biomed. Opt. 12(5), 054004 (2007).
[Crossref] [PubMed]

D. Fu, T. Ye, T. E. Matthews, B. J. Chen, G. Yurtserver, and W. S. Warren, “High-resolution in vivo imaging of blood vessels without labeling,” Opt. Lett. 32(18), 2641–2643 (2007).
[Crossref] [PubMed]

Fu, L.

Gaeta, A. L.

Hanke, T.

Hansryd, J.

He, C.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science 322(5909), 1857–1861 (2008).
[Crossref] [PubMed]

Hellerer, T.

F. Svedberg, C. Brackmann, T. Hellerer, and A. J. Enejder, “Nonlinear microscopy with fiber laser continuum excitation,” J. Biomed. Opt. 15(2), 026026 (2010).
[Crossref] [PubMed]

Holtom, G. R.

W. Min, S. Lu, S. Chong, R. Roy, G. R. Holtom, and X. S. Xie, “Imaging chromophores with undetectable fluorescence by stimulated emission microscopy,” Nature 461(7267), 1105–1109 (2009).
[Crossref] [PubMed]

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science 322(5909), 1857–1861 (2008).
[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(20), 4142 (1999).
[Crossref]

Jiang, Z.

Z. Jiang, D. E. Leaird, and A. M. Weiner, “Optical processing based on spectral line-by-line pulse shaping on a phase-modulated CW laser,” IEEE J. Quantum Electron. 42(7), 657–665 (2006).
[Crossref]

Jones, D. J.

Kang, J. X.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science 322(5909), 1857–1861 (2008).
[Crossref] [PubMed]

Kesari, S.

Koch, K. W.

Kolner, B. H.

B. H. Kolner, “Active pulse-compression using an integrated electro-optic phase modulator,” Appl. Phys. Lett. 52(14), 1122–1124 (1988).
[Crossref]

Krauss, G.

Kuzucu, O.

Leaird, D. E.

Z. Jiang, D. E. Leaird, and A. M. Weiner, “Optical processing based on spectral line-by-line pulse shaping on a phase-modulated CW laser,” IEEE J. Quantum Electron. 42(7), 657–665 (2006).
[Crossref]

Lee, J. H.

Leitenstorfer, A.

Lin, C. P.

C. L. Evans, E. O. Potma, M. Puoris’haag, D. Côté, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. U.S.A. 102(46), 16807–16812 (2005).
[Crossref] [PubMed]

Lipson, M.

Lu, S.

W. Min, S. Lu, S. Chong, R. Roy, G. R. Holtom, and X. S. Xie, “Imaging chromophores with undetectable fluorescence by stimulated emission microscopy,” Nature 461(7267), 1105–1109 (2009).
[Crossref] [PubMed]

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science 322(5909), 1857–1861 (2008).
[Crossref] [PubMed]

Matthews, T. E.

D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, “Two-color, two-photon, and excited-state absorption microscopy,” J. Biomed. Opt. 12(5), 054004 (2007).
[Crossref] [PubMed]

D. Fu, T. Ye, T. E. Matthews, B. J. Chen, G. Yurtserver, and W. S. Warren, “High-resolution in vivo imaging of blood vessels without labeling,” Opt. Lett. 32(18), 2641–2643 (2007).
[Crossref] [PubMed]

Min, W.

W. Min, S. Lu, S. Chong, R. Roy, G. R. Holtom, and X. S. Xie, “Imaging chromophores with undetectable fluorescence by stimulated emission microscopy,” Nature 461(7267), 1105–1109 (2009).
[Crossref] [PubMed]

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science 322(5909), 1857–1861 (2008).
[Crossref] [PubMed]

Moffatt, D. J.

Pegoraro, A. F.

Pezacki, J. P.

Potma, E. O.

C. L. Evans, E. O. Potma, M. Puoris’haag, D. Côté, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. U.S.A. 102(46), 16807–16812 (2005).
[Crossref] [PubMed]

E. O. Potma, D. J. Jones, J. X. Cheng, X. S. Xie, and J. Ye, “High-sensitivity coherent anti-Stokes Raman scattering microscopy with two tightly synchronized picosecond lasers,” Opt. Lett. 27(13), 1168–1170 (2002).
[Crossref]

Puoris’haag, M.

C. L. Evans, E. O. Potma, M. Puoris’haag, D. Côté, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. U.S.A. 102(46), 16807–16812 (2005).
[Crossref] [PubMed]

Ridsdale, A.

Roy, R.

W. Min, S. Lu, S. Chong, R. Roy, G. R. Holtom, and X. S. Xie, “Imaging chromophores with undetectable fluorescence by stimulated emission microscopy,” Nature 461(7267), 1105–1109 (2009).
[Crossref] [PubMed]

Saar, B. G.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science 322(5909), 1857–1861 (2008).
[Crossref] [PubMed]

Sell, A.

Selm, R.

Stolow, A.

Svedberg, F.

F. Svedberg, C. Brackmann, T. Hellerer, and A. J. Enejder, “Nonlinear microscopy with fiber laser continuum excitation,” J. Biomed. Opt. 15(2), 026026 (2010).
[Crossref] [PubMed]

Thomas, B. K.

Träutlein, D.

Tsai, J. C.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science 322(5909), 1857–1861 (2008).
[Crossref] [PubMed]

Turner-Foster, A. C.

van Howe, J.

Warren, W. S.

D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, “Two-color, two-photon, and excited-state absorption microscopy,” J. Biomed. Opt. 12(5), 054004 (2007).
[Crossref] [PubMed]

D. Fu, T. Ye, T. E. Matthews, B. J. Chen, G. Yurtserver, and W. S. Warren, “High-resolution in vivo imaging of blood vessels without labeling,” Opt. Lett. 32(18), 2641–2643 (2007).
[Crossref] [PubMed]

Weiner, A. M.

Z. Jiang, D. E. Leaird, and A. M. Weiner, “Optical processing based on spectral line-by-line pulse shaping on a phase-modulated CW laser,” IEEE J. Quantum Electron. 42(7), 657–665 (2006).
[Crossref]

Winterhalder, M.

Wong, S. T. C.

Xie, X. S.

W. Min, S. Lu, S. Chong, R. Roy, G. R. Holtom, and X. S. Xie, “Imaging chromophores with undetectable fluorescence by stimulated emission microscopy,” Nature 461(7267), 1105–1109 (2009).
[Crossref] [PubMed]

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science 322(5909), 1857–1861 (2008).
[Crossref] [PubMed]

C. L. Evans, X. Xu, S. Kesari, X. S. Xie, S. T. C. Wong, and G. S. Young, “Chemically-selective imaging of brain structures with CARS microscopy,” Opt. Express 15(19), 12076–12087 (2007).
[Crossref] [PubMed]

C. L. Evans, E. O. Potma, M. Puoris’haag, D. Côté, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. U.S.A. 102(46), 16807–16812 (2005).
[Crossref] [PubMed]

E. O. Potma, D. J. Jones, J. X. Cheng, X. S. Xie, and J. Ye, “High-sensitivity coherent anti-Stokes Raman scattering microscopy with two tightly synchronized picosecond lasers,” Opt. Lett. 27(13), 1168–1170 (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(20), 4142 (1999).
[Crossref]

Xu, C.

Xu, X.

Ye, J.

Ye, T.

D. Fu, T. Ye, T. E. Matthews, B. J. Chen, G. Yurtserver, and W. S. Warren, “High-resolution in vivo imaging of blood vessels without labeling,” Opt. Lett. 32(18), 2641–2643 (2007).
[Crossref] [PubMed]

D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, “Two-color, two-photon, and excited-state absorption microscopy,” J. Biomed. Opt. 12(5), 054004 (2007).
[Crossref] [PubMed]

Young, G. S.

Yurtserver, G.

Yurtsever, G.

D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, “Two-color, two-photon, and excited-state absorption microscopy,” J. Biomed. Opt. 12(5), 054004 (2007).
[Crossref] [PubMed]

Zadoyan, R.

Zumbusch, A.

Appl. Phys. Lett. (1)

B. H. Kolner, “Active pulse-compression using an integrated electro-optic phase modulator,” Appl. Phys. Lett. 52(14), 1122–1124 (1988).
[Crossref]

IEEE J. Quantum Electron. (1)

Z. Jiang, D. E. Leaird, and A. M. Weiner, “Optical processing based on spectral line-by-line pulse shaping on a phase-modulated CW laser,” IEEE J. Quantum Electron. 42(7), 657–665 (2006).
[Crossref]

J. Biomed. Opt. (2)

D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, “Two-color, two-photon, and excited-state absorption microscopy,” J. Biomed. Opt. 12(5), 054004 (2007).
[Crossref] [PubMed]

F. Svedberg, C. Brackmann, T. Hellerer, and A. J. Enejder, “Nonlinear microscopy with fiber laser continuum excitation,” J. Biomed. Opt. 15(2), 026026 (2010).
[Crossref] [PubMed]

Nature (1)

W. Min, S. Lu, S. Chong, R. Roy, G. R. Holtom, and X. S. Xie, “Imaging chromophores with undetectable fluorescence by stimulated emission microscopy,” Nature 461(7267), 1105–1109 (2009).
[Crossref] [PubMed]

Opt. Express (4)

Opt. Lett. (5)

Phys. Rev. Lett. (1)

A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett. 82(20), 4142 (1999).
[Crossref]

Proc. Natl. Acad. Sci. U.S.A. (1)

C. L. Evans, E. O. Potma, M. Puoris’haag, D. Côté, C. P. Lin, and X. S. Xie, “Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy,” Proc. Natl. Acad. Sci. U.S.A. 102(46), 16807–16812 (2005).
[Crossref] [PubMed]

Science (1)

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, “Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy,” Science 322(5909), 1857–1861 (2008).
[Crossref] [PubMed]

Supplementary Material (1)

» Media 1: MOV (1439 KB)     

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Figures (3)

Fig. 1
Fig. 1 Experimental setup of the 1064 nm time-lens source synchronized to a mode-locked laser. The time-lens source (enclosed by the dotted box) consists of a 1064 nm CW laser diode, 2 phase modulators (2PMs), 2 Mach-Zehnder intensity modulators (MZ1 and MZ2), a fiber circulator, a chirped fiber Bragg grating (CFBG), and Yb3+-doped fiber amplifiers (YDFAs, 2 preamps and 1 power amp). All fiber devices are polarization maintained except for the YDFAs where polarization controllers (PCs) are used to align the polarization states. The RF signals used to drive the 2 PMs and MZ1 are derived from the fast photodetector. MZ2 is driven with an external function generator at 10 MHz for SRS imaging. BS: beam splitter, DC: dichroic mirror, NB: narrow band, BB: broad band. Electrical paths are labeled with dashed lines, and optical paths are labeled with solid lines.

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Fig. 2
Fig. 2 Characterization of the time-lens source. (a) optical spectrum of the time-lens output (blue) and the initial input CW laser (black). (b) cross-correlation trace between the time-lens source and the 130-fs, 77-MHz pulse train (blue) from a mode-locked fiber laser. Inset shows the measured sum-frequency signal at the half maximum of the cross-correlation trace over a time span of 350 seconds (red, same vertical scale). (c) cross-correlation trace between the time-lens source and the 2-ps, 76-MHz Ti:Sa mode-locked laser (blue). Inset shows the measured sum-frequency signal at the half maximum of the cross-correlation trace over a time span of 350 seconds (red). (d) oscilloscope trace of the time-lens source modulated by a 10-MHz square wave (yellow) and the pulses of the Ti:Sa mode-locked laser (purple).

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Fig. 3
Fig. 3 Label-free CRS imaging of tissue samples. (a) SRS image of mouse brain at the CH2 stretching frequency (2845 cm−1), showing individual myelinated neurons. (b) SRS image of mouse skin showing the sebaceous glands with sub-cellular resolution in the viable epidermis. (c) a single frame from a CARS movie acquired at video-rate (30 fps). (d) SRS image of drug penetration of the skin-active ingredient trans-retinol in the stratum corneum obtained at the polyene stretching frequency (1600 cm−1). All scale bars are 25 μm. (Media 1)

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