Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Stimulated Raman scattering microscopy with a robust fibre laser source


Stimulated Raman scattering microscopy allows label-free chemical imaging and has enabled exciting applications in biology, material science and medicine. It provides a major advantage in imaging speed over spontaneous Raman scattering and has improved image contrast and spectral fidelity compared to coherent anti-Stokes Raman scattering. Wider adoption of the technique has, however, been hindered by the need for a costly and environmentally sensitive tunable ultrafast dual-wavelength source. We present the development of an optimized all-fibre laser system based on the optical synchronization of two picosecond power amplifiers. To circumvent the high-frequency laser noise intrinsic to amplified fibre lasers, we have further developed a high-speed noise cancellation system based on voltage-subtraction autobalanced detection. We demonstrate uncompromised imaging performance of our fibre-laser-based stimulated Raman scattering microscope with shot-noise-limited sensitivity and an imaging speed up to 1 frame s−1.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Schematic of the fibre-laser system and SRS microscope.
Figure 2: Characterization of the fibre-laser source.
Figure 3: Autobalanced detection.
Figure 4: SRS spectral imaging with the fibre-laser source.


  1. 1

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

    ADS  Article  Google Scholar 

  2. 2

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

    Article  Google Scholar 

  3. 3

    Ploetz, E., Laimgruber, S., Berner, S., Zinth, W. & Gilch, P. Femtosecond stimulated Raman microscopy. Appl. Phys. B 87, 389–393 (2007).

    ADS  Article  Google Scholar 

  4. 4

    Freudiger, C. W. et al. Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy. Science 322, 1857–1861 (2008).

    ADS  Article  Google Scholar 

  5. 5

    Ozeki, Y., Dake, F., Kajiyama, S., Fukui, K. & Itoh, K. Analysis and experimental assessment of the sensitivity of stimulated Raman scattering microscopy. Opt. Express 17, 3651–3658 (2009).

    ADS  Article  Google Scholar 

  6. 6

    Nandakumar, P., Kovalev, A. & Volkmer, A. Vibrational imaging based on stimulated Raman scattering microscopy. New J. Phys. 11, 033026 (2009).

    ADS  Article  Google Scholar 

  7. 7

    Bloembergen, N. The stimulated Raman effect. Am. J. Phys. 35, 989–1023 (1967).

    ADS  Article  Google Scholar 

  8. 8

    Evans, C. L. et al. Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy. Proc. Natl Acad. Sci. USA 102, 16807–16812 (2005).

    ADS  Article  Google Scholar 

  9. 9

    Saar, B. G. et al. Video-rate molecular imaging in vivo with stimulated Raman scattering. Science 330, 1368–1370 (2010).

    ADS  Article  Google Scholar 

  10. 10

    Ganikhanov, F., Evans, C. L., Saar, B. G. & Xie, X. S. High-sensitivity vibrational imaging with frequency modulation coherent anti-Stokes Raman scattering (FM CARS) microscopy. Opt. Lett. 31, 1872–1874 (2006).

    ADS  Article  Google Scholar 

  11. 11

    Min, W., Freudiger, C. W., Lu, S. & Xie, X. S. Coherent nonlinear optical imaging: beyond fluorescence microscopy. Annu. Rev. Phys. Chem. 62, 507–530 (2011).

    ADS  Article  Google Scholar 

  12. 12

    Owyoung, A. Sensitivity limitations for CW stimulated Raman spectroscopy. Opt. Commun. 22, 323–328 (1977).

    ADS  Article  Google Scholar 

  13. 13

    Ozeki, Y. et al. Stimulated Raman scattering microscope with shot noise limited sensitivity using subharmonically synchronized laser pulses. Opt. Express 18, 13708–13719 (2010).

    ADS  Article  Google Scholar 

  14. 14

    Ji, M. et al. Rapid, label-free detection of brain tumors with stimulated Raman scattering microscopy. Sci. Transl. Med. 5, 201ra119 (2013).

    Article  Google Scholar 

  15. 15

    Lin, C.-Y. et al. Picosecond spectral coherent anti-Stokes Raman scattering imaging with principal component analysis of meibomian glands. J. Biomed. Opt. 16, 021104 (2011).

    ADS  Article  Google Scholar 

  16. 16

    Ozeki, Y. et al. High-speed molecular spectral imaging of tissue with stimulated Raman scattering. Nature Photon. 6, 845–851 (2012).

    ADS  Article  Google Scholar 

  17. 17

    Kong, L. et al. Multicolor stimulated Raman scattering microscopy with a rapidly tunable optical parametric oscillator. Opt. Lett. 38, 145–147 (2013).

    ADS  Article  Google Scholar 

  18. 18

    Ganikhanov, F. et al. Broadly tunable dual-wavelength light source for coherent anti-Stokes Raman scattering microscopy. Opt. Lett. 31, 1292–1294 (2006).

    ADS  Article  Google Scholar 

  19. 19

    Jones, D. J. et al. Synchronization of two passively mode-locked, picosecond lasers within 20 fs for coherent anti-Stokes Raman scattering microscopy. Rev. Sci. Instrum. 73, 2843–2848 (2002).

    ADS  Article  Google Scholar 

  20. 20

    Pegoraro, A. F. et al. Optimally chirped multimodal CARS microscopy based on a single Ti:sapphire oscillator. Opt. Express 17, 2984–2996 (2009).

    ADS  Article  Google Scholar 

  21. 21

    Krauss, G. et al. Compact coherent anti-Stokes Raman scattering microscope based on a picosecond two-color Er:fiber laser system. Opt. Lett. 34, 2847–2849 (2009).

    ADS  Article  Google Scholar 

  22. 22

    Gambetta, A. et al. Fiber-format stimulated-Raman-scattering microscopy from a single laser oscillator. Opt. Lett. 35, 226–228 (2010).

    ADS  Article  Google Scholar 

  23. 23

    Baumgartl, M. et al. All-fiber laser source for CARS microscopy based on fiber optical parametric frequency conversion. Opt. Express 20, 4484–4493 (2012).

    ADS  Article  Google Scholar 

  24. 24

    Lefrancois, S. et al. Fiber four-wave mixing source for coherent anti-Stokes Raman scattering microscopy. Opt. Lett. 37, 1652–1654 (2012).

    ADS  Article  Google Scholar 

  25. 25

    Bégin, S. et al. Coherent anti-Stokes Raman scattering hyperspectral tissue imaging with a wavelength-swept system. Biomed. Opt. Express 2, 1296–1306 (2011).

    Article  Google Scholar 

  26. 26

    Nose, K. et al. Sensitivity enhancement of fiber-laser-based stimulated Raman scattering microscopy by collinear balanced detection technique. Opt. Express 20, 13958–13965 (2012).

    ADS  Article  Google Scholar 

  27. 27

    Hobbs, P. C. D. Shot noise limited optical measurements at baseband with noisy lasers. Proc. SPIE 1376, 216–221 (1991).

    ADS  Article  Google Scholar 

  28. 28

    Hobbs, P. C. D. Building Electro-optical Systems: Making It All Work (Wiley, 2000).

    Book  Google Scholar 

  29. 29

    Kieu, K. & Mansuripur, M. Femtosecond laser pulse generation with a fiber taper embedded in carbon nanotube/polymer composite. Opt. Lett. 32, 2242–2244 (2007).

    ADS  Article  Google Scholar 

  30. 30

    Kieu, K., Jones, J. & Peyghambarian, N. High power femtosecond source near 1 micron based on an all-fiber Er-doped mode-locked laser. Opt. Express 18, 21350–21355 (2010).

    ADS  Article  Google Scholar 

  31. 31

    Andrianov, A., Anashkina, E., Muravyev, S. & Kim, A. All-fiber design of hybrid Er-doped laser/Yb-doped amplifier system for high-power ultrashort pulse generation. Opt. Lett. 35, 3805–3807 (2010).

    ADS  Article  Google Scholar 

  32. 32

    Hopt, A. & Neher, E. Highly nonlinear photodamage in two-photon fluorescence microscopy. Biophys. J. 80, 2029–2036 (2001).

    ADS  Article  Google Scholar 

  33. 33

    Fu, Y., Wang, H., Shi, R. & Cheng, J.-X. Characterization of photodamage in coherent anti-Stokes Raman scattering microscopy. Opt. Express 14, 3942–3951 (2006).

    ADS  Article  Google Scholar 

  34. 34

    Nan, X., Potma, E. O. & Xie, X. S. Nonperturbative chemical imaging of organelle transport in living cells with coherent anti-Stokes Raman scattering microscopy. Biophys. J. 91, 728–735 (2006).

    ADS  Article  Google Scholar 

  35. 35

    Obarski, G. E. & Hale, P. D. How to measure relative intensity noise in lasers. Laser Focus World 35, 273–278 (1999).

    Google Scholar 

Download references


The authors thank P. Hobbs, J. McArthur and J. Trautman for discussions. The authors thank D. Fu and F.-K. Lu for help with sample preparation. This material is based on work supported by the National Science Foundation (NSF; grant no. 1214848 to C.W.F.) and by the National Institutes of Health (NIH; grant no. 5R01EB010244 to X.S.X). This work was performed in part at the Center for Nanoscale Systems (CNS), a member of the National Nanotechnology Infrastructure Network (NNIN), which is supported by the NSF (under grant no. ECS-0335765). CNS is part of Harvard University. KK and NP would like to acknowledge the support from the CIAN NSF ERC under grant #EEC-0812072 and the State of Arizona’s TRIF funding.

Author information




C.W.F. and K.Q.K. designed and characterized the fibre-laser system. W.Y. and G.R.H. designed and characterized the autobalanced detector. C.W.F. and W.Y. performed the imaging experiments. C.W.F., X.S.X., N.P. and K.Q.K. conceived the project and supervised its implementation. C.W.F., W.Y., K.Q.K., N.P. and X.S.X. wrote the manuscript and all authors commented on it.

Corresponding authors

Correspondence to X. Sunney Xie or Khanh Q. Kieu.

Ethics declarations

Competing interests

Harvard University and University of Arizona has filed a patent application based on the current work. C.W.F. and X.S.X. have financial interests in Invenio Imaging Inc. K.Q.K. has financial interests in KPhotonics LLC. Any opinions, findings, conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the NSF or NIH.

Supplementary information

Supplementary information

Supplementary information (PDF 820 kb)

Supplementary information

Supplementary movie (AVI 3134 kb)

Supplementary information

Supplementary movie (AVI 4720 kb)

Supplementary information

Supplementary movie (MOV 901 kb)

Supplementary information

Supplementary movie (MOV 6383 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Freudiger, C., Yang, W., Holtom, G. et al. Stimulated Raman scattering microscopy with a robust fibre laser source. Nature Photon 8, 153–159 (2014).

Download citation

Further reading


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing