Abstract

Stimulated Raman scattering (SRS) microscopy allows label-free chemical imaging based on vibrational spectroscopy. Narrowband excitation with picosecond lasers creates the highest signal levels and enables imaging speeds up to video-rate, but it sacrifices chemical specificity in samples with overlapping bands compared to broadband (multiplex) excitation. We develop a rapidly tunable picosecond optical parametric oscillator with an electro-optical tunable Lyot filter, and demonstrate multicolor SRS microscopy with synchronized line-by-line wavelength tuning to avoid spectral artifacts due to sample movement. We show sensitive imaging of three different kinds of polymer beads and live HeLa cells with moving intracellular lipid droplets.

© 2013 Optical Society of America

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    [CrossRef]
  2. J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, J. Phys. Chem. B 105, 1277 (2001).
    [CrossRef]
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    [CrossRef]
  4. 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, Science 322, 1857 (2008).
    [CrossRef]
  5. Y. Ozeki, F. Dake, S. I. Kajiyama, K. Fukui, and K. Itoh, Opt. Express 17, 3651 (2009).
    [CrossRef]
  6. P. Nandakumar, A. Kovalev, and A. Volkmer, New J. Phys. 11, 033026 (2009).
    [CrossRef]
  7. B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, Science 330, 1368 (2010).
    [CrossRef]
  8. C.-Y. Lin, J. L. Suhalim, C. L. Nien, M. D. Miljkovic, M. Diem, J. V. Jester, and E. O. Potma, J. Biomed. Opt. 16, 021104 (2011).
    [CrossRef]
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2012 (4)

Y. Ozeki, W. Umemura, K. Sumimura, N. Nishizawa, K. Fukui, and K. Itoh, Opt. Lett. 37, 431 (2012).
[CrossRef]

Y. Ozeki, W. Umemura, Y. Otsuka, S. Satoh, H. Hashimoto, K. Sumimura, N. Nishizawa, K. Fukui, and K. Itoh, Nat. Photonics 6, 845 (2012).
[CrossRef]

D. Fu, F.-K. Lu, X. Zhang, C. Freudiger, D. R. Pernik, G. Holtom, and X. S. Xie, J. Am. Chem. Soc. 134, 3623 (2012).
[CrossRef]

F.-K. Lu, M. Ji, D. Fu, X. Ni, C. W. Freudiger, G. Holtom, and X. S. Xie, Mol. Phys. 110, 1927 (2012).
[CrossRef]

2011 (2)

C. W. Freudiger, W. Min, G. R. Holtom, B. Xu, M. Dantus, and X. S. Xie, Nat. Photonics 5, 103 (2011).
[CrossRef]

C.-Y. Lin, J. L. Suhalim, C. L. Nien, M. D. Miljkovic, M. Diem, J. V. Jester, and E. O. Potma, J. Biomed. Opt. 16, 021104 (2011).
[CrossRef]

2010 (1)

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, Science 330, 1368 (2010).
[CrossRef]

2009 (3)

2008 (2)

C. L. Evans and X. S. Xie, Annu. Rev. Phys. Chem. 1, 883 (2008).
[CrossRef]

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, Science 322, 1857 (2008).
[CrossRef]

2007 (1)

E. Ploetz, S. Laimgruber, S. Berner, W. Zinth, and P. Gilch, Appl. Phys. B 87, 389 (2007).
[CrossRef]

2004 (1)

A. Godard and E. Rosencher, IEEE J. Quantum Electron. 40, 784 (2004).
[CrossRef]

2002 (2)

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, J. Phys. Chem. B 106, 8493 (2002).
[CrossRef]

G. W. H. Wurpel, J. M. Schins, and M. Muller, Opt. Lett. 27, 1093 (2002).
[CrossRef]

2001 (1)

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, J. Phys. Chem. B 105, 1277 (2001).
[CrossRef]

1999 (1)

A. Zumbusch, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
[CrossRef]

1997 (1)

Ackermann, C.

Bennett, G. L.

Berner, S.

E. Ploetz, S. Laimgruber, S. Berner, W. Zinth, and P. Gilch, Appl. Phys. B 87, 389 (2007).
[CrossRef]

Book, L. D.

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, J. Phys. Chem. B 106, 8493 (2002).
[CrossRef]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, J. Phys. Chem. B 105, 1277 (2001).
[CrossRef]

Cheng, J. X.

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, J. Phys. Chem. B 106, 8493 (2002).
[CrossRef]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, J. Phys. Chem. B 105, 1277 (2001).
[CrossRef]

Dake, F.

Dantus, M.

C. W. Freudiger, W. Min, G. R. Holtom, B. Xu, M. Dantus, and X. S. Xie, Nat. Photonics 5, 103 (2011).
[CrossRef]

Diem, M.

C.-Y. Lin, J. L. Suhalim, C. L. Nien, M. D. Miljkovic, M. Diem, J. V. Jester, and E. O. Potma, J. Biomed. Opt. 16, 021104 (2011).
[CrossRef]

Evans, C. L.

C. L. Evans and X. S. Xie, Annu. Rev. Phys. Chem. 1, 883 (2008).
[CrossRef]

Ewbank, M. D.

Freudiger, C.

D. Fu, F.-K. Lu, X. Zhang, C. Freudiger, D. R. Pernik, G. Holtom, and X. S. Xie, J. Am. Chem. Soc. 134, 3623 (2012).
[CrossRef]

Freudiger, C. W.

F.-K. Lu, M. Ji, D. Fu, X. Ni, C. W. Freudiger, G. Holtom, and X. S. Xie, Mol. Phys. 110, 1927 (2012).
[CrossRef]

C. W. Freudiger, W. Min, G. R. Holtom, B. Xu, M. Dantus, and X. S. Xie, Nat. Photonics 5, 103 (2011).
[CrossRef]

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, Science 330, 1368 (2010).
[CrossRef]

B. G. Saar, G. R. Holtom, C. W. Freudiger, C. Ackermann, W. Hill, and X. S. Xie, Opt. Express 17, 12532 (2009).
[CrossRef]

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, Science 322, 1857 (2008).
[CrossRef]

Fu, D.

D. Fu, F.-K. Lu, X. Zhang, C. Freudiger, D. R. Pernik, G. Holtom, and X. S. Xie, J. Am. Chem. Soc. 134, 3623 (2012).
[CrossRef]

F.-K. Lu, M. Ji, D. Fu, X. Ni, C. W. Freudiger, G. Holtom, and X. S. Xie, Mol. Phys. 110, 1927 (2012).
[CrossRef]

Fukui, K.

Gilch, P.

E. Ploetz, S. Laimgruber, S. Berner, W. Zinth, and P. Gilch, Appl. Phys. B 87, 389 (2007).
[CrossRef]

Godard, A.

A. Godard and E. Rosencher, IEEE J. Quantum Electron. 40, 784 (2004).
[CrossRef]

Hashimoto, H.

Y. Ozeki, W. Umemura, Y. Otsuka, S. Satoh, H. Hashimoto, K. Sumimura, N. Nishizawa, K. Fukui, and K. Itoh, Nat. Photonics 6, 845 (2012).
[CrossRef]

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, Science 322, 1857 (2008).
[CrossRef]

Hill, W.

Holtom, G.

F.-K. Lu, M. Ji, D. Fu, X. Ni, C. W. Freudiger, G. Holtom, and X. S. Xie, Mol. Phys. 110, 1927 (2012).
[CrossRef]

D. Fu, F.-K. Lu, X. Zhang, C. Freudiger, D. R. Pernik, G. Holtom, and X. S. Xie, J. Am. Chem. Soc. 134, 3623 (2012).
[CrossRef]

Holtom, G. R.

C. W. Freudiger, W. Min, G. R. Holtom, B. Xu, M. Dantus, and X. S. Xie, Nat. Photonics 5, 103 (2011).
[CrossRef]

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, Science 330, 1368 (2010).
[CrossRef]

B. G. Saar, G. R. Holtom, C. W. Freudiger, C. Ackermann, W. Hill, and X. S. Xie, Opt. Express 17, 12532 (2009).
[CrossRef]

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, Science 322, 1857 (2008).
[CrossRef]

A. Zumbusch, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
[CrossRef]

Itoh, K.

Jester, J. V.

C.-Y. Lin, J. L. Suhalim, C. L. Nien, M. D. Miljkovic, M. Diem, J. V. Jester, and E. O. Potma, J. Biomed. Opt. 16, 021104 (2011).
[CrossRef]

Ji, M.

F.-K. Lu, M. Ji, D. Fu, X. Ni, C. W. Freudiger, G. Holtom, and X. S. Xie, Mol. Phys. 110, 1927 (2012).
[CrossRef]

Kajiyama, S. I.

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, Science 322, 1857 (2008).
[CrossRef]

Kovalev, A.

P. Nandakumar, A. Kovalev, and A. Volkmer, New J. Phys. 11, 033026 (2009).
[CrossRef]

Laimgruber, S.

E. Ploetz, S. Laimgruber, S. Berner, W. Zinth, and P. Gilch, Appl. Phys. B 87, 389 (2007).
[CrossRef]

Lin, C.-Y.

C.-Y. Lin, J. L. Suhalim, C. L. Nien, M. D. Miljkovic, M. Diem, J. V. Jester, and E. O. Potma, J. Biomed. Opt. 16, 021104 (2011).
[CrossRef]

Lu, F.-K.

F.-K. Lu, M. Ji, D. Fu, X. Ni, C. W. Freudiger, G. Holtom, and X. S. Xie, Mol. Phys. 110, 1927 (2012).
[CrossRef]

D. Fu, F.-K. Lu, X. Zhang, C. Freudiger, D. R. Pernik, G. Holtom, and X. S. Xie, J. Am. Chem. Soc. 134, 3623 (2012).
[CrossRef]

Lu, S.

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, Science 322, 1857 (2008).
[CrossRef]

Miljkovic, M. D.

C.-Y. Lin, J. L. Suhalim, C. L. Nien, M. D. Miljkovic, M. Diem, J. V. Jester, and E. O. Potma, J. Biomed. Opt. 16, 021104 (2011).
[CrossRef]

Min, W.

C. W. Freudiger, W. Min, G. R. Holtom, B. Xu, M. Dantus, and X. S. Xie, Nat. Photonics 5, 103 (2011).
[CrossRef]

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, Science 322, 1857 (2008).
[CrossRef]

Muller, M.

Nandakumar, P.

P. Nandakumar, A. Kovalev, and A. Volkmer, New J. Phys. 11, 033026 (2009).
[CrossRef]

Ni, X.

F.-K. Lu, M. Ji, D. Fu, X. Ni, C. W. Freudiger, G. Holtom, and X. S. Xie, Mol. Phys. 110, 1927 (2012).
[CrossRef]

Nien, C. L.

C.-Y. Lin, J. L. Suhalim, C. L. Nien, M. D. Miljkovic, M. Diem, J. V. Jester, and E. O. Potma, J. Biomed. Opt. 16, 021104 (2011).
[CrossRef]

Nishizawa, N.

Y. Ozeki, W. Umemura, K. Sumimura, N. Nishizawa, K. Fukui, and K. Itoh, Opt. Lett. 37, 431 (2012).
[CrossRef]

Y. Ozeki, W. Umemura, Y. Otsuka, S. Satoh, H. Hashimoto, K. Sumimura, N. Nishizawa, K. Fukui, and K. Itoh, Nat. Photonics 6, 845 (2012).
[CrossRef]

Otsuka, Y.

Y. Ozeki, W. Umemura, Y. Otsuka, S. Satoh, H. Hashimoto, K. Sumimura, N. Nishizawa, K. Fukui, and K. Itoh, Nat. Photonics 6, 845 (2012).
[CrossRef]

Ozeki, Y.

Pernik, D. R.

D. Fu, F.-K. Lu, X. Zhang, C. Freudiger, D. R. Pernik, G. Holtom, and X. S. Xie, J. Am. Chem. Soc. 134, 3623 (2012).
[CrossRef]

Ploetz, E.

E. Ploetz, S. Laimgruber, S. Berner, W. Zinth, and P. Gilch, Appl. Phys. B 87, 389 (2007).
[CrossRef]

Potma, E. O.

C.-Y. Lin, J. L. Suhalim, C. L. Nien, M. D. Miljkovic, M. Diem, J. V. Jester, and E. O. Potma, J. Biomed. Opt. 16, 021104 (2011).
[CrossRef]

Reichman, J.

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, Science 330, 1368 (2010).
[CrossRef]

Rosencher, E.

A. Godard and E. Rosencher, IEEE J. Quantum Electron. 40, 784 (2004).
[CrossRef]

Rosker, M. J.

Saar, B. G.

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, Science 330, 1368 (2010).
[CrossRef]

B. G. Saar, G. R. Holtom, C. W. Freudiger, C. Ackermann, W. Hill, and X. S. Xie, Opt. Express 17, 12532 (2009).
[CrossRef]

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, Science 322, 1857 (2008).
[CrossRef]

Satoh, S.

Y. Ozeki, W. Umemura, Y. Otsuka, S. Satoh, H. Hashimoto, K. Sumimura, N. Nishizawa, K. Fukui, and K. Itoh, Nat. Photonics 6, 845 (2012).
[CrossRef]

Schins, J. M.

Stanley, C. M.

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, Science 330, 1368 (2010).
[CrossRef]

Suhalim, J. L.

C.-Y. Lin, J. L. Suhalim, C. L. Nien, M. D. Miljkovic, M. Diem, J. V. Jester, and E. O. Potma, J. Biomed. Opt. 16, 021104 (2011).
[CrossRef]

Sumimura, K.

Y. Ozeki, W. Umemura, K. Sumimura, N. Nishizawa, K. Fukui, and K. Itoh, Opt. Lett. 37, 431 (2012).
[CrossRef]

Y. Ozeki, W. Umemura, Y. Otsuka, S. Satoh, H. Hashimoto, K. Sumimura, N. Nishizawa, K. Fukui, and K. Itoh, Nat. Photonics 6, 845 (2012).
[CrossRef]

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, Science 322, 1857 (2008).
[CrossRef]

Umemura, W.

Y. Ozeki, W. Umemura, K. Sumimura, N. Nishizawa, K. Fukui, and K. Itoh, Opt. Lett. 37, 431 (2012).
[CrossRef]

Y. Ozeki, W. Umemura, Y. Otsuka, S. Satoh, H. Hashimoto, K. Sumimura, N. Nishizawa, K. Fukui, and K. Itoh, Nat. Photonics 6, 845 (2012).
[CrossRef]

Volkmer, A.

P. Nandakumar, A. Kovalev, and A. Volkmer, New J. Phys. 11, 033026 (2009).
[CrossRef]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, J. Phys. Chem. B 106, 8493 (2002).
[CrossRef]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, J. Phys. Chem. B 105, 1277 (2001).
[CrossRef]

Wurpel, G. W. H.

Xie, X. S.

F.-K. Lu, M. Ji, D. Fu, X. Ni, C. W. Freudiger, G. Holtom, and X. S. Xie, Mol. Phys. 110, 1927 (2012).
[CrossRef]

D. Fu, F.-K. Lu, X. Zhang, C. Freudiger, D. R. Pernik, G. Holtom, and X. S. Xie, J. Am. Chem. Soc. 134, 3623 (2012).
[CrossRef]

C. W. Freudiger, W. Min, G. R. Holtom, B. Xu, M. Dantus, and X. S. Xie, Nat. Photonics 5, 103 (2011).
[CrossRef]

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, Science 330, 1368 (2010).
[CrossRef]

B. G. Saar, G. R. Holtom, C. W. Freudiger, C. Ackermann, W. Hill, and X. S. Xie, Opt. Express 17, 12532 (2009).
[CrossRef]

C. L. Evans and X. S. Xie, Annu. Rev. Phys. Chem. 1, 883 (2008).
[CrossRef]

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, Science 322, 1857 (2008).
[CrossRef]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, J. Phys. Chem. B 106, 8493 (2002).
[CrossRef]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, J. Phys. Chem. B 105, 1277 (2001).
[CrossRef]

A. Zumbusch, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
[CrossRef]

Xu, B.

C. W. Freudiger, W. Min, G. R. Holtom, B. Xu, M. Dantus, and X. S. Xie, Nat. Photonics 5, 103 (2011).
[CrossRef]

Zhang, X.

D. Fu, F.-K. Lu, X. Zhang, C. Freudiger, D. R. Pernik, G. Holtom, and X. S. Xie, J. Am. Chem. Soc. 134, 3623 (2012).
[CrossRef]

Zinth, W.

E. Ploetz, S. Laimgruber, S. Berner, W. Zinth, and P. Gilch, Appl. Phys. B 87, 389 (2007).
[CrossRef]

Zumbusch, A.

A. Zumbusch, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
[CrossRef]

Annu. Rev. Phys. Chem. (1)

C. L. Evans and X. S. Xie, Annu. Rev. Phys. Chem. 1, 883 (2008).
[CrossRef]

Appl. Phys. B (1)

E. Ploetz, S. Laimgruber, S. Berner, W. Zinth, and P. Gilch, Appl. Phys. B 87, 389 (2007).
[CrossRef]

IEEE J. Quantum Electron. (1)

A. Godard and E. Rosencher, IEEE J. Quantum Electron. 40, 784 (2004).
[CrossRef]

J. Am. Chem. Soc. (1)

D. Fu, F.-K. Lu, X. Zhang, C. Freudiger, D. R. Pernik, G. Holtom, and X. S. Xie, J. Am. Chem. Soc. 134, 3623 (2012).
[CrossRef]

J. Biomed. Opt. (1)

C.-Y. Lin, J. L. Suhalim, C. L. Nien, M. D. Miljkovic, M. Diem, J. V. Jester, and E. O. Potma, J. Biomed. Opt. 16, 021104 (2011).
[CrossRef]

J. Opt. Soc. Am. B (1)

J. Phys. Chem. B (2)

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, J. Phys. Chem. B 105, 1277 (2001).
[CrossRef]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, J. Phys. Chem. B 106, 8493 (2002).
[CrossRef]

Mol. Phys. (1)

F.-K. Lu, M. Ji, D. Fu, X. Ni, C. W. Freudiger, G. Holtom, and X. S. Xie, Mol. Phys. 110, 1927 (2012).
[CrossRef]

Nat. Photonics (2)

Y. Ozeki, W. Umemura, Y. Otsuka, S. Satoh, H. Hashimoto, K. Sumimura, N. Nishizawa, K. Fukui, and K. Itoh, Nat. Photonics 6, 845 (2012).
[CrossRef]

C. W. Freudiger, W. Min, G. R. Holtom, B. Xu, M. Dantus, and X. S. Xie, Nat. Photonics 5, 103 (2011).
[CrossRef]

New J. Phys. (1)

P. Nandakumar, A. Kovalev, and A. Volkmer, New J. Phys. 11, 033026 (2009).
[CrossRef]

Opt. Express (2)

Opt. Lett. (2)

Phys. Rev. Lett. (1)

A. Zumbusch, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
[CrossRef]

Science (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, Science 322, 1857 (2008).
[CrossRef]

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, Science 330, 1368 (2010).
[CrossRef]

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Figures (3)

Fig. 1.
Fig. 1.

(a) Schematic of the fast tuning OPO. DM: dichroic mirror, M: mirror, G: Glan-Brewster prism, W: waveplate, EO: electro-optical crystal, LBO: lithium triborate crystal. (b) Tuning characteristic of the OPO with an electric-optical tunable Lyot filter. Dots show average values, and the error bars show the standard deviations after 100 repetitions.

Fig. 2.
Fig. 2.

(a) Spontaneous Raman spectra of melamine, PMMA, and polystyrene beads. (b–d) Raw images taken at 3054, 2979, 2959cm1. (e) Reconstructed SRS image (Red: 2 μm polystyrene, Green: 1–10 μm PMMA, Blue: 2.9 μm.

Fig. 3.
Fig. 3.

(a) Spontaneous Raman spectra of protein and lipid. (b) Reconstructed SRS image by line-by-line wavelength tuning, free of spectral artifacts due to lipid droplet driftings. (c) Reconstructed SRS image by frame-by-frame wavelength tuning. Two arrows point to artifacts due to lipid droplet driftings. Red: protein, green: lipid. Both images are 460×318 pixels. The scale bar is 10 μm (60× water objective lens used).

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