Abstract

Spontaneous Raman microscopy is a potentially useful technique for imaging living cells, tissue and small animals without any probe or dye labeling. We have developed a spontaneous Raman imaging system in wide-field view, which we term ‘light sheet-excited direct Raman spectroscopy’ (LSDRS). This system, which we reported previously, consists of a background-free electrically tunable Ti:Sapphire laser (BF-ETL), a cylindrical lens, a CCD camera, and a narrow bandpass filter. Here, we have adapted the LSDRS system for microscopy systems, such as single-plane illumination microscopy (SPIM) for biomedical applications, and demonstrated spontaneous Raman imaging of a living fish. The results suggest that our Raman microscopy system enables investigation of the differentiation process and mechanism of iridocytes during development. This is the first report in which Raman imaging of a living animal was successfully demonstrated by spontaneous Raman scattering signals, but not nonlinear Raman effects such as CARS and SRS.

© 2012 OSA

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    [CrossRef]
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    [CrossRef]
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]

2012

J. Huisken, “Slicing embryos gently with laser light sheets,” Bioessays34(5), 406–411 (2012).
[CrossRef] [PubMed]

2011

M. D. Morris and G. S. Mandair, “Raman assessment of bone quality,” Clin. Orthop. Relat. Res.469(8), 2160–2169 (2011).
[CrossRef] [PubMed]

B. G. Saar, L. R. Contreras-Rojas, X. S. Xie, and R. H. Guy, “Imaging drug delivery to skin with stimulated Raman scattering microscopy,” Mol. Pharm.8(3), 969–975 (2011).
[CrossRef] [PubMed]

J. P. Pezacki, J. A. Blake, D. C. Danielson, D. C. Kennedy, R. K. Lyn, and R. Singaravelu, “Chemical contrast for imaging living systems: molecular vibrations drive CARS microscopy,” Nat. Chem. Biol.7(3), 137–145 (2011).
[CrossRef] [PubMed]

L. Gao, H. Zhou, M. J. Thrall, F. Li, Y. Yang, Z. Wang, P. Luo, K. K. Wong, G. S. Palapattu, and S. T. Wong, “Label-free high-resolution imaging of prostate glands and cavernous nerves using coherent anti-Stokes Raman scattering microscopy,” Biomed. Opt. Express2(4), 915–926 (2011).
[CrossRef] [PubMed]

2010

J. Horsnell, P. Stonelake, J. Christie-Brown, G. Shetty, J. Hutchings, C. Kendall, and N. Stone, “Raman spectroscopy--a new method for the intra-operative assessment of axillary lymph nodes,” Analyst (Lond.)135(12), 3042–3047 (2010).
[CrossRef] [PubMed]

I. Barman, K. M. Tan, and G. P. Singh, “Optical sectioning using single-plane-illumination Raman imaging,” J. Raman Spectrosc.41(10), 1099–1101 (2010).
[CrossRef]

Y. Zhang, H. Hong, and W. Cai, “Imaging with Raman spectroscopy,” Curr. Pharm. Biotechnol.11(6), 654–661 (2010).
[CrossRef] [PubMed]

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, “Video-rate molecular imaging in vivo with stimulated Raman scattering,” Science330(6009), 1368–1370 (2010).
[CrossRef] [PubMed]

2009

C. Krafft, B. Dietzek, and J. Popp, “Raman and CARS microspectroscopy of cells and tissues,” Analyst (Lond.)134(6), 1046–1057 (2009).
[CrossRef] [PubMed]

S. Wachsmann-Hogiu, T. Weeks, and T. Huser, “Chemical analysis in vivo and in vitro by Raman spectroscopy--from single cells to humans,” Curr. Opin. Biotechnol.20(1), 63–73 (2009).
[CrossRef] [PubMed]

Y. Kamei, M. Suzuki, K. Watanabe, K. Fujimori, T. Kawasaki, T. Deguchi, Y. Yoneda, T. Todo, S. Takagi, T. Funatsu, and S. Yuba, “Infrared laser-mediated gene induction in targeted single cells in vivo,” Nat. Methods6(1), 79–81 (2009).
[CrossRef] [PubMed]

T. Minamikawa, M. Hashimoto, K. Fujita, S. Kawata, and T. Araki, “Multi-focus excitation coherent anti-Stokes Raman scattering (CARS) microscopy and its applications for real-time imaging,” Opt. Express17(12), 9526–9536 (2009).
[CrossRef] [PubMed]

Y. Oshima, C. Furihata, and H. Sato, “Light sheet direct Raman imaging technique for observation of mixing of solvents,” Appl. Spectrosc.63(10), 1115–1120 (2009).
[CrossRef] [PubMed]

J. Y. Lee, S. H. Kim, D. W. Moon, and E. S. Lee, “Three-color multiplex CARS for fast imaging and microspectroscopy in the entire CHn stretching vibrational region,” Opt. Express17(25), 22281–22295 (2009).
[CrossRef] [PubMed]

2008

K. Hamada, K. Fujita, N. I. Smith, M. Kobayashi, Y. Inouye, and S. Kawata, “Raman microscopy for dynamic molecular imaging of living cells,” J. Biomed. Opt.13(4), 044027 (2008).
[CrossRef] [PubMed]

K. Fujita and N. I. Smith, “Label-free molecular imaging of living cells,” Mol. Cells26(6), 530–535 (2008).
[PubMed]

P. J. Keller and E. H. Stelzer, “Quantitative in vivo imaging of entire embryos with Digital Scanned Laser Light Sheet Fluorescence Microscopy,” Curr. Opin. Neurobiol.18(6), 624–632 (2008).
[CrossRef] [PubMed]

P. J. Keller, A. D. Schmidt, J. Wittbrodt, and E. H. Stelzer, “Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy,” Science322(5904), 1065–1069 (2008).
[CrossRef] [PubMed]

2007

2006

C. J. Engelbrecht and E. H. Stelzer, “Resolution enhancement in a light-sheet-based microscope (SPIM),” Opt. Lett.31(10), 1477–1479 (2006).
[CrossRef] [PubMed]

S. Thawornchinsombut, J. W. Park, G. Meng, and E. C. Li-Chan, “Raman spectroscopy determines structural changes associated with gelation properties of fish proteins recovered at alkaline pH,” J. Agric. Food Chem.54(6), 2178–2187 (2006).
[CrossRef] [PubMed]

2005

M. Lynn Lamoreux, R. N. Kelsh, Y. Wakamatsu, and K. Ozato, “Pigment pattern formation in the medaka embryo,” Pigment Cell Res.18(2), 64–73 (2005).
[CrossRef] [PubMed]

G. N. Ten, T. G. Bourova, and V. I. Baranov, “Calculation of Raman spectra and structure elucidation of guanine in the polycrystalline state and in aqueous solution,” J. Struct. Chem.46(6), 998–1005 (2005).
[CrossRef]

2004

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305(5686), 1007–1009 (2004).
[CrossRef] [PubMed]

2002

2000

Araki, T.

Baranov, V. I.

G. N. Ten, T. G. Bourova, and V. I. Baranov, “Calculation of Raman spectra and structure elucidation of guanine in the polycrystalline state and in aqueous solution,” J. Struct. Chem.46(6), 998–1005 (2005).
[CrossRef]

Barman, I.

I. Barman, K. M. Tan, and G. P. Singh, “Optical sectioning using single-plane-illumination Raman imaging,” J. Raman Spectrosc.41(10), 1099–1101 (2010).
[CrossRef]

Blake, J. A.

J. P. Pezacki, J. A. Blake, D. C. Danielson, D. C. Kennedy, R. K. Lyn, and R. Singaravelu, “Chemical contrast for imaging living systems: molecular vibrations drive CARS microscopy,” Nat. Chem. Biol.7(3), 137–145 (2011).
[CrossRef] [PubMed]

Bourova, T. G.

G. N. Ten, T. G. Bourova, and V. I. Baranov, “Calculation of Raman spectra and structure elucidation of guanine in the polycrystalline state and in aqueous solution,” J. Struct. Chem.46(6), 998–1005 (2005).
[CrossRef]

Cai, W.

Y. Zhang, H. Hong, and W. Cai, “Imaging with Raman spectroscopy,” Curr. Pharm. Biotechnol.11(6), 654–661 (2010).
[CrossRef] [PubMed]

Christie-Brown, J.

J. Horsnell, P. Stonelake, J. Christie-Brown, G. Shetty, J. Hutchings, C. Kendall, and N. Stone, “Raman spectroscopy--a new method for the intra-operative assessment of axillary lymph nodes,” Analyst (Lond.)135(12), 3042–3047 (2010).
[CrossRef] [PubMed]

Colombelli, J.

Contreras-Rojas, L. R.

B. G. Saar, L. R. Contreras-Rojas, X. S. Xie, and R. H. Guy, “Imaging drug delivery to skin with stimulated Raman scattering microscopy,” Mol. Pharm.8(3), 969–975 (2011).
[CrossRef] [PubMed]

Danielson, D. C.

J. P. Pezacki, J. A. Blake, D. C. Danielson, D. C. Kennedy, R. K. Lyn, and R. Singaravelu, “Chemical contrast for imaging living systems: molecular vibrations drive CARS microscopy,” Nat. Chem. Biol.7(3), 137–145 (2011).
[CrossRef] [PubMed]

Deguchi, T.

Y. Kamei, M. Suzuki, K. Watanabe, K. Fujimori, T. Kawasaki, T. Deguchi, Y. Yoneda, T. Todo, S. Takagi, T. Funatsu, and S. Yuba, “Infrared laser-mediated gene induction in targeted single cells in vivo,” Nat. Methods6(1), 79–81 (2009).
[CrossRef] [PubMed]

Del Bene, F.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305(5686), 1007–1009 (2004).
[CrossRef] [PubMed]

Dietzek, B.

C. Krafft, B. Dietzek, and J. Popp, “Raman and CARS microspectroscopy of cells and tissues,” Analyst (Lond.)134(6), 1046–1057 (2009).
[CrossRef] [PubMed]

Engelbrecht, C. J.

Freudiger, C. W.

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, “Video-rate molecular imaging in vivo with stimulated Raman scattering,” Science330(6009), 1368–1370 (2010).
[CrossRef] [PubMed]

Fujimori, K.

Y. Kamei, M. Suzuki, K. Watanabe, K. Fujimori, T. Kawasaki, T. Deguchi, Y. Yoneda, T. Todo, S. Takagi, T. Funatsu, and S. Yuba, “Infrared laser-mediated gene induction in targeted single cells in vivo,” Nat. Methods6(1), 79–81 (2009).
[CrossRef] [PubMed]

Fujita, K.

T. Minamikawa, M. Hashimoto, K. Fujita, S. Kawata, and T. Araki, “Multi-focus excitation coherent anti-Stokes Raman scattering (CARS) microscopy and its applications for real-time imaging,” Opt. Express17(12), 9526–9536 (2009).
[CrossRef] [PubMed]

K. Hamada, K. Fujita, N. I. Smith, M. Kobayashi, Y. Inouye, and S. Kawata, “Raman microscopy for dynamic molecular imaging of living cells,” J. Biomed. Opt.13(4), 044027 (2008).
[CrossRef] [PubMed]

K. Fujita and N. I. Smith, “Label-free molecular imaging of living cells,” Mol. Cells26(6), 530–535 (2008).
[PubMed]

Funatsu, T.

Y. Kamei, M. Suzuki, K. Watanabe, K. Fujimori, T. Kawasaki, T. Deguchi, Y. Yoneda, T. Todo, S. Takagi, T. Funatsu, and S. Yuba, “Infrared laser-mediated gene induction in targeted single cells in vivo,” Nat. Methods6(1), 79–81 (2009).
[CrossRef] [PubMed]

Furihata, C.

Gao, L.

Greger, K.

Guy, R. H.

B. G. Saar, L. R. Contreras-Rojas, X. S. Xie, and R. H. Guy, “Imaging drug delivery to skin with stimulated Raman scattering microscopy,” Mol. Pharm.8(3), 969–975 (2011).
[CrossRef] [PubMed]

Hamada, K.

K. Hamada, K. Fujita, N. I. Smith, M. Kobayashi, Y. Inouye, and S. Kawata, “Raman microscopy for dynamic molecular imaging of living cells,” J. Biomed. Opt.13(4), 044027 (2008).
[CrossRef] [PubMed]

Hashimoto, M.

Holtom, G. R.

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, “Video-rate molecular imaging in vivo with stimulated Raman scattering,” Science330(6009), 1368–1370 (2010).
[CrossRef] [PubMed]

Hong, H.

Y. Zhang, H. Hong, and W. Cai, “Imaging with Raman spectroscopy,” Curr. Pharm. Biotechnol.11(6), 654–661 (2010).
[CrossRef] [PubMed]

Horsnell, J.

J. Horsnell, P. Stonelake, J. Christie-Brown, G. Shetty, J. Hutchings, C. Kendall, and N. Stone, “Raman spectroscopy--a new method for the intra-operative assessment of axillary lymph nodes,” Analyst (Lond.)135(12), 3042–3047 (2010).
[CrossRef] [PubMed]

Huisken, J.

J. Huisken, “Slicing embryos gently with laser light sheets,” Bioessays34(5), 406–411 (2012).
[CrossRef] [PubMed]

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305(5686), 1007–1009 (2004).
[CrossRef] [PubMed]

Huser, T.

S. Wachsmann-Hogiu, T. Weeks, and T. Huser, “Chemical analysis in vivo and in vitro by Raman spectroscopy--from single cells to humans,” Curr. Opin. Biotechnol.20(1), 63–73 (2009).
[CrossRef] [PubMed]

Hutchings, J.

J. Horsnell, P. Stonelake, J. Christie-Brown, G. Shetty, J. Hutchings, C. Kendall, and N. Stone, “Raman spectroscopy--a new method for the intra-operative assessment of axillary lymph nodes,” Analyst (Lond.)135(12), 3042–3047 (2010).
[CrossRef] [PubMed]

Inouye, Y.

K. Hamada, K. Fujita, N. I. Smith, M. Kobayashi, Y. Inouye, and S. Kawata, “Raman microscopy for dynamic molecular imaging of living cells,” J. Biomed. Opt.13(4), 044027 (2008).
[CrossRef] [PubMed]

Kamei, Y.

Y. Kamei, M. Suzuki, K. Watanabe, K. Fujimori, T. Kawasaki, T. Deguchi, Y. Yoneda, T. Todo, S. Takagi, T. Funatsu, and S. Yuba, “Infrared laser-mediated gene induction in targeted single cells in vivo,” Nat. Methods6(1), 79–81 (2009).
[CrossRef] [PubMed]

Kawasaki, T.

Y. Kamei, M. Suzuki, K. Watanabe, K. Fujimori, T. Kawasaki, T. Deguchi, Y. Yoneda, T. Todo, S. Takagi, T. Funatsu, and S. Yuba, “Infrared laser-mediated gene induction in targeted single cells in vivo,” Nat. Methods6(1), 79–81 (2009).
[CrossRef] [PubMed]

Kawata, S.

T. Minamikawa, M. Hashimoto, K. Fujita, S. Kawata, and T. Araki, “Multi-focus excitation coherent anti-Stokes Raman scattering (CARS) microscopy and its applications for real-time imaging,” Opt. Express17(12), 9526–9536 (2009).
[CrossRef] [PubMed]

K. Hamada, K. Fujita, N. I. Smith, M. Kobayashi, Y. Inouye, and S. Kawata, “Raman microscopy for dynamic molecular imaging of living cells,” J. Biomed. Opt.13(4), 044027 (2008).
[CrossRef] [PubMed]

Keller, P. J.

P. J. Keller and E. H. Stelzer, “Quantitative in vivo imaging of entire embryos with Digital Scanned Laser Light Sheet Fluorescence Microscopy,” Curr. Opin. Neurobiol.18(6), 624–632 (2008).
[CrossRef] [PubMed]

P. J. Keller, A. D. Schmidt, J. Wittbrodt, and E. H. Stelzer, “Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy,” Science322(5904), 1065–1069 (2008).
[CrossRef] [PubMed]

Kelsh, R. N.

M. Lynn Lamoreux, R. N. Kelsh, Y. Wakamatsu, and K. Ozato, “Pigment pattern formation in the medaka embryo,” Pigment Cell Res.18(2), 64–73 (2005).
[CrossRef] [PubMed]

Kendall, C.

J. Horsnell, P. Stonelake, J. Christie-Brown, G. Shetty, J. Hutchings, C. Kendall, and N. Stone, “Raman spectroscopy--a new method for the intra-operative assessment of axillary lymph nodes,” Analyst (Lond.)135(12), 3042–3047 (2010).
[CrossRef] [PubMed]

Kennedy, D. C.

J. P. Pezacki, J. A. Blake, D. C. Danielson, D. C. Kennedy, R. K. Lyn, and R. Singaravelu, “Chemical contrast for imaging living systems: molecular vibrations drive CARS microscopy,” Nat. Chem. Biol.7(3), 137–145 (2011).
[CrossRef] [PubMed]

Kim, S. H.

Kobayashi, M.

K. Hamada, K. Fujita, N. I. Smith, M. Kobayashi, Y. Inouye, and S. Kawata, “Raman microscopy for dynamic molecular imaging of living cells,” J. Biomed. Opt.13(4), 044027 (2008).
[CrossRef] [PubMed]

Krafft, C.

C. Krafft, B. Dietzek, and J. Popp, “Raman and CARS microspectroscopy of cells and tissues,” Analyst (Lond.)134(6), 1046–1057 (2009).
[CrossRef] [PubMed]

Krzic, U.

Lee, E. S.

Lee, J. Y.

Li, F.

Li-Chan, E. C.

S. Thawornchinsombut, J. W. Park, G. Meng, and E. C. Li-Chan, “Raman spectroscopy determines structural changes associated with gelation properties of fish proteins recovered at alkaline pH,” J. Agric. Food Chem.54(6), 2178–2187 (2006).
[CrossRef] [PubMed]

Ling, M. Y.

Luo, P.

Lyn, R. K.

J. P. Pezacki, J. A. Blake, D. C. Danielson, D. C. Kennedy, R. K. Lyn, and R. Singaravelu, “Chemical contrast for imaging living systems: molecular vibrations drive CARS microscopy,” Nat. Chem. Biol.7(3), 137–145 (2011).
[CrossRef] [PubMed]

Lynn Lamoreux, M.

M. Lynn Lamoreux, R. N. Kelsh, Y. Wakamatsu, and K. Ozato, “Pigment pattern formation in the medaka embryo,” Pigment Cell Res.18(2), 64–73 (2005).
[CrossRef] [PubMed]

Mandair, G. S.

M. D. Morris and G. S. Mandair, “Raman assessment of bone quality,” Clin. Orthop. Relat. Res.469(8), 2160–2169 (2011).
[CrossRef] [PubMed]

Meng, G.

S. Thawornchinsombut, J. W. Park, G. Meng, and E. C. Li-Chan, “Raman spectroscopy determines structural changes associated with gelation properties of fish proteins recovered at alkaline pH,” J. Agric. Food Chem.54(6), 2178–2187 (2006).
[CrossRef] [PubMed]

Minamikawa, T.

Moon, D. W.

Morris, M. D.

M. D. Morris and G. S. Mandair, “Raman assessment of bone quality,” Clin. Orthop. Relat. Res.469(8), 2160–2169 (2011).
[CrossRef] [PubMed]

Movasaghi, Z.

Z. Movasaghi, S. Rehman, and I. U. Rehman, “Raman spectroscopy of biological tissues,” Appl. Spectrosc. Rev.42(5), 493–541 (2007).
[CrossRef]

Oshima, Y.

Ozato, K.

M. Lynn Lamoreux, R. N. Kelsh, Y. Wakamatsu, and K. Ozato, “Pigment pattern formation in the medaka embryo,” Pigment Cell Res.18(2), 64–73 (2005).
[CrossRef] [PubMed]

Palapattu, G. S.

Park, J. W.

S. Thawornchinsombut, J. W. Park, G. Meng, and E. C. Li-Chan, “Raman spectroscopy determines structural changes associated with gelation properties of fish proteins recovered at alkaline pH,” J. Agric. Food Chem.54(6), 2178–2187 (2006).
[CrossRef] [PubMed]

Pezacki, J. P.

J. P. Pezacki, J. A. Blake, D. C. Danielson, D. C. Kennedy, R. K. Lyn, and R. Singaravelu, “Chemical contrast for imaging living systems: molecular vibrations drive CARS microscopy,” Nat. Chem. Biol.7(3), 137–145 (2011).
[CrossRef] [PubMed]

Popp, J.

C. Krafft, B. Dietzek, and J. Popp, “Raman and CARS microspectroscopy of cells and tissues,” Analyst (Lond.)134(6), 1046–1057 (2009).
[CrossRef] [PubMed]

Rehman, I. U.

Z. Movasaghi, S. Rehman, and I. U. Rehman, “Raman spectroscopy of biological tissues,” Appl. Spectrosc. Rev.42(5), 493–541 (2007).
[CrossRef]

Rehman, S.

Z. Movasaghi, S. Rehman, and I. U. Rehman, “Raman spectroscopy of biological tissues,” Appl. Spectrosc. Rev.42(5), 493–541 (2007).
[CrossRef]

Reichman, J.

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, “Video-rate molecular imaging in vivo with stimulated Raman scattering,” Science330(6009), 1368–1370 (2010).
[CrossRef] [PubMed]

Reynaud, E. G.

Saar, B. G.

B. G. Saar, L. R. Contreras-Rojas, X. S. Xie, and R. H. Guy, “Imaging drug delivery to skin with stimulated Raman scattering microscopy,” Mol. Pharm.8(3), 969–975 (2011).
[CrossRef] [PubMed]

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, “Video-rate molecular imaging in vivo with stimulated Raman scattering,” Science330(6009), 1368–1370 (2010).
[CrossRef] [PubMed]

Sato, H.

Schmidt, A. D.

P. J. Keller, A. D. Schmidt, J. Wittbrodt, and E. H. Stelzer, “Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy,” Science322(5904), 1065–1069 (2008).
[CrossRef] [PubMed]

Shetty, G.

J. Horsnell, P. Stonelake, J. Christie-Brown, G. Shetty, J. Hutchings, C. Kendall, and N. Stone, “Raman spectroscopy--a new method for the intra-operative assessment of axillary lymph nodes,” Analyst (Lond.)135(12), 3042–3047 (2010).
[CrossRef] [PubMed]

Singaravelu, R.

J. P. Pezacki, J. A. Blake, D. C. Danielson, D. C. Kennedy, R. K. Lyn, and R. Singaravelu, “Chemical contrast for imaging living systems: molecular vibrations drive CARS microscopy,” Nat. Chem. Biol.7(3), 137–145 (2011).
[CrossRef] [PubMed]

Singh, G. P.

I. Barman, K. M. Tan, and G. P. Singh, “Optical sectioning using single-plane-illumination Raman imaging,” J. Raman Spectrosc.41(10), 1099–1101 (2010).
[CrossRef]

Smith, N. I.

K. Fujita and N. I. Smith, “Label-free molecular imaging of living cells,” Mol. Cells26(6), 530–535 (2008).
[PubMed]

K. Hamada, K. Fujita, N. I. Smith, M. Kobayashi, Y. Inouye, and S. Kawata, “Raman microscopy for dynamic molecular imaging of living cells,” J. Biomed. Opt.13(4), 044027 (2008).
[CrossRef] [PubMed]

Stanley, C. M.

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, “Video-rate molecular imaging in vivo with stimulated Raman scattering,” Science330(6009), 1368–1370 (2010).
[CrossRef] [PubMed]

Stelzer, E. H.

P. J. Keller and E. H. Stelzer, “Quantitative in vivo imaging of entire embryos with Digital Scanned Laser Light Sheet Fluorescence Microscopy,” Curr. Opin. Neurobiol.18(6), 624–632 (2008).
[CrossRef] [PubMed]

P. J. Keller, A. D. Schmidt, J. Wittbrodt, and E. H. Stelzer, “Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy,” Science322(5904), 1065–1069 (2008).
[CrossRef] [PubMed]

C. J. Engelbrecht, K. Greger, E. G. Reynaud, U. Krzic, J. Colombelli, and E. H. Stelzer, “Three-dimensional laser microsurgery in light-sheet based microscopy (SPIM),” Opt. Express15(10), 6420–6430 (2007).
[CrossRef] [PubMed]

C. J. Engelbrecht and E. H. Stelzer, “Resolution enhancement in a light-sheet-based microscope (SPIM),” Opt. Lett.31(10), 1477–1479 (2006).
[CrossRef] [PubMed]

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305(5686), 1007–1009 (2004).
[CrossRef] [PubMed]

Stone, N.

J. Horsnell, P. Stonelake, J. Christie-Brown, G. Shetty, J. Hutchings, C. Kendall, and N. Stone, “Raman spectroscopy--a new method for the intra-operative assessment of axillary lymph nodes,” Analyst (Lond.)135(12), 3042–3047 (2010).
[CrossRef] [PubMed]

Stonelake, P.

J. Horsnell, P. Stonelake, J. Christie-Brown, G. Shetty, J. Hutchings, C. Kendall, and N. Stone, “Raman spectroscopy--a new method for the intra-operative assessment of axillary lymph nodes,” Analyst (Lond.)135(12), 3042–3047 (2010).
[CrossRef] [PubMed]

Suzuki, M.

Y. Kamei, M. Suzuki, K. Watanabe, K. Fujimori, T. Kawasaki, T. Deguchi, Y. Yoneda, T. Todo, S. Takagi, T. Funatsu, and S. Yuba, “Infrared laser-mediated gene induction in targeted single cells in vivo,” Nat. Methods6(1), 79–81 (2009).
[CrossRef] [PubMed]

Swoger, J.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305(5686), 1007–1009 (2004).
[CrossRef] [PubMed]

Takagi, S.

Y. Kamei, M. Suzuki, K. Watanabe, K. Fujimori, T. Kawasaki, T. Deguchi, Y. Yoneda, T. Todo, S. Takagi, T. Funatsu, and S. Yuba, “Infrared laser-mediated gene induction in targeted single cells in vivo,” Nat. Methods6(1), 79–81 (2009).
[CrossRef] [PubMed]

Tan, K. M.

I. Barman, K. M. Tan, and G. P. Singh, “Optical sectioning using single-plane-illumination Raman imaging,” J. Raman Spectrosc.41(10), 1099–1101 (2010).
[CrossRef]

Tashiro, H.

Ten, G. N.

G. N. Ten, T. G. Bourova, and V. I. Baranov, “Calculation of Raman spectra and structure elucidation of guanine in the polycrystalline state and in aqueous solution,” J. Struct. Chem.46(6), 998–1005 (2005).
[CrossRef]

Thawornchinsombut, S.

S. Thawornchinsombut, J. W. Park, G. Meng, and E. C. Li-Chan, “Raman spectroscopy determines structural changes associated with gelation properties of fish proteins recovered at alkaline pH,” J. Agric. Food Chem.54(6), 2178–2187 (2006).
[CrossRef] [PubMed]

Thrall, M. J.

Todo, T.

Y. Kamei, M. Suzuki, K. Watanabe, K. Fujimori, T. Kawasaki, T. Deguchi, Y. Yoneda, T. Todo, S. Takagi, T. Funatsu, and S. Yuba, “Infrared laser-mediated gene induction in targeted single cells in vivo,” Nat. Methods6(1), 79–81 (2009).
[CrossRef] [PubMed]

Wachsmann-Hogiu, S.

S. Wachsmann-Hogiu, T. Weeks, and T. Huser, “Chemical analysis in vivo and in vitro by Raman spectroscopy--from single cells to humans,” Curr. Opin. Biotechnol.20(1), 63–73 (2009).
[CrossRef] [PubMed]

Wada, S.

Wakamatsu, Y.

M. Lynn Lamoreux, R. N. Kelsh, Y. Wakamatsu, and K. Ozato, “Pigment pattern formation in the medaka embryo,” Pigment Cell Res.18(2), 64–73 (2005).
[CrossRef] [PubMed]

Wang, Z.

Watanabe, K.

Y. Kamei, M. Suzuki, K. Watanabe, K. Fujimori, T. Kawasaki, T. Deguchi, Y. Yoneda, T. Todo, S. Takagi, T. Funatsu, and S. Yuba, “Infrared laser-mediated gene induction in targeted single cells in vivo,” Nat. Methods6(1), 79–81 (2009).
[CrossRef] [PubMed]

Weeks, T.

S. Wachsmann-Hogiu, T. Weeks, and T. Huser, “Chemical analysis in vivo and in vitro by Raman spectroscopy--from single cells to humans,” Curr. Opin. Biotechnol.20(1), 63–73 (2009).
[CrossRef] [PubMed]

Wittbrodt, J.

P. J. Keller, A. D. Schmidt, J. Wittbrodt, and E. H. Stelzer, “Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy,” Science322(5904), 1065–1069 (2008).
[CrossRef] [PubMed]

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305(5686), 1007–1009 (2004).
[CrossRef] [PubMed]

Wong, K. K.

Wong, S. T.

Xie, X. S.

B. G. Saar, L. R. Contreras-Rojas, X. S. Xie, and R. H. Guy, “Imaging drug delivery to skin with stimulated Raman scattering microscopy,” Mol. Pharm.8(3), 969–975 (2011).
[CrossRef] [PubMed]

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, “Video-rate molecular imaging in vivo with stimulated Raman scattering,” Science330(6009), 1368–1370 (2010).
[CrossRef] [PubMed]

Yang, Y.

Yoneda, Y.

Y. Kamei, M. Suzuki, K. Watanabe, K. Fujimori, T. Kawasaki, T. Deguchi, Y. Yoneda, T. Todo, S. Takagi, T. Funatsu, and S. Yuba, “Infrared laser-mediated gene induction in targeted single cells in vivo,” Nat. Methods6(1), 79–81 (2009).
[CrossRef] [PubMed]

Yuba, S.

Y. Kamei, M. Suzuki, K. Watanabe, K. Fujimori, T. Kawasaki, T. Deguchi, Y. Yoneda, T. Todo, S. Takagi, T. Funatsu, and S. Yuba, “Infrared laser-mediated gene induction in targeted single cells in vivo,” Nat. Methods6(1), 79–81 (2009).
[CrossRef] [PubMed]

Zhang, Y.

Y. Zhang, H. Hong, and W. Cai, “Imaging with Raman spectroscopy,” Curr. Pharm. Biotechnol.11(6), 654–661 (2010).
[CrossRef] [PubMed]

Zhou, H.

Analyst (Lond.)

C. Krafft, B. Dietzek, and J. Popp, “Raman and CARS microspectroscopy of cells and tissues,” Analyst (Lond.)134(6), 1046–1057 (2009).
[CrossRef] [PubMed]

J. Horsnell, P. Stonelake, J. Christie-Brown, G. Shetty, J. Hutchings, C. Kendall, and N. Stone, “Raman spectroscopy--a new method for the intra-operative assessment of axillary lymph nodes,” Analyst (Lond.)135(12), 3042–3047 (2010).
[CrossRef] [PubMed]

Appl. Spectrosc.

Appl. Spectrosc. Rev.

Z. Movasaghi, S. Rehman, and I. U. Rehman, “Raman spectroscopy of biological tissues,” Appl. Spectrosc. Rev.42(5), 493–541 (2007).
[CrossRef]

Bioessays

J. Huisken, “Slicing embryos gently with laser light sheets,” Bioessays34(5), 406–411 (2012).
[CrossRef] [PubMed]

Biomed. Opt. Express

Clin. Orthop. Relat. Res.

M. D. Morris and G. S. Mandair, “Raman assessment of bone quality,” Clin. Orthop. Relat. Res.469(8), 2160–2169 (2011).
[CrossRef] [PubMed]

Curr. Opin. Biotechnol.

S. Wachsmann-Hogiu, T. Weeks, and T. Huser, “Chemical analysis in vivo and in vitro by Raman spectroscopy--from single cells to humans,” Curr. Opin. Biotechnol.20(1), 63–73 (2009).
[CrossRef] [PubMed]

Curr. Opin. Neurobiol.

P. J. Keller and E. H. Stelzer, “Quantitative in vivo imaging of entire embryos with Digital Scanned Laser Light Sheet Fluorescence Microscopy,” Curr. Opin. Neurobiol.18(6), 624–632 (2008).
[CrossRef] [PubMed]

Curr. Pharm. Biotechnol.

Y. Zhang, H. Hong, and W. Cai, “Imaging with Raman spectroscopy,” Curr. Pharm. Biotechnol.11(6), 654–661 (2010).
[CrossRef] [PubMed]

J. Agric. Food Chem.

S. Thawornchinsombut, J. W. Park, G. Meng, and E. C. Li-Chan, “Raman spectroscopy determines structural changes associated with gelation properties of fish proteins recovered at alkaline pH,” J. Agric. Food Chem.54(6), 2178–2187 (2006).
[CrossRef] [PubMed]

J. Biomed. Opt.

K. Hamada, K. Fujita, N. I. Smith, M. Kobayashi, Y. Inouye, and S. Kawata, “Raman microscopy for dynamic molecular imaging of living cells,” J. Biomed. Opt.13(4), 044027 (2008).
[CrossRef] [PubMed]

J. Raman Spectrosc.

I. Barman, K. M. Tan, and G. P. Singh, “Optical sectioning using single-plane-illumination Raman imaging,” J. Raman Spectrosc.41(10), 1099–1101 (2010).
[CrossRef]

J. Struct. Chem.

G. N. Ten, T. G. Bourova, and V. I. Baranov, “Calculation of Raman spectra and structure elucidation of guanine in the polycrystalline state and in aqueous solution,” J. Struct. Chem.46(6), 998–1005 (2005).
[CrossRef]

Mol. Cells

K. Fujita and N. I. Smith, “Label-free molecular imaging of living cells,” Mol. Cells26(6), 530–535 (2008).
[PubMed]

Mol. Pharm.

B. G. Saar, L. R. Contreras-Rojas, X. S. Xie, and R. H. Guy, “Imaging drug delivery to skin with stimulated Raman scattering microscopy,” Mol. Pharm.8(3), 969–975 (2011).
[CrossRef] [PubMed]

Nat. Chem. Biol.

J. P. Pezacki, J. A. Blake, D. C. Danielson, D. C. Kennedy, R. K. Lyn, and R. Singaravelu, “Chemical contrast for imaging living systems: molecular vibrations drive CARS microscopy,” Nat. Chem. Biol.7(3), 137–145 (2011).
[CrossRef] [PubMed]

Nat. Methods

Y. Kamei, M. Suzuki, K. Watanabe, K. Fujimori, T. Kawasaki, T. Deguchi, Y. Yoneda, T. Todo, S. Takagi, T. Funatsu, and S. Yuba, “Infrared laser-mediated gene induction in targeted single cells in vivo,” Nat. Methods6(1), 79–81 (2009).
[CrossRef] [PubMed]

Opt. Express

Opt. Lett.

Pigment Cell Res.

M. Lynn Lamoreux, R. N. Kelsh, Y. Wakamatsu, and K. Ozato, “Pigment pattern formation in the medaka embryo,” Pigment Cell Res.18(2), 64–73 (2005).
[CrossRef] [PubMed]

Science

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305(5686), 1007–1009 (2004).
[CrossRef] [PubMed]

P. J. Keller, A. D. Schmidt, J. Wittbrodt, and E. H. Stelzer, “Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy,” Science322(5904), 1065–1069 (2008).
[CrossRef] [PubMed]

B. G. Saar, C. W. Freudiger, J. Reichman, C. M. Stanley, G. R. Holtom, and X. S. Xie, “Video-rate molecular imaging in vivo with stimulated Raman scattering,” Science330(6009), 1368–1370 (2010).
[CrossRef] [PubMed]

Other

T. Yamamoto, “Chromatophores and Iridocytes,” MEDAKA(killifish): Biology and Strains, http://www.bio.nagoya-u.ac.jp:8000/iridocytes.html

Supplementary Material (1)

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Figures (6)

Fig. 1
Fig. 1

The light sheet Raman microscopic system. (a) A schematic diagram of the experimental setup of the microscope with the light sheet illumination from the overview and the sideview. The sample was settled at the focal point crossing the illumination laser beam. (b) For lower magnification, the sample was put into a quartz cuvette and was observed with a 5X objective via air. (c) For higher magnification, the sample was settled into the home-made-chamber and was observed by a water immersion lens (20X).

Fig. 2
Fig. 2

(Media 1) Light sheet−excited spontaneous Raman images of a living fish. (a) A dark-field image of a young fish (Quintet line) taken by oblique illumination. (b) Raman image of acetone (fluid) in a cuvette that illuminated by 779 nm of the excitation laser sheet by which Raman signal of acetone can be observed. (c) Raman image of the Quintet taken by the excitation at 740 nm. (d) Merged image of (a) and (c). (e) Raman image of the Quintet taken by the excitation at 770 nm. (f) Merged image of (a) and (d).

Fig. 3
Fig. 3

Image intensity profile of the Raman and autofluorescence signals. The ROIs were defined for Fig. 2(c). Averaged intensities were plotted to the excitation wavelengths to be observed. Lateral axis were indicated with wavenumber unit for Raman shifts and the wavelengths for the excitations.

Fig. 4
Fig. 4

Raman spectra of Quintet medaka. (a) Iridocyte in the eye, (b) muscle in the body trunk, (c) spine, and (d) agarose gel in which the fish was embedded were measured by the commercially-available Raman microscope.

Fig. 5
Fig. 5

Magnified images of the Quintet medaka taken with the 20X water immersion objective. (a) A dark-field image of the body trunk taken by oblique illumination and, (b) the Raman image taken by the excitation at 740 nm. (c) A dark-field image of the eye taken by oblique illumination and, (d) the Raman image taken by the excitation at 745 nm.

Fig. 6
Fig. 6

Image intensity profile of the Raman signals from iridocytes and guanine. Averaged intensities were plotted to the excitation wavelengths to be observed. Lateral axis were indicated with wavenumber unit for Raman shifts and the wavelengths for the excitations. A peak with an asterisk (*) could be stray light.

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