Abstract

Multi-photon microscopy is a powerful tool in biomolecular research. Less complex and more cost effective excitation light sources will make this technique accessible to a broader community. Semiconductor diode seeded fiber lasers have proven to be especially robust, low cost and easy to use. However, their wavelength tuning range is often limited, so only a limited number of fluorophores can be accessed. Therefore, different approaches have been proposed to extend the spectral coverage of these lasers. Recently, we showed that four-wave mixing (FWM) assisted stimulated Raman scattering (SRS) can be harnessed to red-shift high power pulses from 1064 nm to a narrowband output at 1122 nm and 1186 nm and therefore extend the number of accessible fluorophores. In this contribution, we show the applicability of all three wavelengths for multi-photon microscopy and analyze the performance.

© 2018 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

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References

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    [Crossref] [PubMed]

2018 (1)

2017 (6)

W. Fu, L. G. Wright, and F. W. Wise, “High-power femtosecond pulses without a modelocked laser,” Optica 4(7), 831–834 (2017).
[Crossref] [PubMed]

P. Cadroas, L. Abdeladim, L. Kotov, M. Likhachev, D. Lipatov, D. Gaponov, A. Hideur, M. Tang, J. Livet, W. Supatto, E. Beaurepaire, and S. Février, “All-fiber femtosecond laser providing 9 nJ, 50 MHz pulses at 1650 nm for three-photon microscopy,” J. Opt. 19(6), 065506 (2017).
[Crossref]

M. Eibl, S. Karpf, H. Hakert, T. Blömker, J. P. Kolb, C. Jirauschek, and R. Huber, “Pulse-to-pulse wavelength switching of a nanosecond fiber laser by four-wave mixing seeded stimulated Raman amplification,” Opt. Lett. 42(21), 4406–4409 (2017).
[Crossref] [PubMed]

V. R. Supradeepa, F. Yan, and W. N. Jeffrey, “Raman fiber lasers,” J. Opt. 19(2), 023001 (2017).
[Crossref]

H. Hakert, M. Eibl, S. Karpf, and R. Huber, “Sparse-sampling with time-encoded (TICO) stimulated Raman scattering for fast image acquisition,” Proc. SPIE 10414, 1041408 (2017).
[Crossref]

M. Eibl, S. Karpf, D. Weng, H. Hakert, T. Pfeiffer, J. P. Kolb, and R. Huber, “Single pulse two photon fluorescence lifetime imaging (SP-FLIM) with MHz pixel rate,” Biomed. Opt. Express 8(7), 3132–3142 (2017).
[Crossref] [PubMed]

2016 (2)

C. Lefort, R. P. O’Connor, V. Blanquet, L. Magnol, H. Kano, V. Tombelaine, P. Lévêque, V. Couderc, and P. Leproux, “Multicolor multiphoton microscopy based on a nanosecond supercontinuum laser source,” J. Biophotonics 9(7), 709–714 (2016).
[Crossref] [PubMed]

S. Karpf, M. Eibl, B. Sauer, F. Reinholz, G. Hüttmann, and R. Huber, “Two-photon microscopy using fiber-based nanosecond excitation,” Biomed. Opt. Express 7(7), 2432–2440 (2016).
[Crossref] [PubMed]

2015 (10)

T. Gottschall, T. Meyer, M. Schmitt, J. Popp, J. Limpert, and A. Tünnermann, “Four-wave-mixing-based optical parametric oscillator delivering energetic, tunable, chirped femtosecond pulses for non-linear biomedical applications,” Opt. Express 23(18), 23968–23977 (2015).
[Crossref] [PubMed]

R. Kawakami, K. Sawada, Y. Kusama, Y.-C. Fang, S. Kanazawa, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, “In vivo two-photon imaging of mouse hippocampal neurons in dentate gyrus using a light source based on a high-peak power gain-switched laser diode,” Biomed. Opt. Express 6(3), 891–901 (2015).
[Crossref] [PubMed]

T. H. Runcorn, R. T. Murray, E. J. R. Kelleher, S. V. Popov, and J. R. Taylor, “Duration-tunable picosecond source at 560 nm with watt-level average power,” Opt. Lett. 40(13), 3085–3088 (2015).
[Crossref] [PubMed]

H. Zhang, R. Tao, P. Zhou, X. Wang, and X. Xu, “1.5-kW Yb-Raman Combined Nonlinear Fiber Amplifier at 1120 nm,” IEEE Photonics Technol. Lett. 27(6), 628–630 (2015).
[Crossref]

S. Karpf, M. Eibl, W. Wieser, T. Klein, and R. Huber, “A Time-Encoded Technique for fibre-based hyperspectral broadband stimulated Raman microscopy,” Nat. Commun. 6(1), 6784 (2015).
[Crossref] [PubMed]

J. M. Mayrhofer, F. Haiss, D. Haenni, S. Weber, M. Zuend, M. J. P. Barrett, K. D. Ferrari, P. Maechler, A. S. Saab, J. L. Stobart, M. T. Wyss, H. Johannssen, H. Osswald, L. M. Palmer, V. Revol, C.-D. Schuh, C. Urban, A. Hall, M. E. Larkum, E. Rutz-Innerhofer, H. U. Zeilhofer, U. Ziegler, and B. Weber, “Design and performance of an ultra-flexible two-photon microscope for in vivo research,” Biomed. Opt. Express 6(11), 4228–4237 (2015).
[Crossref] [PubMed]

J. P. Kolb, T. Klein, C. L. Kufner, W. Wieser, A. S. Neubauer, and R. Huber, “Ultra-widefield retinal MHz-OCT imaging with up to 100 degrees viewing angle,” Biomed. Opt. Express 6(5), 1534–1552 (2015).
[Crossref] [PubMed]

T. H. Runcorn, T. Legg, R. T. Murray, E. J. R. Kelleher, S. V. Popov, and J. R. Taylor, “Fiber-integrated frequency-doubling of a picosecond Raman laser to 560 nm,” Opt. Express 23(12), 15728–15733 (2015).
[Crossref] [PubMed]

M. Inoue, A. Takeuchi, S. Horigane, M. Ohkura, K. Gengyo-Ando, H. Fujii, S. Kamijo, S. Takemoto-Kimura, M. Kano, J. Nakai, K. Kitamura, and H. Bito, “Rational design of a high-affinity, fast, red calcium indicator R-CaMP2.,” Nat. Methods 12(1), 64–70 (2015).
[Crossref] [PubMed]

C. Tischbirek, A. Birkner, H. Jia, B. Sakmann, and A. Konnerth, “Deep two-photon brain imaging with a red-shifted fluorometric Ca2+ indicator,” Proc. Natl. Acad. Sci. U.S.A. 112(36), 11377–11382 (2015).
[Crossref] [PubMed]

2014 (1)

2013 (3)

E. E. Hoover and J. A. Squier, “Advances in multiphoton microscopy technology,” Nat. Photonics 7(2), 93–101 (2013).
[Crossref] [PubMed]

R. Kawakami, K. Sawada, A. Sato, T. Hibi, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, “Visualizing hippocampal neurons with in vivo two-photon microscopy using a 1030 nm picosecond pulse laser,” Sci. Rep. 3(1), 1014 (2013).
[Crossref] [PubMed]

C. Xu and F. W. Wise, “Recent advances in fibre lasers for nonlinear microscopy,” Nat. Photonics 7(11), 875–882 (2013).
[Crossref] [PubMed]

2012 (1)

2011 (3)

T. Klein, W. Wieser, C. M. Eigenwillig, B. R. Biedermann, and R. Huber, “Megahertz OCT for ultrawide-field retinal imaging with a 1050 nm Fourier domain mode-locked laser,” Opt. Express 19(4), 3044–3062 (2011).
[Crossref] [PubMed]

M. Drobizhev, N. S. Makarov, S. E. Tillo, T. E. Hughes, and A. Rebane, “Two-photon absorption properties of fluorescent proteins,” Nat. Methods 8(5), 393–399 (2011).
[Crossref] [PubMed]

D. Kobat, N. G. Horton, and C. Xu, “In vivo two-photon microscopy to 1.6-mm depth in mouse cortex,” J. Biomed. Opt. 16(10), 106014 (2011).
[Crossref] [PubMed]

2009 (1)

2008 (2)

D. Träutlein, F. Adler, K. Moutzouris, A. Jeromin, A. Leitenstorfer, and E. Ferrando-May, “Highly versatile confocal microscopy system based on a tunable femtosecond Er:fiber source,” J. Biophotonics 1(1), 53–61 (2008).
[Crossref] [PubMed]

N. S. Makarov, M. Drobizhev, and A. Rebane, “Two-photon absorption standards in the 550-1600 nm excitation wavelength range,” Opt. Express 16(6), 4029–4047 (2008).
[Crossref] [PubMed]

2007 (1)

M. Drobizhev, N. S. Makarov, T. Hughes, and A. Rebane, “Resonance Enhancement of Two-Photon Absorption in Fluorescent Proteins,” J. Phys. Chem. B 111(50), 14051–14054 (2007).
[Crossref] [PubMed]

2006 (1)

2005 (1)

F. Helmchen and W. Denk, “Deep tissue two-photon microscopy,” Nat. Methods 2(12), 932–940 (2005).
[Crossref] [PubMed]

2003 (1)

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[Crossref] [PubMed]

2000 (1)

P. T. C. So, C. Y. Dong, B. R. Masters, and K. M. Berland, “Two-Photon Excitation Fluorescence Microscopy,” Annu. Rev. Biomed. Eng. 2(1), 399–429 (2000).
[Crossref] [PubMed]

1998 (1)

Bewersdorf and Hell, “Picosecond pulsed two-photon imaging with repetition rates of 200 and 400 MHz,” J. Microsc. 191(1), 28–38 (1998).
[Crossref]

1991 (1)

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and et, “Optical coherence tomography,” Science 254(5035), 1178–1181 (1991).
[Crossref] [PubMed]

1990 (1)

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248(4951), 73–76 (1990).
[Crossref] [PubMed]

1973 (1)

R. H. Stolen and E. P. Ippen, “Raman gain in glass optical waveguides,” Appl. Phys. Lett. 22(6), 276–278 (1973).
[Crossref]

Abdeladim, L.

P. Cadroas, L. Abdeladim, L. Kotov, M. Likhachev, D. Lipatov, D. Gaponov, A. Hideur, M. Tang, J. Livet, W. Supatto, E. Beaurepaire, and S. Février, “All-fiber femtosecond laser providing 9 nJ, 50 MHz pulses at 1650 nm for three-photon microscopy,” J. Opt. 19(6), 065506 (2017).
[Crossref]

Adler, F.

D. Träutlein, F. Adler, K. Moutzouris, A. Jeromin, A. Leitenstorfer, and E. Ferrando-May, “Highly versatile confocal microscopy system based on a tunable femtosecond Er:fiber source,” J. Biophotonics 1(1), 53–61 (2008).
[Crossref] [PubMed]

Barrett, M. J. P.

Beaurepaire, E.

P. Cadroas, L. Abdeladim, L. Kotov, M. Likhachev, D. Lipatov, D. Gaponov, A. Hideur, M. Tang, J. Livet, W. Supatto, E. Beaurepaire, and S. Février, “All-fiber femtosecond laser providing 9 nJ, 50 MHz pulses at 1650 nm for three-photon microscopy,” J. Opt. 19(6), 065506 (2017).
[Crossref]

Berland, K. M.

P. T. C. So, C. Y. Dong, B. R. Masters, and K. M. Berland, “Two-Photon Excitation Fluorescence Microscopy,” Annu. Rev. Biomed. Eng. 2(1), 399–429 (2000).
[Crossref] [PubMed]

Bewersdorf,

Bewersdorf and Hell, “Picosecond pulsed two-photon imaging with repetition rates of 200 and 400 MHz,” J. Microsc. 191(1), 28–38 (1998).
[Crossref]

Biedermann, B. R.

Birkner, A.

C. Tischbirek, A. Birkner, H. Jia, B. Sakmann, and A. Konnerth, “Deep two-photon brain imaging with a red-shifted fluorometric Ca2+ indicator,” Proc. Natl. Acad. Sci. U.S.A. 112(36), 11377–11382 (2015).
[Crossref] [PubMed]

Bito, H.

M. Inoue, A. Takeuchi, S. Horigane, M. Ohkura, K. Gengyo-Ando, H. Fujii, S. Kamijo, S. Takemoto-Kimura, M. Kano, J. Nakai, K. Kitamura, and H. Bito, “Rational design of a high-affinity, fast, red calcium indicator R-CaMP2.,” Nat. Methods 12(1), 64–70 (2015).
[Crossref] [PubMed]

Blanquet, V.

C. Lefort, R. P. O’Connor, V. Blanquet, L. Magnol, H. Kano, V. Tombelaine, P. Lévêque, V. Couderc, and P. Leproux, “Multicolor multiphoton microscopy based on a nanosecond supercontinuum laser source,” J. Biophotonics 9(7), 709–714 (2016).
[Crossref] [PubMed]

Blömker, T.

Cadroas, P.

P. Cadroas, L. Abdeladim, L. Kotov, M. Likhachev, D. Lipatov, D. Gaponov, A. Hideur, M. Tang, J. Livet, W. Supatto, E. Beaurepaire, and S. Février, “All-fiber femtosecond laser providing 9 nJ, 50 MHz pulses at 1650 nm for three-photon microscopy,” J. Opt. 19(6), 065506 (2017).
[Crossref]

Calia, D. B.

Chan, M.-C.

Chang, W.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and et, “Optical coherence tomography,” Science 254(5035), 1178–1181 (1991).
[Crossref] [PubMed]

Couderc, V.

C. Lefort, R. P. O’Connor, V. Blanquet, L. Magnol, H. Kano, V. Tombelaine, P. Lévêque, V. Couderc, and P. Leproux, “Multicolor multiphoton microscopy based on a nanosecond supercontinuum laser source,” J. Biophotonics 9(7), 709–714 (2016).
[Crossref] [PubMed]

Denk, W.

F. Helmchen and W. Denk, “Deep tissue two-photon microscopy,” Nat. Methods 2(12), 932–940 (2005).
[Crossref] [PubMed]

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248(4951), 73–76 (1990).
[Crossref] [PubMed]

Dong, C. Y.

P. T. C. So, C. Y. Dong, B. R. Masters, and K. M. Berland, “Two-Photon Excitation Fluorescence Microscopy,” Annu. Rev. Biomed. Eng. 2(1), 399–429 (2000).
[Crossref] [PubMed]

Drobizhev, M.

M. Drobizhev, N. S. Makarov, S. E. Tillo, T. E. Hughes, and A. Rebane, “Two-photon absorption properties of fluorescent proteins,” Nat. Methods 8(5), 393–399 (2011).
[Crossref] [PubMed]

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Optica (1)

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

C. Tischbirek, A. Birkner, H. Jia, B. Sakmann, and A. Konnerth, “Deep two-photon brain imaging with a red-shifted fluorometric Ca2+ indicator,” Proc. Natl. Acad. Sci. U.S.A. 112(36), 11377–11382 (2015).
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Proc. SPIE (1)

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Figures (6)

Fig. 1
Fig. 1 Setup of the multi-color master oscillator power amplifier (MOPA). The modulated output of a 1064 nm seed laser diode is amplified in a multistage ytterbium doped fiber amplifier (YDFA) to ~1 kW peak power. These high power pulses are shifted to longer wavelengths by a combination of four-wave mixing (FWM) and stimulated Raman scattering (SRS) in passive standard single mode fiber. By controlling the pump power and the power of the 1122 nm Raman seed laser, the operation can be switched between three wavelengths [27]. The fiber output makes it easy to couple the laser to any multi-photon microscope. EOM: electro optical modulator, Yb: Ytterbium, WDM: wavelength division multiplexer, C: collimator, GM: galvanometric mirrors, SL: scan lens, TL: tube lens, CL: condenser lens, DM: dichroic mirror, PMT: photo-multiplier tube.
Fig. 2
Fig. 2 Emission spectra of the multi-color laser. a) – c) The output spectra when the laser emission is optimized for 1064 nm, 1122 nm, and 1186 nm, respectively. This conversion is the result of the FWM-assisted SRS effect [27]. The Raman gain curve, reproduced to scale after Stolen and Ippen [28], is indicated by grey areas. The spectral components are equidistant in energy with a difference of 14.4 THz as expected from a FWM process.
Fig. 3
Fig. 3 Electronic synchronization of the homebuilt multi-photon microscope. An arbitrary waveform generator (AWG) provides a sample clock (CLK) reference signal and trigger signals for all components. ADC: analog-to-digital converter card, PMT: photomultiplier tube, GM: galvanometric mirrors.
Fig. 4
Fig. 4 One-photon absorption and emission spectra for the dyes Atto-594 and Star-635 which are used to label the mitochondria and tubulin in the COS-7, respectively. Solid curves are absorption spectra, dashed curves emission spectra. Colored lines indicate half values of the available excitation wavelengths.
Fig. 5
Fig. 5 Imaging with 1064 nm excitation light. a) TPEF image of a stained slice of convalaria majalis. b) TPEF auto fluorescence image of a ficus benjamina plant leave. c) SHG image of mouse skin. Scale bar 100 µm.
Fig. 6
Fig. 6 Two-photon imaging of stained COS-7 cells with long wavelength excitation. a) 1122 nm excitation channel. b) 1186 nm excitation channel. Images c) and d) are the color-coded fluorochrome channels, where d) is a result of subtracting a) from b) to isolate the Atto-594 channel. e) Processed overlay, where green color shows mitochondria (Atto-594) and blue color tubulin (STAR-635). Scale bar 50 µm.

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