Stimulated Raman scattering microscope with shot noise limited sensitivity using subharmonically synchronized laser pulses.

We propose and demonstrate the use of subharmonically synchronized laser pulses for low-noise lock-in detection in stimulated Raman scattering (SRS) microscopy. In the experiment, Yb-fiber laser pulses at a repetition rate of 38 MHz are successfully synchronized to Ti:sapphire laser pulses at a repetition rate of 76 MHz with a jitter of <8 fs by a two-photon detector and an intra-cavity electro-optic modulator. By using these pulses, high-frequency lock-in detection of SRS signal is accomplished without high-speed optical modulation. The noise level of the lock-in signal is found to be higher than the shot noise limit only by 1.6 dB. We also demonstrate high-contrast, 3D imaging of unlabeled living cells.

[1]  K. Itoh,et al.  Analysis and experimental assessment of the sensitivity of stimulated Raman scattering microscopy. , 2009, Optics express.

[2]  Andreas Volkmer,et al.  Vibrational Imaging with High Sensitivity via Epidetected Coherent Anti-Stokes Raman Scattering Microscopy , 2001 .

[3]  Kazuyoshi Itoh,et al.  Stimulated Raman scattering microscopy for live-cell imaging with high contrast and high sensitivity , 2010 .

[4]  C. Shank,et al.  Surface vibrational spectroscopy using stimulated Raman scattering , 1979 .

[5]  Gunay Yurtsever,et al.  Two-color, two-photon, and excited-state absorption microscopy. , 2007, Journal of biomedical optics.

[6]  Jun Ye,et al.  Mode-locked fiber laser frequency-controlled with an intracavity electro-optic modulator. , 2005, Optics letters.

[7]  Satoshi Kawata,et al.  Jitter reduction of two synchronized picosecond mode-locked lasers using balanced cross-correlator with two-photon detectors , 2006 .

[8]  Khanh Kieu,et al.  High-power picosecond fiber source for coherent Raman microscopy. , 2009, Optics letters.

[9]  Jun Ye,et al.  Subfemtosecond timing jitter between two independent, actively synchronized, mode-locked lasers. , 2002, Optics letters.

[10]  X. Xie,et al.  Polarization coherent anti-Stokes Raman scattering microscopy. , 2001, Optics letters.

[11]  Yaron Silberberg,et al.  Nonlinear scanning laser microscopy by third harmonic generation , 1997 .

[12]  Conor L Evans,et al.  Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[13]  T. Murphy,et al.  Broad-band optical clock recovery system using two-photon absorption , 2004, IEEE Photonics Technology Letters.

[14]  Wei Min,et al.  Imaging chromophores with undetectable fluorescence by stimulated emission microscopy , 2009, Nature.

[15]  Feruz Ganikhanov,et al.  Broadly tunable dual-wavelength light source for coherent anti-Stokes Raman scattering microscopy. , 2006, Optics letters.

[16]  W. R. Wiley,et al.  Three-Dimensional Vibrational Imaging by Coherent Anti-Stokes Raman Scattering , 1999 .

[17]  Satoshi Kawata,et al.  Multi-focus excitation coherent anti-Stokes Raman scattering (CARS) microscopy and its applications for real-time imaging. , 2009, Optics express.

[18]  Monika Ritsch-Marte,et al.  Selective imaging of saturated and unsaturated lipids by wide-field CARS-microscopy. , 2008, Optics express.

[19]  S. Kawata,et al.  Molecular vibration imaging in the fingerprint region by use of coherent anti-Stokes Raman scattering microscopy with a collinear configuration. , 2000, Optics letters.

[20]  H. Hamaguchi,et al.  Vibrationally resonant imaging of a single living cell by supercontinuum-based multiplex coherent anti-Stokes Raman scattering microspectroscopy. , 2005, Optics express.

[21]  Jun Ye,et al.  Synchronization of two passively mode-locked, picosecond lasers within 20 fs for coherent anti-Stokes Raman scattering microscopy , 2002 .

[22]  J G Fujimoto,et al.  Attosecond active synchronization of passively mode-locked lasers by balanced cross correlation. , 2003, Optics letters.

[23]  L M Loew,et al.  High-resolution nonlinear optical imaging of live cells by second harmonic generation. , 1999, Biophysical journal.

[24]  Mortazavi,et al.  Supporting Online Material Materials and Methods Figs. S1 to S13 Tables S1 to S3 References Label-free Biomedical Imaging with High Sensitivity by Stimulated Raman Scattering Microscopy , 2022 .

[25]  Conor L Evans,et al.  Coherent anti-stokes raman scattering spectral interferometry: determination of the real and imaginary components of nonlinear susceptibility chi(3) for vibrational microscopy. , 2004, Optics letters.

[26]  James G. Fujimoto,et al.  Attosecond active synchronization of passively mode-locked lasers by balanced cross correlation. , 2003 .

[27]  Feruz Ganikhanov,et al.  High-sensitivity vibrational imaging with frequency modulation coherent anti-Stokes Raman scattering (FM CARS) microscopy. , 2006, Optics letters.

[28]  S. Namiki,et al.  External Synchronization of 160-GHz Optical Beat Signal by Optical Phase-Locked Loop Technique , 2006, IEEE Photonics Technology Letters.

[29]  A. Volkmer,et al.  Vibrational imaging based on stimulated Raman scattering microscopy , 2009 .

[30]  Kazuyoshi Itoh,et al.  Stimulated parametric emission microscopy. , 2006, Optics express.

[31]  Chrisita Ackermann,et al.  Intracavity wavelength modulation of an optical parametric oscillator for coherent Raman microscopy. , 2009, Optics express.