Fluorescence imaging beyond the ballistic regime by ultrasound pulse guided digital phase conjugation

Fluorescence imaging has revolutionized biomedical research over the past three decades. Its high molecular specificity and unrivaled single molecule level sensitivity have enabled breakthroughs in a variety of research fields. For in vivo applications, its major limitation is the superficial imaging depth as random scattering in biological tissues causes exponential attenuation of the ballistic component of a light wave. Here we present fluorescence imaging beyond the ballistic regime by combining single cycle pulsed ultrasound modulation and digital optical phase conjugation. We demonstrate a near isotropic 3D localized sound-light interaction zone. With the exceptionally high optical gain provided by the digital optical phase conjugation system, we can deliver sufficient optical power to a focus inside highly scattering media for not only fluorescence imaging but also a variety of linear and nonlinear spectroscopy measurements. This technology paves the way for many important applications in both fundamental biology research and clinical studies.

[1]  E. Leith,et al.  Holographic Imagery Through Diffusing Media , 1966 .

[2]  Amnon Yariv,et al.  Phase conjugate optics and real time holography (A) , 1978 .

[3]  J. Feinberg,et al.  Phase-conjugating mirror with continuous-wave gain. , 1980, Optics letters.

[4]  Jack Feinberg,et al.  Photorefractive effects and light‐induced charge migration in barium titanate , 1980 .

[5]  W. Denk,et al.  Two-photon imaging to a depth of 1000 microm in living brains by use of a Ti:Al2O3 regenerative amplifier. , 2003, Optics letters.

[6]  F. Del Bene,et al.  Optical Sectioning Deep Inside Live Embryos by Selective Plane Illumination Microscopy , 2004, Science.

[7]  S. Henrickson,et al.  T-cell priming by dendritic cells in lymph nodes occurs in three distinct phases , 2004, Nature.

[8]  J. Lippincott-Schwartz,et al.  Imaging Intracellular Fluorescent Proteins at Nanometer Resolution , 2006, Science.

[9]  Winfried Denk,et al.  On the fundamental imaging-depth limit in two-photon microscopy , 2006 .

[10]  A. Mosk,et al.  Focusing coherent light through opaque strongly scattering media. , 2007, Optics letters.

[11]  G. Lerosey,et al.  Focusing Beyond the Diffraction Limit with Far-Field Time Reversal , 2007, Science.

[12]  A Márquez,et al.  Time fluctuations of the phase modulation in a liquid crystal on silicon display: characterization and effects in diffractive optics. , 2008, Optics express.

[13]  D. Psaltis,et al.  OPTICAL PHASE CONJUGATION FOR TURBIDITY SUPPRESSION IN BIOLOGICAL SAMPLES. , 2008, Nature photonics.

[14]  E. G. van Putten,et al.  Demixing light paths inside disordered metamaterials. , 2008, Optics express.

[15]  A. Mosk,et al.  Universal optimal transmission of light through disordered materials. , 2008, Physical review letters.

[16]  I. Vellekoop.  Controlling the propagation of light in disordered scattering media , 2008, 0807.1087.

[17]  Lihong V. Wang.  Multiscale photoacoustic microscopy and computed tomography. , 2009, Nature photonics.

[18]  Kunal K. Ghosh,et al.  Advances in light microscopy for neuroscience. , 2009, Annual review of neuroscience.

[19]  Changhuei Yang,et al.  Observation of polarization-gate based reconstruction quality improvement during the process of turbidity suppression by optical phase conjugation. , 2009, Applied physics letters.

[20]  Willy Supatto,et al.  Quantitative imaging of collective cell migration during Drosophila gastrulation: multiphoton microscopy and computational analysis , 2009, Nature Protocols.

[21]  Changhuei Yang,et al.  An in vivo study of turbidity suppression by optical phase conjugation (TSOPC) on rabbit ear , 2009, Optics express.

[22]  Changhuei Yang,et al.  Implementation of a digital optical phase conjugation system and its application to study the robustness of turbidity suppression by phase conjugation , 2010, Optics express.

[23]  Demetri Psaltis,et al.  Imaging through turbid layers by scanning the phase conjugated second harmonic radiation from a nanoparticle. , 2010, Optics express.

[24]  M. Davidson,et al.  Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination , 2011, Nature Methods.

[25]  Lihong V. Wang,et al.  Time-reversed ultrasonically encoded optical focusing into scattering media , 2010, Nature photonics.

[26]  O. Katz,et al.  Focusing and compression of ultrashort pulses through scattering media , 2010, 1012.0413.

[27]  Puxiang Lai,et al.  Reflection-mode time-reversed ultrasonically encoded optical focusing into turbid media. , 2011, Journal of biomedical optics.

[28]  Jianyong Tang,et al.  Superpenetration optical microscopy by iterative multiphoton adaptive compensation technique , 2012, Proceedings of the National Academy of Sciences.