Phase-Retrieved Tomography enables Mesoscopic imaging of Opaque Tumor Spheroids

We present a new Phase-Retrieved Tomography (PRT) method to radically improve mesoscopic imaging at regimes beyond one transport mean-free-path and achieve high resolution, uniformly throughout the volume of opaque samples. The method exploits multi-view acquisition in a hybrid Selective Plane Illumination Microscope (SPIM) and Optical Projection Tomography (OPT) setup and a three-dimensional Gerchberg-Saxton phase-retrieval algorithm applied in 3D through the autocorrelation sinogram. We have successfully applied this innovative protocol to image optically dense 3D cell cultures in the form of tumor spheroids, highly versatile models to study cancer behavior and response to chemotherapy. We have thus achieved a significant improvement of resolution in depths not yet accessible with the currently used methods in SPIM/OPT, while overcoming all registration and alignment problems inherent to these techniques.

[1]  C. Felser,et al.  Negative magnetoresistance without well-defined chirality in the Weyl semimetal TaP , 2015, Nature Communications.

[2]  Chandra Sekhar Seelamantula,et al.  An iterative algorithm for phase retrieval with sparsity constraints: application to frequency domain optical coherence tomography , 2012, 2012 IEEE International Conference on Acoustics, Speech and Signal Processing (ICASSP).

[3]  S. Hell.  Far-Field Optical Nanoscopy , 2007, Science.

[4]  J. Miao,et al.  Oversampling smoothness: an effective algorithm for phase retrieval of noisy diffraction intensities. , 2012, Journal of applied crystallography.

[5]  M. Fink,et al.  Non-invasive single-shot imaging through scattering layers and around corners via speckle correlations , 2014, Nature Photonics.

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

[7]  J. Huisken,et al.  Omnidirectional microscopy , 2012, Nature Methods.

[8]  J. Hecksher-Sørensen,et al.  Optical Projection Tomography as a Tool for 3D Microscopy and Gene Expression Studies , 2002, Science.

[9]  Daniele Ancora,et al.  Optical projection tomography via phase retrieval algorithms for hidden three dimensional imaging , 2017, BiOS.

[10]  Jan Huisken,et al.  Selective plane illumination microscopy techniques in developmental biology , 2009, Development.

[11]  Vasilis Ntziachristos,et al.  In vivo imaging of Drosophila melanogaster pupae with mesoscopic fluorescence tomography , 2007, Nature Methods.

[12]  Sylvain Gigan,et al.  Optical microscopy aims deep , 2017, Nature Photonics.

[13]  Jorge Ripoll,et al.  Microscopic Optical Projection Tomography In Vivo , 2011, PloS one.

[14]  Jie Tian,et al.  Helical optical projection tomography. , 2013, Optics express.

[15]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[16]  Benjamin Schmid,et al.  High-resolution reconstruction of the beating zebrafish heart , 2014, Nature Methods.

[17]  D. Stainier,et al.  Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM). , 2007, Optics letters.

[18]  W. Webb,et al.  Nonlinear magic: multiphoton microscopy in the biosciences , 2003, Nature Biotechnology.

[19]  Feng,et al.  Memory effects in propagation of optical waves through disordered media. , 1988, Physical review letters.

[20]  Jorge Ripoll,et al.  Correction for specimen movement and rotation errors for in-vivo Optical Projection Tomography , 2010, Biomedical optics express.

[21]  G. Lerosey,et al.  Controlling waves in space and time for imaging and focusing in complex media , 2012, Nature Photonics.

[22]  Xavier Intes,et al.  Mesoscopic Fluorescence Molecular Tomography for Evaluating Engineered Tissues , 2015, Annals of Biomedical Engineering.

[23]  James Sharpe,et al.  Tomographic molecular imaging and 3D quantification within adult mouse organs , 2007, Nature Methods.

[24]  Daniele Ancora,et al.  Phase-retrieved optical projection tomography for 3D imaging through scattering layers , 2016, SPIE BiOS.

[25]  L. Kunz-Schughart,et al.  Multicellular tumor spheroids: an underestimated tool is catching up again. , 2010, Journal of biotechnology.

[26]  V. Ntziachristos.  Going deeper than microscopy: the optical imaging frontier in biology , 2010, Nature Methods.

[27]  Juergen Friedrich,et al.  Spheroid-based drug screen: considerations and practical approach , 2009, Nature Protocols.

[28]  Fredrik Georgsson,et al.  Image Processing Assisted Algorithms for Optical Projection Tomography , 2012, IEEE Transactions on Medical Imaging.

[29]  Victoria J Allan,et al.  Light Microscopy Techniques for Live Cell Imaging , 2003, Science.

[30]  J. Bertolotti,et al.  Non-invasive imaging through opaque scattering layers , 2012, Nature.

[31]  P. Verveer,et al.  High-resolution three-dimensional imaging of large specimens with light sheet–based microscopy , 2007, Nature Methods.

[32]  V. Ntziachristos,et al.  FMT-XCT: in vivo animal studies with hybrid fluorescence molecular tomography–X-ray computed tomography , 2012, Nature Methods.

[33]  Stephan Preibisch,et al.  Efficient Bayesian-based multiview deconvolution , 2013, Nature Methods.

[34]  Daniele Ancora,et al.  Tailored light sheets through opaque cylindrical lenses , 2016 .

[35]  Jan Huisken,et al.  Multi-view image fusion improves resolution in three-dimensional microscopy. , 2007, Optics express.

[36]  Daniele Ancora,et al.  Tailoring non-diffractive beams from amorphous light speckles , 2016 .

[37]  Lars Hufnagel,et al.  Multiview light-sheet microscope for rapid in toto imaging , 2012, Nature Methods.

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

[39]  Liang Gao,et al.  Imaging multicellular specimens with real-time optimized tiling light-sheet selective plane illumination microscopy , 2016, Nature Communications.

[40]  Sun Qiaochu,et al.  Application to Frequency Domain Optical Coherence Tomography (OCT) to Dental composite resin restored tooth , 2016 .

[41]  Daniel Kolditz,et al.  Iterative reconstruction methods in X-ray CT. , 2012, Physica medica : PM : an international journal devoted to the applications of physics to medicine and biology : official journal of the Italian Association of Biomedical Physics.

[42]  Yonina C. Eldar,et al.  Phase Retrieval with Application to Optical Imaging: A contemporary overview , 2015, IEEE Signal Processing Magazine.

[43]  James R Fienup,et al.  Phase retrieval algorithms: a personal tour [Invited]. , 2013, Applied optics.

[44]  Yonina C. Eldar,et al.  Phase Retrieval with Application to Optical Imaging , 2014, ArXiv.

[45]  Jeff W. Lichtman,et al.  Clarifying Tissue Clearing , 2015, Cell.

[46]  Jorge Ripoll,et al.  A Customized Light Sheet Microscope to Measure Spatio-Temporal Protein Dynamics in Small Model Organisms , 2015, PloS one.

[47]  Simon Arridge,et al.  Accelerated Optical Projection Tomography Applied to In Vivo Imaging of Zebrafish , 2015, PloS one.

[48]  Jie Tian,et al.  Automated Recovery of the Center of Rotation in Optical Projection Tomography in the Presence of Scattering , 2013, IEEE Journal of Biomedical and Health Informatics.