Accurate evaluation of size and refractive index for spherical objects in quantitative phase imaging.

Measuring the average refractive index (RI) of spherical objects, such as suspended cells, in quantitative phase imaging (QPI) requires a decoupling of RI and size from the QPI data. This has been commonly achieved by determining the object's radius with geometrical approaches, neglecting light-scattering. Here, we present a novel QPI fitting algorithm that reliably uncouples the RI using Mie theory and a semi-analytical, corrected Rytov approach. We assess the range of validity of this algorithm in silico and experimentally investigate various objects (oil and protein droplets, microgel beads, cells) and noise conditions. In addition, we provide important practical cues for the analysis of spherical objects in QPI.

[1]  R. Barer,et al.  Refractive Index of Concentrated Protein Solutions , 1954, Nature.

[2]  Jochen Guck,et al.  Quantifying cellular differentiation by physical phenotype using digital holographic microscopy. , 2012, Integrative biology : quantitative biosciences from nano to macro.

[3]  Gene W. Yeo,et al.  Misregulated RNA processing in amyotrophic lateral sclerosis , 2012, Brain Research.

[4]  Christian Depeursinge,et al.  Determination of Transmembrane Water Fluxes in Neurons Elicited by Glutamate Ionotropic Receptors and by the Cotransporters KCC2 and NKCC1: A Digital Holographic Microscopy Study , 2011, The Journal of Neuroscience.

[5]  Paul Müller,et al.  ODTbrain: a Python library for full-view, dense diffraction tomography , 2015, BMC Bioinformatics.

[6]  Jindong Tian,et al.  Quantitative refractive index distribution of single cell by combining phase-shifting interferometry and AFM imaging , 2017, Scientific Reports.

[7]  Christian Depeursinge,et al.  Cell morphology and intracellular ionic homeostasis explored with a multimodal approach combining epifluorescence and digital holographic microscopy , 2010, Journal of biophotonics.

[8]  A. Devaney Inverse-scattering theory within the Rytov approximation. , 1981, Optics letters.

[9]  Ya-Wei Wang,et al.  Relation between substructure position of phase objects in optical axial direction and phase information in quantitative phase imaging , 2014 .

[10]  Jochen Guck,et al.  Three‐dimensional correlative single‐cell imaging utilizing fluorescence and refractive index tomography , 2017, Journal of biophotonics.

[11]  J. Käs,et al.  The optical stretcher: a novel laser tool to micromanipulate cells. , 2001, Biophysical journal.

[12]  J. Guck,et al.  Cell nuclei have lower refractive index and mass density than cytoplasm , 2016, Journal of biophotonics.

[13]  I.‐Yin Sandy Lee,et al.  Calculation of the Mie scattering field inside and outside a coated spherical particle , 2009 .

[14]  Marco Y. Hein,et al.  A Liquid-to-Solid Phase Transition of the ALS Protein FUS Accelerated by Disease Mutation , 2015, Cell.

[15]  Jochen Guck,et al.  Refractive index measurements of single, spherical cells using digital holographic microscopy. , 2018, Methods in cell biology.

[16]  Li-Huei Tsai,et al.  Interaction of FUS and HDAC1 regulates DNA damage response and repair in neurons , 2013, Nature Neuroscience.

[17]  J. Guck,et al.  The Theory of Diffraction Tomography , 2015, 1507.00466.

[18]  Daniel Carl,et al.  Investigation of living pancreas tumor cells by digital holographic microscopy. , 2006, Journal of biomedical optics.

[19]  Vladislav V. Yakovlev,et al.  Seeing cells in a new light: a renaissance of Brillouin spectroscopy , 2016 .

[20]  L. E. Larsen,et al.  Limitations of Imaging with First-Order Diffraction Tomography , 1984 .

[21]  R. Barer Interference Microscopy and Mass Determination , 1952, Nature.

[22]  Patrik Langehanenberg,et al.  Integral refractive index determination of living suspension cells by multifocus digital holographic phase contrast microscopy. , 2007, Journal of biomedical optics.

[23]  Natan T. Shaked,et al.  Whole-cell-analysis of live cardiomyocytes using wide-field interferometric phase microscopy , 2010, Biomedical optics express.

[24]  B. Chen,et al.  Validity of diffraction tomography based on the first born and the first rytov approximations. , 1998, Applied optics.

[25]  William J. Polacheck,et al.  Noncontact three-dimensional mapping of intracellular hydro-mechanical properties by Brillouin microscopy , 2015, Nature Methods.

[26]  T. Klar,et al.  Shaping emission spectra of fluorescent molecules with single plasmonic nanoresonators. , 2008, Physical review letters.

[27]  B. Kemper,et al.  Digital Holographic Microscopy for Quantitative Live Cell Imaging and Cytometry , 2011 .

[28]  J. Guck,et al.  Near- and far-field scattering from arbitrary three-dimensional aggregates of coated spheres using parallel computing. , 2011, Physical review. E, Statistical, nonlinear, and soft matter physics.

[29]  Koichi Iwata,et al.  Calculation of Refractive Index Distribution from Interferograms Using the Born and Rytov's Approximation , 1975 .

[30]  Faramarz Farahi,et al.  Hybrid shear force feedback/scanning quantitative phase microscopy applied to subsurface imaging. , 2009, Optics express.

[31]  Jochen Guck,et al.  Bacterial infection of macrophages induces decrease in refractive index , 2013, Journal of biophotonics.

[32]  C. Fang-Yen,et al.  Optical diffraction tomography for high resolution live cell imaging. , 2009, Optics express.

[33]  Jochen Guck,et al.  Comparison of stresses on homogeneous spheroids in the optical stretcher computed with geometrical optics and generalized Lorenz-Mie theory. , 2012, Applied optics.

[34]  K. Nugent,et al.  Refractive index measurement in viable cells using quantitative phase‐amplitude microscopy and confocal microscopy , 2005, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[35]  Markus Fratz,et al.  Noninvasive time-dependent cytometry monitoring by digital holography. , 2007, Journal of biomedical optics.

[36]  Yongkeun Park,et al.  Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum , 2008, Proceedings of the National Academy of Sciences.

[37]  Avinash C. Kak,et al.  Principles of computerized tomographic imaging , 2001, Classics in applied mathematics.

[38]  K. Badizadegan,et al.  Live cell refractometry using microfluidic devices. , 2006, Optics letters.

[39]  S. Anna,et al.  Microfluidic methods for generating continuous droplet streams , 2007 .

[40]  E. Wolf Three-dimensional structure determination of semi-transparent objects from holographic data , 1969 .

[41]  E. Cuche,et al.  Measurement of the integral refractive index and dynamic cell morphometry of living cells with digital holographic microscopy. , 2005, Optics express.

[42]  B. Wattellier,et al.  Quadriwave lateral shearing interferometry for quantitative phase microscopy of living cells. , 2009, Optics express.

[43]  Christian Depeursinge,et al.  Simultaneous cell morphometry and refractive index measurement with dual-wavelength digital holographic microscopy and dye-enhanced dispersion of perfusion medium. , 2008, Optics letters.

[44]  Adam Wax,et al.  Is the nuclear refractive index lower than cytoplasm? Validation of phase measurements and implications for light scattering technologies , 2017, Journal of biophotonics.