Optical diffraction tomography for high resolution live cell imaging Citation

We report the experimental implementation of optical diffraction tomography for quantitative 3D mapping of refractive index in live biological cells. Using a heterodyne Mach-Zehnder interferometer, we record complex field images of light transmitted through a sample with varying directions of illumination. To quantitatively reconstruct the 3D map of complex refractive index in live cells, we apply optical diffraction tomography based on the Rytov approximation. In this way, the effect of diffraction is taken into account in the reconstruction process and diffraction-free high resolution 3D images are obtained throughout the entire sample volume. The quantitative refractive index map can potentially serve as an intrinsic assay to provide the molecular concentrations without the addition of exogenous agents and also to provide a method for studying the light scattering properties of single cells. ©2008 Optical Society of America OCIS codes: (120.3180) Interferometry; (180.0180) Microscopy; (170.3880) Medical and biological imaging. References and links 1. F. Zernike, "Phase-contrast, a new method for microscopic observation of transparent objects. Part I.," Physica 9, 686-698 (1942). 2. G. Nomarski, "Microinterféromètre différentiel à ondes polarisées," J. Phys. Radium 16, 9S-11S (1955). 3. Y. K. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, "Diffraction phase and fluorescence microscopy," Opt. Express 14, 8263-8268 (2006). 4. G. Popescu, T. Ikeda, R. R. Dasari, and M. S. Feld, "Diffraction phase microscopy for quantifying cell structure and dynamics," Opt Lett 31, 775-777 (2006). 5. C. Fang-Yen, S. Oh, Y. Park, W. Choi, S. Song, H. S. Seung, R. R. Dasari, and M. S. Feld, "Imaging voltage-dependent cell motions with heterodyne Mach-Zehnder phase microscopy," Opt Lett 32, 1572-1574 (2007). 6. P. Marquet, B. Rappaz, P. J. Magistretti, E. Cuche, Y. Emery, T. Colomb, and C. Depeursinge, "Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy," Opt Lett 30, 468-470 (2005). 7. A. Barty, K. A. Nugent, D. Paganin, and A. Roberts, "Quantitative optical phase microscopy," Opt Lett 23, 817-819 (1998). 8. G. Popescu, T. Ikeda, K. Goda, C. A. Best-Popescu, M. Laposata, S. Manley, R. R. Dasari, K. Badizadegan, and M. S. Feld, "Optical measurement of cell membrane tension," Phys Rev Lett 97, 218101 (2006). 9. B. Rappaz, P. Marquet, E. Cuche, Y. Emery, C. Depeursinge, and P. Magistretti, "Measurement of the integral refractive index and dynamic cell morphometry of living cells with digital holographic microscopy," Opt. Express 13, 9361-9373 (2005). 10. N. Lue, G. Popescu, T. Ikeda, R. R. Dasari, K. Badizadegan, and M. S. Feld, "Live cell refractometry using microfluidic devices," Opt. Lett. 31, 2759-2761 (2006). 11. B. Rappaz, F. Charriere, C. Depeursinge, P. J. Magistretti, and P. Marquet, "Simultaneous cell morphometry and refractive index measurement with dual-wavelength digital holographic microscopy and dye-enhanced dispersion of perfusion medium," Opt Lett 33, 744-746 (2008). 12. A. Barty, K. A. Nugent, A. Roberts, and D. Paganin, "Quantitative phase tomography," Optics Communications 175, 329-336 (2000). 13. W. Gorski, and W. Osten, "Tomographic imaging of photonic crystal fibers," Opt Lett 32, 1977-1979 (2007). 14. F. Charriere, N. Pavillon, T. Colomb, C. Depeursinge, T. J. Heger, E. A. D. Mitchell, P. Marquet, and B. Rappaz, "Living specimen tomography by digital holographic microscopy: morphometry of testate amoeba," Opt Express 14, 7005-7013 (2006). 15. V. Lauer, "New approach to optical diffraction tomography yielding a vector equation of diffraction tomography and a novel tomographic microscope," J. Microsc. 205, 165-176 (2002). 16. W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, "Tomographic phase microscopy," Nature Methods 4, 717-719 (2007). 17. K. C. Tam, and V. Perezmendez, "Tomographical Imaging with Limited-Angle Input," J Opt Soc Am 71, 582-592 (1981). 18. B. P. Medoff, W. R. Brody, M. Nassi, and A. Macovski, "Iterative Convolution Backprojection Algorithms for Image-Reconstruction from Limited Data," J. Opt. Soc. Am. 73, 1493-1500 (1983). 19. A. C. Kak, and M. Slaney, Principles of Computerized Tomographic Imaging (Academic Press, New York, 1999). 20. E. Wolf, "Three-dimensional structure determination of semi-transparent objects from holographic data," Opt. Commun. 1, 153-156 (1969). 21. M. Debailleul, B. Simon, V. Georges, O. Haeberle, and V. Lauer, "Holographic microscopy and diffractive microtomography of transparent samples," Meas Sci Technol 19, (2008). 22. W. S. Choi, C. Fang-Yen, K. Badizadegan, R. R. Dasari, and M. S. Feld, "Extended depth of focus in tomographic phase microscopy using a propagation algorithm," Opt Lett 33, 171-173 (2008). 23. Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, and S. Suresh, "Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum," Proc Natl Acad Sci U S A 105, 13730-13735 (2008). 24. W. Choi, C. C. Yu, C. Fang-Yen, K. Badizadegan, R. R. Dasari, and M. S. Feld, "Field-based angleresolved light-scattering study of single live cells," Opt Lett 33, 1596-1598 (2008). 25. A. J. Devaney, "Inverse-Scattering Theory within the Rytov Approximation," Opt Lett 6, 374-376 (1981).