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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 20, Iss. 6 — Mar. 12, 2012
  • pp: 6737–6745

Gradient field microscopy of unstained specimens

Taewoo Kim, Shamira Sridharan, and Gabriel Popescu  »View Author Affiliations

Optics Express, Vol. 20, Issue 6, pp. 6737-6745 (2012)

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We present a phase derivative microscopy technique referred to as gradient field microscopy (GFM), which provides the first-order derivatives of the phase associated with an optical field passing through a transparent specimen. GFM utilizes spatial light modulation at the Fourier plane of a bright field microscope to optically obtain the derivatives of the phase and increase the contrast of the final image. The controllable spatial modulation pattern allows us to obtain both one component of the field gradient (derivative along one direction) and the gradient intensity, which offers some advantages over the regular differential interference contrast (DIC) microscopy. Most importantly, unlike DIC, GFM does not use polarizing optics and, thus, it is applicable to birefringent samples. We demonstrate these features of GFM with studies of static and dynamic biological cells (HeLa cells and red blood cells). We show that GFM is capable of qualitatively providing information about cell membrane fluctuations. Specifically, we captured the disappearance of the bending mode of fluctuations in osmotically swollen red blood cells.

© 2012 OSA

OCIS Codes
(170.0180) Medical optics and biotechnology : Microscopy
(170.1530) Medical optics and biotechnology : Cell analysis
(170.1650) Medical optics and biotechnology : Coherence imaging

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: February 2, 2012
Revised Manuscript: March 2, 2012
Manuscript Accepted: March 3, 2012
Published: March 7, 2012

Virtual Issues
Vol. 7, Iss. 5 Virtual Journal for Biomedical Optics

Taewoo Kim, Shamira Sridharan, and Gabriel Popescu, "Gradient field microscopy of unstained specimens," Opt. Express 20, 6737-6745 (2012)

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  1. G. Popescu, Quantitative phase imaging of cells and tissues (McGraw-Hill, 2011).
  2. “Milestones in light microscopy,” Nat. Cell Biol.11(10), 1165–1165 (2009). [CrossRef] [PubMed]
  3. F. Zernike, “How I discovered phase contrast,” Science121(3141), 345–349 (1955). [CrossRef] [PubMed]
  4. M. G. Nomarski, “Microinterferometre differentiel a ondes polarisees,” J. Phys. (Paris)16, S9–S13 (1955).
  5. D. K. Hamilton and C. J. R. Sheppard, “Differential phase-contrast in scanning optical microscopy,” J Microsc. (Oxford)133(1), 27–39 (1984). [CrossRef]
  6. T. Wilson, “Enhanced differential phase-contrast imaging in scanning microscopy using a quadrant detector,” Optik (Stuttg.)80, 167–170 (1988).
  7. A. Ahn, C. H. Yang, A. Wax, G. Popescu, C. Fang-Yen, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Harmonic phase-dispersion microscope with a Mach-Zehnder interferometer,” Appl. Opt.44(7), 1188–1190 (2005). [CrossRef] [PubMed]
  8. C. H. Yang, A. Wax, I. Georgakoudi, E. B. Hanlon, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Interferometric phase-dispersion microscopy,” Opt. Lett.25(20), 1526–1528 (2000). [CrossRef] [PubMed]
  9. C. K. Hitzenberger and A. F. Fercher, “Differential phase contrast in optical coherence tomography,” Opt. Lett.24(9), 622–624 (1999). [CrossRef] [PubMed]
  10. M. Lew, X. Q. Cui, X. Heng, and C. H. Yang, “Interference of a four-hole aperture for on-chip quantitative two-dimensional differential phase imaging,” Opt. Lett.32(20), 2963–2965 (2007). [CrossRef] [PubMed]
  11. C. Rylander, T. E. Milner, K. R. Diller, and A. J. Welch, “Spatial refractive index determination of single cells using differential phase optical low-coherence reflectometry,” Lasers Surg. Med., 5–5 (2003).
  12. T. J. McIntyre, C. Maurer, S. Bernet, and M. Ritsch-Marte, “Differential interference contrast imaging using a spatial light modulator,” Opt. Lett.34(19), 2988–2990 (2009). [CrossRef] [PubMed]
  13. S. H. Lee and D. P. Casasent, Optical Information Processing: Fundamentals (Springer-Verlag, 1981).
  14. T. Kim and G. Popescu, “Laplace field microscopy for label-free imaging of dynamic biological structures,” Opt. Lett.36(23), 4704–4706 (2011). [CrossRef] [PubMed]
  15. W. C. Stewart, “Differential phase-contrast with an extended illumination source,” J. Opt. Soc. Am.66(8), 813–818 (1976). [CrossRef]
  16. B. Kachar, “Asymmetric illumination contrast: a method of image formation for video light microscopy,” Science227(4688), 766–768 (1985). [CrossRef] [PubMed]
  17. S. B. Mehta and C. J. R. Sheppard, “Quantitative phase-gradient imaging at high resolution with asymmetric illumination-based differential phase contrast,” Opt. Lett.34(13), 1924–1926 (2009). [CrossRef] [PubMed]
  18. R. Yi, K. K. Chu, and J. Mertz, “Graded-field microscopy with white light,” Opt. Express14(12), 5191–5200 (2006). [CrossRef] [PubMed]
  19. G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Observation of dynamic subdomains in red blood cells,” J. Biomed. Opt.11(4), 040503 (2006). [CrossRef] [PubMed]
  20. 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(21), 218101 (2006). [CrossRef] [PubMed]
  21. H. F. Ding, Z. Wang, F. Nguyen, S. A. Boppart, and G. Popescu, “Fourier transform light scattering of inhomogeneous and dynamic structures,” Phys. Rev. Lett.101(23), 238102 (2008). [CrossRef] [PubMed]
  22. Y. K. Park, C. A. Best, T. Auth, N. S. Gov, S. A. Safran, G. Popescu, S. Suresh, and M. S. Feld, “Metabolic remodeling of the human red blood cell membrane,” Proc. Natl. Acad. Sci. U.S.A.107(4), 1289–1294 (2010). [CrossRef] [PubMed]
  23. Y. K. Park, C. A. Best, K. Badizadegan, R. R. Dasari, M. S. Feld, T. Kuriabova, M. L. Henle, A. J. Levine, and G. Popescu, “Measurement of red blood cell mechanics during morphological changes,” Proc. Natl. Acad. Sci. U.S.A.107(15), 6731–6736 (2010). [CrossRef] [PubMed]
  24. Y. Park, C. A. Best, T. Kuriabova, M. L. Henle, M. S. Feld, A. J. Levine, and G. Popescu, “Measurement of the nonlinear elasticity of red blood cell membranes,” Phys. Rev. E Stat. Nonlin. Soft Matter Phys.83(5), 051925 (2011). [CrossRef] [PubMed]
  25. Y. K. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. S. 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(37), 13730–13735 (2008). [CrossRef] [PubMed]
  26. N. Gov, A. G. Zilman, and S. Safran, “Cytoskeleton confinement and tension of red blood cell membranes,” Phys. Rev. Lett.90(22), 228101 (2003). [CrossRef] [PubMed]

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