Abstract

The use of coherent light in conventional holographic phase microscopy (HPM) poses three major drawbacks: poor spatial resolution, weak depth sectioning, and fixed pattern noise due to unwanted diffraction. Here, we report a technique which can overcome these drawbacks, but maintains the advantage of phase microscopy - high contrast live cell imaging and 3D imaging. A speckle beam of a complex spatial pattern is used for illumination to reduce fixed pattern noise and to improve optical sectioning capability. By recording of the electric field of speckle, we demonstrate high contrast 3D live cell imaging without the need for axial scanning - neither objective lens nor sample stage. This technique has great potential in studying biological samples with improved sensitivity, resolution and optical sectioning capability.

© 2009 OSA

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References

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  1. E. Cuche, F. Bevilacqua, and C. Depeursinge, “Digital holography for quantitative phase-contrast imaging,” Opt. Lett. 24(5), 291–293 (1999).
    [CrossRef]
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    [CrossRef] [PubMed]
  3. G. Popescu, T. Ikeda, R. R. Dasari, and M. S. Feld, “Diffraction phase microscopy for quantifying cell structure and dynamics,” Opt. Lett. 31(6), 775–777 (2006).
    [CrossRef] [PubMed]
  4. Y. K. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Diffraction phase and fluorescence microscopy,” Opt. Express 14(18), 8263–8268 (2006).
    [CrossRef] [PubMed]
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  6. Y.-K. 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(37), 13730–13735 (2008).
    [CrossRef] [PubMed]
  7. G. Popescu, Y.-K. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
    [CrossRef] [PubMed]
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    [CrossRef]
  9. Y. K. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Fresnel particle tracing in three dimensions using diffraction phase microscopy,” Opt. Lett. 32(7), 811–813 (2007).
    [CrossRef] [PubMed]
  10. G. Popescu, Y.-K. Park, W. Choi, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Imaging red blood cell dynamics by quantitative phase microscopy,” Blood Cells Mol. Dis. 41(1), 10–16 (2008).
    [CrossRef] [PubMed]
  11. 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(23), 9361–9373 (2005).
    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
  13. M. G. Somekh, C. W. See, and J. Goh, “Wide field amplitude and phase confocal microscope with speckle illumination,” Opt. Commun. 174(1-4), 75–80 (2000).
    [CrossRef]
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    [CrossRef] [PubMed]
  15. C. Ventalon and J. Mertz, “Quasi-confocal fluorescence sectioning with dynamic speckle illumination,” Opt. Lett. 30(24), 3350–3352 (2005).
    [CrossRef]
  16. J. Walker, “Non-scanning confocal fluorescence microscopy using speckle illumination,” Opt. Commun. 189(4-6), 221–226 (2001).
    [CrossRef]
  17. C. Fang-Yen, S. Oh, Y.-K. 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(11), 1572–1574 (2007).
    [CrossRef] [PubMed]
  18. W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007).
    [CrossRef] [PubMed]
  19. S. Inoue, and K. R. and Spring, Video Microscopy (Plenum Press, New York and London, 1997).
  20. G. Nomarski, “Microinterféromètre différentiel à ondes polarisées,” J. Phys. Radium 16, 9S–11S (1955).
  21. J. Goodman, Introduction to Fourier optics (Roberts & Co, 2005).

2008 (3)

G. Popescu, Y.-K. Park, W. Choi, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Imaging red blood cell dynamics by quantitative phase microscopy,” Blood Cells Mol. Dis. 41(1), 10–16 (2008).
[CrossRef] [PubMed]

Y.-K. 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(37), 13730–13735 (2008).
[CrossRef] [PubMed]

G. Popescu, Y.-K. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

2007 (3)

2006 (2)

2005 (4)

2004 (3)

2001 (1)

J. Walker, “Non-scanning confocal fluorescence microscopy using speckle illumination,” Opt. Commun. 189(4-6), 221–226 (2001).
[CrossRef]

2000 (1)

M. G. Somekh, C. W. See, and J. Goh, “Wide field amplitude and phase confocal microscope with speckle illumination,” Opt. Commun. 174(1-4), 75–80 (2000).
[CrossRef]

1999 (1)

1955 (1)

G. Nomarski, “Microinterféromètre différentiel à ondes polarisées,” J. Phys. Radium 16, 9S–11S (1955).

Badizadegan, K.

G. Popescu, Y.-K. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

G. Popescu, Y.-K. Park, W. Choi, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Imaging red blood cell dynamics by quantitative phase microscopy,” Blood Cells Mol. Dis. 41(1), 10–16 (2008).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007).
[CrossRef] [PubMed]

Y. K. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Fresnel particle tracing in three dimensions using diffraction phase microscopy,” Opt. Lett. 32(7), 811–813 (2007).
[CrossRef] [PubMed]

Y. K. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Diffraction phase and fluorescence microscopy,” Opt. Express 14(18), 8263–8268 (2006).
[CrossRef] [PubMed]

G. Popescu, T. Ikeda, C. A. Best, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Erythrocyte structure and dynamics quantified by Hilbert phase microscopy,” J. Biomed. Opt. 10(6), 060503 (2005).
[CrossRef]

G. Popescu, L. P. Deflores, J. C. Vaughan, K. Badizadegan, H. Iwai, R. R. Dasari, and M. S. Feld, “Fourier phase microscopy for investigation of biological structures and dynamics,” Opt. Lett. 29(21), 2503–2505 (2004).
[CrossRef] [PubMed]

Best, C. A.

G. Popescu, T. Ikeda, C. A. Best, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Erythrocyte structure and dynamics quantified by Hilbert phase microscopy,” J. Biomed. Opt. 10(6), 060503 (2005).
[CrossRef]

Best-Popescu, C.

G. Popescu, Y.-K. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

Bevilacqua, F.

Choi, W.

Y.-K. 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(37), 13730–13735 (2008).
[CrossRef] [PubMed]

G. Popescu, Y.-K. Park, W. Choi, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Imaging red blood cell dynamics by quantitative phase microscopy,” Blood Cells Mol. Dis. 41(1), 10–16 (2008).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007).
[CrossRef] [PubMed]

C. Fang-Yen, S. Oh, Y.-K. 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(11), 1572–1574 (2007).
[CrossRef] [PubMed]

Cuche, E.

Dasari, R. R.

G. Popescu, Y.-K. Park, W. Choi, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Imaging red blood cell dynamics by quantitative phase microscopy,” Blood Cells Mol. Dis. 41(1), 10–16 (2008).
[CrossRef] [PubMed]

G. Popescu, Y.-K. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007).
[CrossRef] [PubMed]

Y. K. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Fresnel particle tracing in three dimensions using diffraction phase microscopy,” Opt. Lett. 32(7), 811–813 (2007).
[CrossRef] [PubMed]

C. Fang-Yen, S. Oh, Y.-K. 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(11), 1572–1574 (2007).
[CrossRef] [PubMed]

Y. K. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Diffraction phase and fluorescence microscopy,” Opt. Express 14(18), 8263–8268 (2006).
[CrossRef] [PubMed]

G. Popescu, T. Ikeda, R. R. Dasari, and M. S. Feld, “Diffraction phase microscopy for quantifying cell structure and dynamics,” Opt. Lett. 31(6), 775–777 (2006).
[CrossRef] [PubMed]

G. Popescu, T. Ikeda, C. A. Best, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Erythrocyte structure and dynamics quantified by Hilbert phase microscopy,” J. Biomed. Opt. 10(6), 060503 (2005).
[CrossRef]

G. Popescu, L. P. Deflores, J. C. Vaughan, K. Badizadegan, H. Iwai, R. R. Dasari, and M. S. Feld, “Fourier phase microscopy for investigation of biological structures and dynamics,” Opt. Lett. 29(21), 2503–2505 (2004).
[CrossRef] [PubMed]

Deflores, L.

G. Popescu, Y.-K. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

Deflores, L. P.

Depeursinge, C.

Diez-Silva, M.

Y.-K. 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(37), 13730–13735 (2008).
[CrossRef] [PubMed]

Dubois, F.

Emery, Y.

Fang-Yen, C.

Feld, M. S.

G. Popescu, Y.-K. Park, W. Choi, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Imaging red blood cell dynamics by quantitative phase microscopy,” Blood Cells Mol. Dis. 41(1), 10–16 (2008).
[CrossRef] [PubMed]

Y.-K. 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(37), 13730–13735 (2008).
[CrossRef] [PubMed]

G. Popescu, Y.-K. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007).
[CrossRef] [PubMed]

Y. K. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Fresnel particle tracing in three dimensions using diffraction phase microscopy,” Opt. Lett. 32(7), 811–813 (2007).
[CrossRef] [PubMed]

C. Fang-Yen, S. Oh, Y.-K. 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(11), 1572–1574 (2007).
[CrossRef] [PubMed]

G. Popescu, T. Ikeda, R. R. Dasari, and M. S. Feld, “Diffraction phase microscopy for quantifying cell structure and dynamics,” Opt. Lett. 31(6), 775–777 (2006).
[CrossRef] [PubMed]

Y. K. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Diffraction phase and fluorescence microscopy,” Opt. Express 14(18), 8263–8268 (2006).
[CrossRef] [PubMed]

G. Popescu, T. Ikeda, C. A. Best, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Erythrocyte structure and dynamics quantified by Hilbert phase microscopy,” J. Biomed. Opt. 10(6), 060503 (2005).
[CrossRef]

G. Popescu, L. P. Deflores, J. C. Vaughan, K. Badizadegan, H. Iwai, R. R. Dasari, and M. S. Feld, “Fourier phase microscopy for investigation of biological structures and dynamics,” Opt. Lett. 29(21), 2503–2505 (2004).
[CrossRef] [PubMed]

Goh, J.

M. G. Somekh, C. W. See, and J. Goh, “Wide field amplitude and phase confocal microscope with speckle illumination,” Opt. Commun. 174(1-4), 75–80 (2000).
[CrossRef]

Ikeda, T.

G. Popescu, T. Ikeda, R. R. Dasari, and M. S. Feld, “Diffraction phase microscopy for quantifying cell structure and dynamics,” Opt. Lett. 31(6), 775–777 (2006).
[CrossRef] [PubMed]

G. Popescu, T. Ikeda, C. A. Best, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Erythrocyte structure and dynamics quantified by Hilbert phase microscopy,” J. Biomed. Opt. 10(6), 060503 (2005).
[CrossRef]

Istasse, E.

Iwai, H.

Kim, M.

Lo, C. M.

Lue, N.

G. Popescu, Y.-K. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007).
[CrossRef] [PubMed]

Lykotrafitis, G.

Y.-K. 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(37), 13730–13735 (2008).
[CrossRef] [PubMed]

Magistretti, P.

Mann, C.

Marquet, P.

Mertz, J.

Minetti, C.

Monnom, O.

Nomarski, G.

G. Nomarski, “Microinterféromètre différentiel à ondes polarisées,” J. Phys. Radium 16, 9S–11S (1955).

Oh, S.

Park, Y. K.

Park, Y.-K.

G. Popescu, Y.-K. Park, W. Choi, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Imaging red blood cell dynamics by quantitative phase microscopy,” Blood Cells Mol. Dis. 41(1), 10–16 (2008).
[CrossRef] [PubMed]

Y.-K. 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(37), 13730–13735 (2008).
[CrossRef] [PubMed]

G. Popescu, Y.-K. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

C. Fang-Yen, S. Oh, Y.-K. 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(11), 1572–1574 (2007).
[CrossRef] [PubMed]

Pitter, M. C.

Popescu, G.

G. Popescu, Y.-K. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

Y.-K. 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(37), 13730–13735 (2008).
[CrossRef] [PubMed]

G. Popescu, Y.-K. Park, W. Choi, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Imaging red blood cell dynamics by quantitative phase microscopy,” Blood Cells Mol. Dis. 41(1), 10–16 (2008).
[CrossRef] [PubMed]

Y. K. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Fresnel particle tracing in three dimensions using diffraction phase microscopy,” Opt. Lett. 32(7), 811–813 (2007).
[CrossRef] [PubMed]

Y. K. Park, G. Popescu, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Diffraction phase and fluorescence microscopy,” Opt. Express 14(18), 8263–8268 (2006).
[CrossRef] [PubMed]

G. Popescu, T. Ikeda, R. R. Dasari, and M. S. Feld, “Diffraction phase microscopy for quantifying cell structure and dynamics,” Opt. Lett. 31(6), 775–777 (2006).
[CrossRef] [PubMed]

G. Popescu, T. Ikeda, C. A. Best, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Erythrocyte structure and dynamics quantified by Hilbert phase microscopy,” J. Biomed. Opt. 10(6), 060503 (2005).
[CrossRef]

G. Popescu, L. P. Deflores, J. C. Vaughan, K. Badizadegan, H. Iwai, R. R. Dasari, and M. S. Feld, “Fourier phase microscopy for investigation of biological structures and dynamics,” Opt. Lett. 29(21), 2503–2505 (2004).
[CrossRef] [PubMed]

Rappaz, B.

Requena, M. L.

See, C. W.

M. C. Pitter, C. W. See, and M. G. Somekh, “Full-field heterodyne interference microscope with spatially incoherent illumination,” Opt. Lett. 29(11), 1200–1202 (2004).
[CrossRef] [PubMed]

M. G. Somekh, C. W. See, and J. Goh, “Wide field amplitude and phase confocal microscope with speckle illumination,” Opt. Commun. 174(1-4), 75–80 (2000).
[CrossRef]

Seung, H. S.

Somekh, M. G.

M. C. Pitter, C. W. See, and M. G. Somekh, “Full-field heterodyne interference microscope with spatially incoherent illumination,” Opt. Lett. 29(11), 1200–1202 (2004).
[CrossRef] [PubMed]

M. G. Somekh, C. W. See, and J. Goh, “Wide field amplitude and phase confocal microscope with speckle illumination,” Opt. Commun. 174(1-4), 75–80 (2000).
[CrossRef]

Song, S.

Suresh, S.

Y.-K. 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(37), 13730–13735 (2008).
[CrossRef] [PubMed]

Vaughan, J. C.

Ventalon, C.

Walker, J.

J. Walker, “Non-scanning confocal fluorescence microscopy using speckle illumination,” Opt. Commun. 189(4-6), 221–226 (2001).
[CrossRef]

Yu, L.

Am. J. Physiol. Cell Physiol. (1)

G. Popescu, Y.-K. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

Appl. Opt. (1)

Blood Cells Mol. Dis. (1)

G. Popescu, Y.-K. Park, W. Choi, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Imaging red blood cell dynamics by quantitative phase microscopy,” Blood Cells Mol. Dis. 41(1), 10–16 (2008).
[CrossRef] [PubMed]

J. Biomed. Opt. (1)

G. Popescu, T. Ikeda, C. A. Best, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Erythrocyte structure and dynamics quantified by Hilbert phase microscopy,” J. Biomed. Opt. 10(6), 060503 (2005).
[CrossRef]

J. Phys. Radium (1)

G. Nomarski, “Microinterféromètre différentiel à ondes polarisées,” J. Phys. Radium 16, 9S–11S (1955).

Nat. Methods (1)

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007).
[CrossRef] [PubMed]

Opt. Commun. (2)

J. Walker, “Non-scanning confocal fluorescence microscopy using speckle illumination,” Opt. Commun. 189(4-6), 221–226 (2001).
[CrossRef]

M. G. Somekh, C. W. See, and J. Goh, “Wide field amplitude and phase confocal microscope with speckle illumination,” Opt. Commun. 174(1-4), 75–80 (2000).
[CrossRef]

Opt. Express (3)

Opt. Lett. (7)

Proc. Natl. Acad. Sci. U.S.A. (1)

Y.-K. 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(37), 13730–13735 (2008).
[CrossRef] [PubMed]

Other (2)

S. Inoue, and K. R. and Spring, Video Microscopy (Plenum Press, New York and London, 1997).

J. Goodman, Introduction to Fourier optics (Roberts & Co, 2005).

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Figures (5)

Fig. 1
Fig. 1

(a) Principle of speckle field holographic microscopy. A holographic diffuser (H) is illuminated with a laser beam. The complex E-field of the scattered beam is measured from the interference image (hologram 1) after interference with a reference beam. After introducing a sample (S), another interference image is measured (hologram 2). (b-c) amplitude (b) and phase (c) of the E-field without the sample, (d-e) amplitude (d) and phase (e) with the sample. (f-g) Amplitude (f) and phase (g) are images of the sample, a 10 μm polystyrene bead. Scale bar, 5 μm. Colorbars indicate arbitrary units for the amplitude and radians for the phase, respectively.

Fig. 2
Fig. 2

(a) Experimental setup. BS: beam splitter, GM: galvanometer mirrors, H: a holographic diffuser, S: sample, Condenser: Nikon 1.4NA, Camera: (Fastcam 1024 PCI, Photron), AOM1,2: acousto-optic modulators for temporal modulation of interference by shifting the frequency of the reference beam. (b-e) Averaged phase images where N is the number of speckle fields used for averaging. Scale bar indicates 5 μm and color bar indicates radian. (f) Standard deviation of phase noise as a function of the number of speckle fields used for averaging. Thin lines indicate the averaged phase for an area of 4 × 4 μm2 located outside of the sample. Thick line shows the averaged phase for 6 different areas.

Fig. 3
Fig. 3

(a) Amplitude and (b) phase images of the E-field measured for a 10 μm polystyrene bead with a conventional HPM. (d) Amplitude and (e) phase images of the same sample measure by SDHM. (c, f), Amplitude images of USAF resolution (group 9) target measured by conventional HPM and SDHM, respectively. Scale bar indicates 5 μm, and colorbars indicate arbitrary unit for amplitude images and radians for the phase images, respectively. For SDHM, 100 raw images are processed to reduce singularity points.

Fig. 4
Fig. 4

Speckle illumination increases lateral and axial resolving power. (a-d) Phase maps of polystyrene beads (200nm diameter) measured with a conventional HPM. (e-h) Phase maps measured with a speckle illumination of the same samples in (a-d). Scale bar indicates 500 nm. (i) Lateral point spread functions of a conventional HPM (blue line) and a speckle illumination (red line). (j) Axial point spread functions of a conventional HPM (blue line) and a speckle illumination (red line). For SDHM, 50 raw E-field images are processed to reduce singularity points.

Fig. 5
Fig. 5

3D live cell imaging using SDHM. (a-b) Amplitude maps of a HeLa cell measured with (a) HPM and (b) SDHM, respectively. Color bar indicates arbitrary unit of amplitude. (c-d) Quantitative phase maps of the same cell imaged using HPM and SDHM, respectively. Color bar indicates phase in radian. (e) Emulated differential interference contract (DIC) images of the same cell taken by HPM. (f-h) Emulated DIC images by SDHM; (f) and (h) show numerically propagated DIC images 0.5 μm below and above form the original focus plane (g), respectively. Arrows indicate sub-cellular particles or vesicles focused on the specific focal planes. Scale bar indicates 5 μm. For SDHM, 50 raw E-field images are processed to reduce singularity points.

Equations (1)

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d=1.22λ/(NAO+NAI),

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