Abstract

We report a quantitative phase microscope based on spectral domain optical coherence tomography and line-field illumination. The line illumination allows self phase-referencing method to reject common-mode phase noise. The quantitative phase microscope also features a separate reference arm, permitting the use of high numerical aperture (NA > 1) microscope objectives for high resolution phase measurement at multiple points along the line of illumination. We demonstrate that the path-length sensitivity of the instrument can be as good as 41pm/Hz, which makes it suitable for nanometer scale study of cell motility. We present the detection of natural motions of cell surface and two-dimensional surface profiling of a HeLa cell.

© 2009 OSA

Full Article  |  PDF Article

References

  • View by:
  • |
  • |
  • |

  1. P. C. Zhang, A. M. Keleshian, and F. Sachs, “Voltage-induced membrane movement,” Nature 413(6854), 428–432 (2001).
    [CrossRef] [PubMed]
  2. E. Cuche, F. Bevilacqua, and C. Depeursinge, “Digital holography for quantitative phase-contrast imaging,” Opt. Lett. 24(5), 291–293 (1999).
    [CrossRef]
  3. 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]
  4. T. Ikeda, G. Popescu, R. R. Dasari, and M. S. Feld, “Hilbert phase microscopy for investigating fast dynamics in transparent systems,” Opt. Lett. 30(10), 1165–1167 (2005).
    [CrossRef] [PubMed]
  5. 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]
  6. J. Wu, Z. Yaqoob, X. Heng, L. M. Lee, X. Q. Cui, and C. H. Yang, “Full field phase imaging using a harmonically matched diffraction grating pair based homodyne quadrature interferometer,” Appl. Phys. Lett. 90(15), 151123 (2007).
    [CrossRef]
  7. N. Lue, W. Choi, G. Popescu, T. Ikeda, R. R. Dasari, K. Badizadegan, and M. S. Feld, “Quantitative phase imaging of live cells using fast Fourier phase microscopy,” Appl. Opt. 46(10), 1836–1842 (2007).
    [CrossRef] [PubMed]
  8. M. V. Sarunic, S. Weinberg, and J. A. Izatt, “Full-field swept-source phase microscopy,” Opt. Lett. 31(10), 1462–1464 (2006).
    [CrossRef] [PubMed]
  9. M. A. Choma, A. K. Ellerbee, C. H. Yang, T. L. Creazzo, and J. A. Izatt, “Spectral-domain phase microscopy,” Opt. Lett. 30(10), 1162–1164 (2005).
    [CrossRef] [PubMed]
  10. C. Joo, T. Akkin, B. Cense, B. H. Park, and J. F. de Boer, “Spectral-domain optical coherence phase microscopy for quantitative phase-contrast imaging,” Opt. Lett. 30(16), 2131–2133 (2005).
    [CrossRef] [PubMed]
  11. T. Yamauchi, H. Iwai, M. Miwa, and Y. Yamashita, “Low-coherent quantitative phase microscope for nanometer-scale measurement of living cells morphology,” Opt. Express 16(16), 12227–12238 (2008).
    [CrossRef] [PubMed]
  12. F. Lexer, C. K. Hitzenberger, A. F. Fercher, and M. Kulhavy, “Wavelength-tuning interferometry of intraocular distances,” Appl. Opt. 36(25), 6548–6553 (1997).
    [CrossRef]
  13. G. Hausler and M. W. Lindner, “"Coherence Radar” and “Spectral Radar” - New Tools for Dermatological Diagnosis,” J. Biomed. Opt. 3(1), 21–31 (1998).
    [CrossRef]
  14. A. K. Ellerbee and J. A. Izatt, “Phase retrieval in low-coherence interferometric microscopy,” Opt. Lett. 32(4), 388–390 (2007).
    [CrossRef] [PubMed]
  15. T. Endo, Y. Yasuno, S. Makita, M. Itoh, and T. Yatagai, “Profilometry with line-field Fourier-domain interferometry,” Opt. Express 13(3), 695–701 (2005).
    [CrossRef] [PubMed]
  16. B. Grajciar, M. Pircher, A. F. Fercher, and R. A. Leitgeb, “Parallel Fourier domain optical coherence tomography for in vivo measurement of the human eye,” Opt. Express 13(4), 1131–1137 (2005).
    [CrossRef] [PubMed]
  17. Y. Yasuno, T. Endo, S. Makita, G. Aoki, M. Itoh, and T. Yatagai, “Three-dimensional line-field Fourier domain optical coherence tomography for in vivo dermatological investigation,” J. Biomed. Opt. 11(1), 014014 (2006).
    [CrossRef] [PubMed]
  18. Y. Nakamura, S. Makita, M. Yamanari, M. Itoh, T. Yatagai, and Y. Yasuno, “High-speed three-dimensional human retinal imaging by line-field spectral domain optical coherence tomography,” Opt. Express 15(12), 7103–7116 (2007).
    [CrossRef] [PubMed]
  19. G. Popescu, Y. Park, R. R. Dasari, K. Badizadegan, and M. S. Feld, “Coherence properties of red blood cell membrane motions,” Phys. Rev. E Stat. Nonlin. Soft Matter Phys. 76(3 Pt 1), 031902 (2007).
    [CrossRef] [PubMed]
  20. 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]
  21. T. J. Flynn, “Two-dimensional phase unwrapping with minimum weighted discontinuity,” J. Opt. Soc. Am. A 14(10), 2692–2701 (1997).
    [CrossRef]

2008 (1)

2007 (6)

J. Wu, Z. Yaqoob, X. Heng, L. M. Lee, X. Q. Cui, and C. H. Yang, “Full field phase imaging using a harmonically matched diffraction grating pair based homodyne quadrature interferometer,” Appl. Phys. Lett. 90(15), 151123 (2007).
[CrossRef]

G. Popescu, Y. Park, R. R. Dasari, K. Badizadegan, and M. S. Feld, “Coherence properties of red blood cell membrane motions,” Phys. Rev. E Stat. Nonlin. Soft Matter Phys. 76(3 Pt 1), 031902 (2007).
[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]

A. K. Ellerbee and J. A. Izatt, “Phase retrieval in low-coherence interferometric microscopy,” Opt. Lett. 32(4), 388–390 (2007).
[CrossRef] [PubMed]

N. Lue, W. Choi, G. Popescu, T. Ikeda, R. R. Dasari, K. Badizadegan, and M. S. Feld, “Quantitative phase imaging of live cells using fast Fourier phase microscopy,” Appl. Opt. 46(10), 1836–1842 (2007).
[CrossRef] [PubMed]

Y. Nakamura, S. Makita, M. Yamanari, M. Itoh, T. Yatagai, and Y. Yasuno, “High-speed three-dimensional human retinal imaging by line-field spectral domain optical coherence tomography,” Opt. Express 15(12), 7103–7116 (2007).
[CrossRef] [PubMed]

2006 (3)

2005 (5)

2004 (1)

2001 (1)

P. C. Zhang, A. M. Keleshian, and F. Sachs, “Voltage-induced membrane movement,” Nature 413(6854), 428–432 (2001).
[CrossRef] [PubMed]

1999 (1)

1998 (1)

G. Hausler and M. W. Lindner, “"Coherence Radar” and “Spectral Radar” - New Tools for Dermatological Diagnosis,” J. Biomed. Opt. 3(1), 21–31 (1998).
[CrossRef]

1997 (2)

F. Lexer, C. K. Hitzenberger, A. F. Fercher, and M. Kulhavy, “Wavelength-tuning interferometry of intraocular distances,” Appl. Opt. 36(25), 6548–6553 (1997).
[CrossRef]

T. J. Flynn, “Two-dimensional phase unwrapping with minimum weighted discontinuity,” J. Opt. Soc. Am. A 14(10), 2692–2701 (1997).
[CrossRef]

Akkin, T.

Aoki, G.

Y. Yasuno, T. Endo, S. Makita, G. Aoki, M. Itoh, and T. Yatagai, “Three-dimensional line-field Fourier domain optical coherence tomography for in vivo dermatological investigation,” J. Biomed. Opt. 11(1), 014014 (2006).
[CrossRef] [PubMed]

Badizadegan, K.

G. Popescu, Y. Park, R. R. Dasari, K. Badizadegan, and M. S. Feld, “Coherence properties of red blood cell membrane motions,” Phys. Rev. E Stat. Nonlin. Soft Matter Phys. 76(3 Pt 1), 031902 (2007).
[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]

N. Lue, W. Choi, G. Popescu, T. Ikeda, R. R. Dasari, K. Badizadegan, and M. S. Feld, “Quantitative phase imaging of live cells using fast Fourier phase microscopy,” Appl. Opt. 46(10), 1836–1842 (2007).
[CrossRef] [PubMed]

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]

Bevilacqua, F.

Cense, B.

Choi, W.

Choma, M. A.

Creazzo, T. L.

Cuche, E.

Cui, X. Q.

J. Wu, Z. Yaqoob, X. Heng, L. M. Lee, X. Q. Cui, and C. H. Yang, “Full field phase imaging using a harmonically matched diffraction grating pair based homodyne quadrature interferometer,” Appl. Phys. Lett. 90(15), 151123 (2007).
[CrossRef]

Dasari, R. R.

de Boer, J. F.

Deflores, L. P.

Depeursinge, C.

Ellerbee, A. K.

Endo, T.

Y. Yasuno, T. Endo, S. Makita, G. Aoki, M. Itoh, and T. Yatagai, “Three-dimensional line-field Fourier domain optical coherence tomography for in vivo dermatological investigation,” J. Biomed. Opt. 11(1), 014014 (2006).
[CrossRef] [PubMed]

T. Endo, Y. Yasuno, S. Makita, M. Itoh, and T. Yatagai, “Profilometry with line-field Fourier-domain interferometry,” Opt. Express 13(3), 695–701 (2005).
[CrossRef] [PubMed]

Fang-Yen, C.

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]

Feld, M. S.

Fercher, A. F.

Flynn, T. J.

T. J. Flynn, “Two-dimensional phase unwrapping with minimum weighted discontinuity,” J. Opt. Soc. Am. A 14(10), 2692–2701 (1997).
[CrossRef]

Grajciar, B.

Hausler, G.

G. Hausler and M. W. Lindner, “"Coherence Radar” and “Spectral Radar” - New Tools for Dermatological Diagnosis,” J. Biomed. Opt. 3(1), 21–31 (1998).
[CrossRef]

Heng, X.

J. Wu, Z. Yaqoob, X. Heng, L. M. Lee, X. Q. Cui, and C. H. Yang, “Full field phase imaging using a harmonically matched diffraction grating pair based homodyne quadrature interferometer,” Appl. Phys. Lett. 90(15), 151123 (2007).
[CrossRef]

Hitzenberger, C. K.

Ikeda, T.

Itoh, M.

Iwai, H.

Izatt, J. A.

Joo, C.

Keleshian, A. M.

P. C. Zhang, A. M. Keleshian, and F. Sachs, “Voltage-induced membrane movement,” Nature 413(6854), 428–432 (2001).
[CrossRef] [PubMed]

Kulhavy, M.

Lee, L. M.

J. Wu, Z. Yaqoob, X. Heng, L. M. Lee, X. Q. Cui, and C. H. Yang, “Full field phase imaging using a harmonically matched diffraction grating pair based homodyne quadrature interferometer,” Appl. Phys. Lett. 90(15), 151123 (2007).
[CrossRef]

Leitgeb, R. A.

Lexer, F.

Lindner, M. W.

G. Hausler and M. W. Lindner, “"Coherence Radar” and “Spectral Radar” - New Tools for Dermatological Diagnosis,” J. Biomed. Opt. 3(1), 21–31 (1998).
[CrossRef]

Lue, N.

Makita, S.

Miwa, M.

Nakamura, Y.

Oh, S.

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]

Park, B. H.

Park, Y.

G. Popescu, Y. Park, R. R. Dasari, K. Badizadegan, and M. S. Feld, “Coherence properties of red blood cell membrane motions,” Phys. Rev. E Stat. Nonlin. Soft Matter Phys. 76(3 Pt 1), 031902 (2007).
[CrossRef] [PubMed]

Pircher, M.

Popescu, G.

Sachs, F.

P. C. Zhang, A. M. Keleshian, and F. Sachs, “Voltage-induced membrane movement,” Nature 413(6854), 428–432 (2001).
[CrossRef] [PubMed]

Sarunic, M. V.

Vaughan, J. C.

Weinberg, S.

Wu, J.

J. Wu, Z. Yaqoob, X. Heng, L. M. Lee, X. Q. Cui, and C. H. Yang, “Full field phase imaging using a harmonically matched diffraction grating pair based homodyne quadrature interferometer,” Appl. Phys. Lett. 90(15), 151123 (2007).
[CrossRef]

Yamanari, M.

Yamashita, Y.

Yamauchi, T.

Yang, C. H.

J. Wu, Z. Yaqoob, X. Heng, L. M. Lee, X. Q. Cui, and C. H. Yang, “Full field phase imaging using a harmonically matched diffraction grating pair based homodyne quadrature interferometer,” Appl. Phys. Lett. 90(15), 151123 (2007).
[CrossRef]

M. A. Choma, A. K. Ellerbee, C. H. Yang, T. L. Creazzo, and J. A. Izatt, “Spectral-domain phase microscopy,” Opt. Lett. 30(10), 1162–1164 (2005).
[CrossRef] [PubMed]

Yaqoob, Z.

J. Wu, Z. Yaqoob, X. Heng, L. M. Lee, X. Q. Cui, and C. H. Yang, “Full field phase imaging using a harmonically matched diffraction grating pair based homodyne quadrature interferometer,” Appl. Phys. Lett. 90(15), 151123 (2007).
[CrossRef]

Yasuno, Y.

Yatagai, T.

Zhang, P. C.

P. C. Zhang, A. M. Keleshian, and F. Sachs, “Voltage-induced membrane movement,” Nature 413(6854), 428–432 (2001).
[CrossRef] [PubMed]

,” J. Opt. Soc. Am. A (1)

T. J. Flynn, “Two-dimensional phase unwrapping with minimum weighted discontinuity,” J. Opt. Soc. Am. A 14(10), 2692–2701 (1997).
[CrossRef]

Appl. Opt. (2)

Appl. Phys. Lett. (1)

J. Wu, Z. Yaqoob, X. Heng, L. M. Lee, X. Q. Cui, and C. H. Yang, “Full field phase imaging using a harmonically matched diffraction grating pair based homodyne quadrature interferometer,” Appl. Phys. Lett. 90(15), 151123 (2007).
[CrossRef]

J. Biomed. Opt. (2)

G. Hausler and M. W. Lindner, “"Coherence Radar” and “Spectral Radar” - New Tools for Dermatological Diagnosis,” J. Biomed. Opt. 3(1), 21–31 (1998).
[CrossRef]

Y. Yasuno, T. Endo, S. Makita, G. Aoki, M. Itoh, and T. Yatagai, “Three-dimensional line-field Fourier domain optical coherence tomography for in vivo dermatological investigation,” J. Biomed. Opt. 11(1), 014014 (2006).
[CrossRef] [PubMed]

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]

Nature (1)

P. C. Zhang, A. M. Keleshian, and F. Sachs, “Voltage-induced membrane movement,” Nature 413(6854), 428–432 (2001).
[CrossRef] [PubMed]

Opt. Express (4)

Opt. Lett. (8)

Phys. Rev. E Stat. Nonlin. Soft Matter Phys. (1)

G. Popescu, Y. Park, R. R. Dasari, K. Badizadegan, and M. S. Feld, “Coherence properties of red blood cell membrane motions,” Phys. Rev. E Stat. Nonlin. Soft Matter Phys. 76(3 Pt 1), 031902 (2007).
[CrossRef] [PubMed]

Cited By

OSA participates in CrossRef's Cited-By Linking service. Citing articles from OSA journals and other participating publishers are listed here.

Alert me when this article is cited.


Figures (4)

Fig. 1
Fig. 1

(a) Schematic of line-field phase microscope (LFPM). (b,c) Horizontal and vertical perspectives, respectively, of the LFPM. (d) Typical 2-D recorded interferogram illustrating spectral and spatial measurements along the two orthogonal directions of the 2-D spectrometer. SMF: single mode fiber, Li: ith spherical lens, CL: cylindrical lens, BS: beam splitter, S: slit, G: diffraction grating.

Fig. 2
Fig. 2

Phase measurements from a fixed location on the surface of a 10 microns diameter latex microsphere (a) before and (b) after common-mode noise removal. (c) Time traces of the measured phase at one point on the line-illumination with and without self-phase referencing.

Fig. 3
Fig. 3

(a) Setup to the experimentally measure the phase sensitivity of the LFPM. (b) shows the log-log plot of the peak displacement of piezo actuator measured versus peak sinusoidal drive voltage using LFPM as well as standard interferometry. The measurement sensitivity, i.e., the minimum motion detected by the LFPM, was determined to be 41 pm/Hz .

Fig. 4
Fig. 4

(a) shows spatially averaged cell membrane fluctuations, Δφ , of a HeLa cell before and after chemical fixation. Note the reduced cellular motions after cross-linking of cellular proteins by formalin fixation. Post-formalin residual motions likely represent residual thermal motion of the cell surface. (b) 2-D surface profile of a HeLa cell measured by displacing the cell across the line-illumination. A total phase of more than 100 radians was measured with respect to the glass coverslip. Assuming the average index of the cell as 1.37, the overall cell height was estimated as 5 μm.

Equations (1)

Equations on this page are rendered with MathJax. Learn more.

Iint(l,m)=IR+IS(m)+2IRIS(m)cos[2nk(l)(zRzS(m))]

Metrics