Abstract

We present a high-speed synthetic aperture microscopy for quantitative phase imaging of live biological cells. We measure 361 complex amplitude images of an object with various directions of illumination covering an NA of 0.8 in less than one-thirteenth of a second and then combine the images with a phase-referencing method to create a synthesized phase image. Because of the increased depth selectivity, artifacts from diffraction that are typically present in coherent imaging are significantly suppressed, and lateral resolution of phase imaging is improved. We use the instrument to demonstrate high-quality phase imaging of live cells, both static and dynamic, and thickness measurements of a nanoscale cholesterol helical ribbon.

© 2011 Optical Society of America

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2010 (1)

2009 (2)

R. Fiolka, K. Wicker, R. Heintzmann, and A. Stemmer, Opt. Express 17, 12407 (2009).
[CrossRef] [PubMed]

B. Khaykovich, N. Kozlova, W. Choi, A. Lomakin, C. Hossain, Y. Sung, R. R. Dasari, M. S. Feld, and G. B. Benedek, Proc. Natl. Acad. Sci. USA 106, 15663 (2009).
[CrossRef] [PubMed]

2008 (1)

M. Debailleul, B. Simon, V. Georges, O. Haeberle, and V. Lauer, Meas. Sci. Technol. 19, 074009 (2008).
[CrossRef]

2007 (3)

2005 (1)

2003 (1)

Alexandrov, S. A.

Badizadegan, K.

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, Nat. Methods 4, 717 (2007).
[CrossRef] [PubMed]

Benedek, G. B.

B. Khaykovich, N. Kozlova, W. Choi, A. Lomakin, C. Hossain, Y. Sung, R. R. Dasari, M. S. Feld, and G. B. Benedek, Proc. Natl. Acad. Sci. USA 106, 15663 (2009).
[CrossRef] [PubMed]

Brooker, G.

Brueck, S. R. J.

Choi, W.

B. Khaykovich, N. Kozlova, W. Choi, A. Lomakin, C. Hossain, Y. Sung, R. R. Dasari, M. S. Feld, and G. B. Benedek, Proc. Natl. Acad. Sci. USA 106, 15663 (2009).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, Nat. Methods 4, 717 (2007).
[CrossRef] [PubMed]

C. Fang-Yen, S. Oh, Y. Park, W. Choi, S. Song, H. S. Seung, R. R. Dasari, and M. S. Feld, Opt. Lett. 32, 1572 (2007).
[CrossRef] [PubMed]

Cuche, E.

Dasari, R. R.

B. Khaykovich, N. Kozlova, W. Choi, A. Lomakin, C. Hossain, Y. Sung, R. R. Dasari, M. S. Feld, and G. B. Benedek, Proc. Natl. Acad. Sci. USA 106, 15663 (2009).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, Nat. Methods 4, 717 (2007).
[CrossRef] [PubMed]

C. Fang-Yen, S. Oh, Y. Park, W. Choi, S. Song, H. S. Seung, R. R. Dasari, and M. S. Feld, Opt. Lett. 32, 1572 (2007).
[CrossRef] [PubMed]

Debailleul, M.

M. Debailleul, B. Simon, V. Georges, O. Haeberle, and V. Lauer, Meas. Sci. Technol. 19, 074009 (2008).
[CrossRef]

Depeursinge, C.

Emery, Y.

Fang-Yen, C.

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, Nat. Methods 4, 717 (2007).
[CrossRef] [PubMed]

C. Fang-Yen, S. Oh, Y. Park, W. Choi, S. Song, H. S. Seung, R. R. Dasari, and M. S. Feld, Opt. Lett. 32, 1572 (2007).
[CrossRef] [PubMed]

Feld, M. S.

B. Khaykovich, N. Kozlova, W. Choi, A. Lomakin, C. Hossain, Y. Sung, R. R. Dasari, M. S. Feld, and G. B. Benedek, Proc. Natl. Acad. Sci. USA 106, 15663 (2009).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, Nat. Methods 4, 717 (2007).
[CrossRef] [PubMed]

C. Fang-Yen, S. Oh, Y. Park, W. Choi, S. Song, H. S. Seung, R. R. Dasari, and M. S. Feld, Opt. Lett. 32, 1572 (2007).
[CrossRef] [PubMed]

Fiolka, R.

Georges, V.

M. Debailleul, B. Simon, V. Georges, O. Haeberle, and V. Lauer, Meas. Sci. Technol. 19, 074009 (2008).
[CrossRef]

Goodman, J. W.

J. W. Goodman, Introduction to Fourier Optics (Roberts, 2005).

Gutzler, T.

Haeberle, O.

M. Debailleul, B. Simon, V. Georges, O. Haeberle, and V. Lauer, Meas. Sci. Technol. 19, 074009 (2008).
[CrossRef]

Heintzmann, R.

Hillman, T. R.

Hossain, C.

B. Khaykovich, N. Kozlova, W. Choi, A. Lomakin, C. Hossain, Y. Sung, R. R. Dasari, M. S. Feld, and G. B. Benedek, Proc. Natl. Acad. Sci. USA 106, 15663 (2009).
[CrossRef] [PubMed]

Indebetouw, G.

Khaykovich, B.

B. Khaykovich, N. Kozlova, W. Choi, A. Lomakin, C. Hossain, Y. Sung, R. R. Dasari, M. S. Feld, and G. B. Benedek, Proc. Natl. Acad. Sci. USA 106, 15663 (2009).
[CrossRef] [PubMed]

Kozlova, N.

B. Khaykovich, N. Kozlova, W. Choi, A. Lomakin, C. Hossain, Y. Sung, R. R. Dasari, M. S. Feld, and G. B. Benedek, Proc. Natl. Acad. Sci. USA 106, 15663 (2009).
[CrossRef] [PubMed]

Kuznetsova, Y.

Lauer, V.

M. Debailleul, B. Simon, V. Georges, O. Haeberle, and V. Lauer, Meas. Sci. Technol. 19, 074009 (2008).
[CrossRef]

Lomakin, A.

B. Khaykovich, N. Kozlova, W. Choi, A. Lomakin, C. Hossain, Y. Sung, R. R. Dasari, M. S. Feld, and G. B. Benedek, Proc. Natl. Acad. Sci. USA 106, 15663 (2009).
[CrossRef] [PubMed]

Lue, N.

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, Nat. Methods 4, 717 (2007).
[CrossRef] [PubMed]

Magistretti, P.

Marquet, P.

Oh, S.

C. Fang-Yen, S. Oh, Y. Park, W. Choi, S. Song, H. S. Seung, R. R. Dasari, and M. S. Feld, Opt. Lett. 32, 1572 (2007).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, Nat. Methods 4, 717 (2007).
[CrossRef] [PubMed]

Park, Y.

Rappaz, B.

Rosen, J.

Sampson, D. D.

Schwarz, C. J.

Seung, H. S.

Simon, B.

M. Debailleul, B. Simon, V. Georges, O. Haeberle, and V. Lauer, Meas. Sci. Technol. 19, 074009 (2008).
[CrossRef]

Song, S.

Stemmer, A.

Sung, Y.

B. Khaykovich, N. Kozlova, W. Choi, A. Lomakin, C. Hossain, Y. Sung, R. R. Dasari, M. S. Feld, and G. B. Benedek, Proc. Natl. Acad. Sci. USA 106, 15663 (2009).
[CrossRef] [PubMed]

Tada, Y.

Wicker, K.

Appl. Opt. (1)

Meas. Sci. Technol. (1)

M. Debailleul, B. Simon, V. Georges, O. Haeberle, and V. Lauer, Meas. Sci. Technol. 19, 074009 (2008).
[CrossRef]

Nat. Methods (1)

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, Nat. Methods 4, 717 (2007).
[CrossRef] [PubMed]

Opt. Express (2)

Opt. Lett. (3)

Proc. Natl. Acad. Sci. USA (1)

B. Khaykovich, N. Kozlova, W. Choi, A. Lomakin, C. Hossain, Y. Sung, R. R. Dasari, M. S. Feld, and G. B. Benedek, Proc. Natl. Acad. Sci. USA 106, 15663 (2009).
[CrossRef] [PubMed]

Other (1)

J. W. Goodman, Introduction to Fourier Optics (Roberts, 2005).

Supplementary Material (2)

» Media 1: MOV (1488 KB)     
» Media 2: MOV (2272 KB)     

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Figures (3)

Fig. 1
Fig. 1

High-speed synthetic aperture phase microscopy setup. Laser: He–Ne laser; GM, dual-axis galvanometer scanning mirror; L1, f = 250 mm lens; BF, backfocal plane of condenser lens; C, condenser lens; OL, objective lens; L2, tube lens; SF, spatial filter system. Inset, spiral trajectory of the focused spot at BF.

Fig. 2
Fig. 2

Angular images and synthesized images for a live microglia cell. (a), (d) Phase images taken in the image plane with illumination angles of 0 ° and 23.3 ° , respectively. (b) Phase image after background subtraction. (c) Phase image after aperture synthesis. Media 1 shows the dynamics of ruffles taken by the synthetic aperture quantitative phase microscopy. It takes one-fifth of a second to record one synthesized image, which is composed of 100 angle-dependent images taken at 500 frames per second. The cell is observed for 10 min. (e), (f) Intensity distribution in the Fourier plane for 0 ° illumination and for aperture synthesis, respectively. Color bars indicate phase in radians, and intensity in the logarithmic scale with base 10 for (e) and (f). Scale bar, 10 μm .

Fig. 3
Fig. 3

Images of a helical cholesterol ribbon [(a), (b)] and a USAF targetlike phase pattern [(c), (d)]. Phase images for angle of 0 ° illumination [(a), (c)] and for aperture synthesis [(b), (d)]. Media 2 is a movie showing the comparison between individual angular images on the left (panel a) and the synthesized image on the right (panel b). Color bars indicate phase in radians. Scale bar, 10 μm .

Equations (1)

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U f ( ξ , η ) = 1 i λ f circ ( ξ 2 + η 2 Δ ν ) T ( ξ / λ f + ν x , η / λ f + ν y ) .

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