Abstract

We present a simple-to-align, highly-portable interferometer, which is able to capture wide-field, off-axis interference patterns from transparent samples under low-coherence illumination. This small-dimensions and low-cost device can be connected to the output of a transmission microscope illuminated by a low-coherence source and measure sub-nanometric optical thickness changes in a label-free manner. In contrast to our previously published design, the τ interferometer, the new design is able to fully operate in an off-axis holographic geometry, where the interference fringes have high spatial frequency, and the interference area is limited only by the coherence length of the source, and thus it enables to easily obtain high-quality quantitative images of static and dynamic samples. We present several applications for the new design including nondestructive optical testing of transparent microscopic elements with nanometric thickness and live-cell imaging.

©2013 Optical Society of America

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  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).
  2. B. Rappaz, A. Barbul, Y. Emery, R. Korenstein, C. Depeursinge, P. J. Magistretti, and P. Marquet, “Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer,” Cytometry A 73A(10), 895–903 (2008).
  3. B. Kemper, A. Bauwens, A. Vollmer, S. Ketelhut, P. Langehanenberg, J. Müthing, H. Karch, and G. von Bally, “Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy,” J. Biomed. Opt. 15(3), 036009 (2010).
  4. N. T. Shaked, L. L. Satterwhite, M. J. Telen, G. A. Truskey, and A. Wax, “Quantitative microscopy and nanoscopy of sickle red blood cells performed by wide field digital interferometry,” J. Biomed. Opt. 16(3), 030506 (2011).
  5. P. Girshovitz and N. T. Shaked, “Generalized cell morphological parameters based on interferometric phase microscopy and their application to cell life cycle characterization,” Biomed. Opt. Express 3(8), 1757–1773 (2012).
  6. S. Gawad, M. Giugliano, M. Heuschkel, B. Wessling, H. Markram, U. Schnakenberg, P. Renaud, and H. Morgan, “Substrate arrays of iridium oxide microelectrodes for in vitro neuronal interfacing,” Front Neuroeng 2, 1–7 (2009).
  7. M. C. Potcoava and M. K. Kim, “Fingerprint biometry applications of digital holography and low-coherence interferography,” Appl. Opt. 48(34), H9–H15 (2009).
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  10. P. Bon, G. Maucort, B. Wattellier, and S. Monneret, “Quadriwave lateral shearing interferometry for quantitative phase microscopy of living cells,” Opt. Express 17(15), 13080–13094 (2009).
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  14. Z. Monemhaghdoust, F. Montfort, Y. Emery, C. Depeursinge, and C. Moser, “Dual wavelength full field imaging in low coherence digital holographic microscopy,” Opt. Express 19(24), 24005–24022 (2011).
  15. Z. Wang, L. J. Millet, M. Mir, H. Ding, S. Unarunotai, J. A. Rogers, M. U. Gillette, and G. Popescu, “Spatial light interference microscopy (SLIM),” Opt. Express 19(2), 1016–1026 (2011).
  16. B. Bhaduri, H. Pham, M. Mir, and G. Popescu, “Diffraction phase microscopy with white light,” Opt. Lett. 37(6), 1094–1096 (2012).
  17. N. T. Shaked, Y. Zhu, N. Badie, N. Bursac, and A. Wax, “Reflective interferometric chamber for quantitative phase imaging of biological sample dynamics,” J. Biomed. Opt. 15(3), 030503 (2010).
  18. J. Jang, C. Y. Bae, J.-K. Park, and J. C. Ye, “Self-reference quantitative phase microscopy for microfluidic devices,” Opt. Lett. 35(4), 514–516 (2010).
  19. B. Kemper, A. Vollmer, C. E. Rommel, J. Schnekenburger, and G. von Bally, “Simplified approach for quantitative digital holographic phase contrast imaging of living cells,” J. Biomed. Opt. 16(2), 026014 (2011).
  20. N. T. Shaked, “Quantitative phase microscopy of biological samples using a portable interferometer,” Opt. Lett. 37(11), 2016–2018 (2012).
  21. N. T. Shaked, T. M. Newpher, M. D. Ehlers, and A. Wax, “Parallel on-axis holographic phase microscopy of biological cells and unicellular microorganism dynamics,” Appl. Opt. 49(15), 2872–2878 (2010).
  22. 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).
  23. L. Xue, J. Lai, S. Wang, and Z. Li, “Single-shot slightly-off-axis interferometry based Hilbert phase microscopy of red blood cells,” Biomed. Opt. Express 2(4), 987–995 (2011).
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  28. 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).
  29. I. Shock, A. Barbul, P. Girshovitz, U. Nevo, R. Korenstein, and N. T. Shaked, “Optical phase nanoscopy in red blood cells using low-coherence spectroscopy,” J. Biomed. Opt. 17(10), 101509 (2012).
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2012 (4)

2011 (6)

2010 (6)

B. Kemper, A. Bauwens, A. Vollmer, S. Ketelhut, P. Langehanenberg, J. Müthing, H. Karch, and G. von Bally, “Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy,” J. Biomed. Opt. 15(3), 036009 (2010).

V. Micó and J. García, “Common-path phase-shifting lensless holographic microscopy,” Opt. Lett. 35(23), 3919–3921 (2010).

N. T. Shaked, Y. Zhu, N. Badie, N. Bursac, and A. Wax, “Reflective interferometric chamber for quantitative phase imaging of biological sample dynamics,” J. Biomed. Opt. 15(3), 030503 (2010).

J. Jang, C. Y. Bae, J.-K. Park, and J. C. Ye, “Self-reference quantitative phase microscopy for microfluidic devices,” Opt. Lett. 35(4), 514–516 (2010).

P. Kolman and R. Chmelík, “Coherence-controlled holographic microscope,” Opt. Express 18(21), 21990–22003 (2010).

N. T. Shaked, T. M. Newpher, M. D. Ehlers, and A. Wax, “Parallel on-axis holographic phase microscopy of biological cells and unicellular microorganism dynamics,” Appl. Opt. 49(15), 2872–2878 (2010).

2009 (3)

2008 (3)

V. Mico, Z. Zalevsky, and J. García, “Common-path phase-shifting digital holographic microscopy: a way to quantitative phase imaging and superresolution,” Opt. Commun. 281(17), 4273–4281 (2008).

B. Rappaz, A. Barbul, Y. Emery, R. Korenstein, C. Depeursinge, P. J. Magistretti, and P. Marquet, “Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer,” Cytometry A 73A(10), 895–903 (2008).

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).

2005 (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).

2004 (1)

2001 (1)

S. Reyntjens and R. Puers, “A review of focused ion beam applications in microsystem technology,” J. Micromech. Microeng. 11(4), 287–300 (2001).

1999 (1)

R. Chmelík and Z. Harna, “Parallel-mode confocal microscope,” Opt. Eng. 38(10), 1635–1639 (1999).

Badie, N.

N. T. Shaked, Y. Zhu, N. Badie, N. Bursac, and A. Wax, “Reflective interferometric chamber for quantitative phase imaging of biological sample dynamics,” J. Biomed. Opt. 15(3), 030503 (2010).

Badizadegan, K.

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).

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).

Bae, C. Y.

Barbul, A.

I. Shock, A. Barbul, P. Girshovitz, U. Nevo, R. Korenstein, and N. T. Shaked, “Optical phase nanoscopy in red blood cells using low-coherence spectroscopy,” J. Biomed. Opt. 17(10), 101509 (2012).

B. Rappaz, A. Barbul, Y. Emery, R. Korenstein, C. Depeursinge, P. J. Magistretti, and P. Marquet, “Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer,” Cytometry A 73A(10), 895–903 (2008).

Bauwens, A.

B. Kemper, A. Bauwens, A. Vollmer, S. Ketelhut, P. Langehanenberg, J. Müthing, H. Karch, and G. von Bally, “Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy,” J. Biomed. Opt. 15(3), 036009 (2010).

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).

Bhaduri, B.

Bon, P.

Bursac, N.

N. T. Shaked, Y. Zhu, N. Badie, N. Bursac, and A. Wax, “Reflective interferometric chamber for quantitative phase imaging of biological sample dynamics,” J. Biomed. Opt. 15(3), 030503 (2010).

Chmelík, R.

P. Kolman and R. Chmelík, “Coherence-controlled holographic microscope,” Opt. Express 18(21), 21990–22003 (2010).

R. Chmelík and Z. Harna, “Parallel-mode confocal microscope,” Opt. Eng. 38(10), 1635–1639 (1999).

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).

Dasari, R. R.

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).

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).

Deflores, L. P.

Depeursinge, C.

Z. Monemhaghdoust, F. Montfort, Y. Emery, C. Depeursinge, and C. Moser, “Dual wavelength full field imaging in low coherence digital holographic microscopy,” Opt. Express 19(24), 24005–24022 (2011).

B. Rappaz, A. Barbul, Y. Emery, R. Korenstein, C. Depeursinge, P. J. Magistretti, and P. Marquet, “Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer,” Cytometry A 73A(10), 895–903 (2008).

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).

Ding, H.

Ehlers, M. D.

Emery, Y.

Z. Monemhaghdoust, F. Montfort, Y. Emery, C. Depeursinge, and C. Moser, “Dual wavelength full field imaging in low coherence digital holographic microscopy,” Opt. Express 19(24), 24005–24022 (2011).

B. Rappaz, A. Barbul, Y. Emery, R. Korenstein, C. Depeursinge, P. J. Magistretti, and P. Marquet, “Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer,” Cytometry A 73A(10), 895–903 (2008).

Feld, M. 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).

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).

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).

García, J.

V. Micó and J. García, “Common-path phase-shifting lensless holographic microscopy,” Opt. Lett. 35(23), 3919–3921 (2010).

V. Mico, Z. Zalevsky, and J. García, “Common-path phase-shifting digital holographic microscopy: a way to quantitative phase imaging and superresolution,” Opt. Commun. 281(17), 4273–4281 (2008).

Gawad, S.

S. Gawad, M. Giugliano, M. Heuschkel, B. Wessling, H. Markram, U. Schnakenberg, P. Renaud, and H. Morgan, “Substrate arrays of iridium oxide microelectrodes for in vitro neuronal interfacing,” Front Neuroeng 2, 1–7 (2009).

Gillette, M. U.

Girshovitz, P.

P. Girshovitz and N. T. Shaked, “Generalized cell morphological parameters based on interferometric phase microscopy and their application to cell life cycle characterization,” Biomed. Opt. Express 3(8), 1757–1773 (2012).

I. Shock, A. Barbul, P. Girshovitz, U. Nevo, R. Korenstein, and N. T. Shaked, “Optical phase nanoscopy in red blood cells using low-coherence spectroscopy,” J. Biomed. Opt. 17(10), 101509 (2012).

Giugliano, M.

S. Gawad, M. Giugliano, M. Heuschkel, B. Wessling, H. Markram, U. Schnakenberg, P. Renaud, and H. Morgan, “Substrate arrays of iridium oxide microelectrodes for in vitro neuronal interfacing,” Front Neuroeng 2, 1–7 (2009).

Harna, Z.

R. Chmelík and Z. Harna, “Parallel-mode confocal microscope,” Opt. Eng. 38(10), 1635–1639 (1999).

Heuschkel, M.

S. Gawad, M. Giugliano, M. Heuschkel, B. Wessling, H. Markram, U. Schnakenberg, P. Renaud, and H. Morgan, “Substrate arrays of iridium oxide microelectrodes for in vitro neuronal interfacing,” Front Neuroeng 2, 1–7 (2009).

Ikeda, T.

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).

Iwai, H.

Jang, J.

Karch, H.

B. Kemper, A. Bauwens, A. Vollmer, S. Ketelhut, P. Langehanenberg, J. Müthing, H. Karch, and G. von Bally, “Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy,” J. Biomed. Opt. 15(3), 036009 (2010).

Kemper, B.

B. Kemper, A. Vollmer, C. E. Rommel, J. Schnekenburger, and G. von Bally, “Simplified approach for quantitative digital holographic phase contrast imaging of living cells,” J. Biomed. Opt. 16(2), 026014 (2011).

B. Kemper, A. Bauwens, A. Vollmer, S. Ketelhut, P. Langehanenberg, J. Müthing, H. Karch, and G. von Bally, “Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy,” J. Biomed. Opt. 15(3), 036009 (2010).

Ketelhut, S.

B. Kemper, A. Bauwens, A. Vollmer, S. Ketelhut, P. Langehanenberg, J. Müthing, H. Karch, and G. von Bally, “Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy,” J. Biomed. Opt. 15(3), 036009 (2010).

Kim, M. K.

Kolman, P.

Korenstein, R.

I. Shock, A. Barbul, P. Girshovitz, U. Nevo, R. Korenstein, and N. T. Shaked, “Optical phase nanoscopy in red blood cells using low-coherence spectroscopy,” J. Biomed. Opt. 17(10), 101509 (2012).

B. Rappaz, A. Barbul, Y. Emery, R. Korenstein, C. Depeursinge, P. J. Magistretti, and P. Marquet, “Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer,” Cytometry A 73A(10), 895–903 (2008).

Lai, J.

Langehanenberg, P.

B. Kemper, A. Bauwens, A. Vollmer, S. Ketelhut, P. Langehanenberg, J. Müthing, H. Karch, and G. von Bally, “Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy,” J. Biomed. Opt. 15(3), 036009 (2010).

Lee, M.

Li, Z.

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).

Magistretti, P. J.

B. Rappaz, A. Barbul, Y. Emery, R. Korenstein, C. Depeursinge, P. J. Magistretti, and P. Marquet, “Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer,” Cytometry A 73A(10), 895–903 (2008).

Markram, H.

S. Gawad, M. Giugliano, M. Heuschkel, B. Wessling, H. Markram, U. Schnakenberg, P. Renaud, and H. Morgan, “Substrate arrays of iridium oxide microelectrodes for in vitro neuronal interfacing,” Front Neuroeng 2, 1–7 (2009).

Marquet, P.

B. Rappaz, A. Barbul, Y. Emery, R. Korenstein, C. Depeursinge, P. J. Magistretti, and P. Marquet, “Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer,” Cytometry A 73A(10), 895–903 (2008).

Maucort, G.

Mico, V.

V. Mico, Z. Zalevsky, and J. García, “Common-path phase-shifting digital holographic microscopy: a way to quantitative phase imaging and superresolution,” Opt. Commun. 281(17), 4273–4281 (2008).

Micó, V.

Millet, L. J.

Mir, M.

Monemhaghdoust, Z.

Monneret, S.

Montfort, F.

Morgan, H.

S. Gawad, M. Giugliano, M. Heuschkel, B. Wessling, H. Markram, U. Schnakenberg, P. Renaud, and H. Morgan, “Substrate arrays of iridium oxide microelectrodes for in vitro neuronal interfacing,” Front Neuroeng 2, 1–7 (2009).

Moser, C.

Müthing, J.

B. Kemper, A. Bauwens, A. Vollmer, S. Ketelhut, P. Langehanenberg, J. Müthing, H. Karch, and G. von Bally, “Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy,” J. Biomed. Opt. 15(3), 036009 (2010).

Nevo, U.

I. Shock, A. Barbul, P. Girshovitz, U. Nevo, R. Korenstein, and N. T. Shaked, “Optical phase nanoscopy in red blood cells using low-coherence spectroscopy,” J. Biomed. Opt. 17(10), 101509 (2012).

Newpher, T. M.

Ozcan, A.

Park, J.-K.

Park, Y. K.

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).

Pham, H.

Popescu, G.

B. Bhaduri, H. Pham, M. Mir, and G. Popescu, “Diffraction phase microscopy with white light,” Opt. Lett. 37(6), 1094–1096 (2012).

Z. Wang, L. J. Millet, M. Mir, H. Ding, S. Unarunotai, J. A. Rogers, M. U. Gillette, and G. Popescu, “Spatial light interference microscopy (SLIM),” Opt. Express 19(2), 1016–1026 (2011).

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).

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).

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).

Potcoava, M. C.

Puers, R.

S. Reyntjens and R. Puers, “A review of focused ion beam applications in microsystem technology,” J. Micromech. Microeng. 11(4), 287–300 (2001).

Rappaz, B.

B. Rappaz, A. Barbul, Y. Emery, R. Korenstein, C. Depeursinge, P. J. Magistretti, and P. Marquet, “Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer,” Cytometry A 73A(10), 895–903 (2008).

Renaud, P.

S. Gawad, M. Giugliano, M. Heuschkel, B. Wessling, H. Markram, U. Schnakenberg, P. Renaud, and H. Morgan, “Substrate arrays of iridium oxide microelectrodes for in vitro neuronal interfacing,” Front Neuroeng 2, 1–7 (2009).

Reyntjens, S.

S. Reyntjens and R. Puers, “A review of focused ion beam applications in microsystem technology,” J. Micromech. Microeng. 11(4), 287–300 (2001).

Rogers, J. A.

Rommel, C. E.

B. Kemper, A. Vollmer, C. E. Rommel, J. Schnekenburger, and G. von Bally, “Simplified approach for quantitative digital holographic phase contrast imaging of living cells,” J. Biomed. Opt. 16(2), 026014 (2011).

Satterwhite, L. L.

N. T. Shaked, L. L. Satterwhite, M. J. Telen, G. A. Truskey, and A. Wax, “Quantitative microscopy and nanoscopy of sickle red blood cells performed by wide field digital interferometry,” J. Biomed. Opt. 16(3), 030506 (2011).

Schnakenberg, U.

S. Gawad, M. Giugliano, M. Heuschkel, B. Wessling, H. Markram, U. Schnakenberg, P. Renaud, and H. Morgan, “Substrate arrays of iridium oxide microelectrodes for in vitro neuronal interfacing,” Front Neuroeng 2, 1–7 (2009).

Schnekenburger, J.

B. Kemper, A. Vollmer, C. E. Rommel, J. Schnekenburger, and G. von Bally, “Simplified approach for quantitative digital holographic phase contrast imaging of living cells,” J. Biomed. Opt. 16(2), 026014 (2011).

Shaked, N. T.

I. Shock, A. Barbul, P. Girshovitz, U. Nevo, R. Korenstein, and N. T. Shaked, “Optical phase nanoscopy in red blood cells using low-coherence spectroscopy,” J. Biomed. Opt. 17(10), 101509 (2012).

N. T. Shaked, “Quantitative phase microscopy of biological samples using a portable interferometer,” Opt. Lett. 37(11), 2016–2018 (2012).

P. Girshovitz and N. T. Shaked, “Generalized cell morphological parameters based on interferometric phase microscopy and their application to cell life cycle characterization,” Biomed. Opt. Express 3(8), 1757–1773 (2012).

N. T. Shaked, L. L. Satterwhite, M. J. Telen, G. A. Truskey, and A. Wax, “Quantitative microscopy and nanoscopy of sickle red blood cells performed by wide field digital interferometry,” J. Biomed. Opt. 16(3), 030506 (2011).

N. T. Shaked, Y. Zhu, N. Badie, N. Bursac, and A. Wax, “Reflective interferometric chamber for quantitative phase imaging of biological sample dynamics,” J. Biomed. Opt. 15(3), 030503 (2010).

N. T. Shaked, T. M. Newpher, M. D. Ehlers, and A. Wax, “Parallel on-axis holographic phase microscopy of biological cells and unicellular microorganism dynamics,” Appl. Opt. 49(15), 2872–2878 (2010).

Shock, I.

I. Shock, A. Barbul, P. Girshovitz, U. Nevo, R. Korenstein, and N. T. Shaked, “Optical phase nanoscopy in red blood cells using low-coherence spectroscopy,” J. Biomed. Opt. 17(10), 101509 (2012).

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).

Telen, M. J.

N. T. Shaked, L. L. Satterwhite, M. J. Telen, G. A. Truskey, and A. Wax, “Quantitative microscopy and nanoscopy of sickle red blood cells performed by wide field digital interferometry,” J. Biomed. Opt. 16(3), 030506 (2011).

Truskey, G. A.

N. T. Shaked, L. L. Satterwhite, M. J. Telen, G. A. Truskey, and A. Wax, “Quantitative microscopy and nanoscopy of sickle red blood cells performed by wide field digital interferometry,” J. Biomed. Opt. 16(3), 030506 (2011).

Unarunotai, S.

Vaughan, J. C.

Vollmer, A.

B. Kemper, A. Vollmer, C. E. Rommel, J. Schnekenburger, and G. von Bally, “Simplified approach for quantitative digital holographic phase contrast imaging of living cells,” J. Biomed. Opt. 16(2), 026014 (2011).

B. Kemper, A. Bauwens, A. Vollmer, S. Ketelhut, P. Langehanenberg, J. Müthing, H. Karch, and G. von Bally, “Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy,” J. Biomed. Opt. 15(3), 036009 (2010).

von Bally, G.

B. Kemper, A. Vollmer, C. E. Rommel, J. Schnekenburger, and G. von Bally, “Simplified approach for quantitative digital holographic phase contrast imaging of living cells,” J. Biomed. Opt. 16(2), 026014 (2011).

B. Kemper, A. Bauwens, A. Vollmer, S. Ketelhut, P. Langehanenberg, J. Müthing, H. Karch, and G. von Bally, “Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy,” J. Biomed. Opt. 15(3), 036009 (2010).

Wang, S.

Wang, Z.

Wattellier, B.

Wax, A.

N. T. Shaked, L. L. Satterwhite, M. J. Telen, G. A. Truskey, and A. Wax, “Quantitative microscopy and nanoscopy of sickle red blood cells performed by wide field digital interferometry,” J. Biomed. Opt. 16(3), 030506 (2011).

N. T. Shaked, Y. Zhu, N. Badie, N. Bursac, and A. Wax, “Reflective interferometric chamber for quantitative phase imaging of biological sample dynamics,” J. Biomed. Opt. 15(3), 030503 (2010).

N. T. Shaked, T. M. Newpher, M. D. Ehlers, and A. Wax, “Parallel on-axis holographic phase microscopy of biological cells and unicellular microorganism dynamics,” Appl. Opt. 49(15), 2872–2878 (2010).

Wessling, B.

S. Gawad, M. Giugliano, M. Heuschkel, B. Wessling, H. Markram, U. Schnakenberg, P. Renaud, and H. Morgan, “Substrate arrays of iridium oxide microelectrodes for in vitro neuronal interfacing,” Front Neuroeng 2, 1–7 (2009).

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Zhu, Y.

N. T. Shaked, Y. Zhu, N. Badie, N. Bursac, and A. Wax, “Reflective interferometric chamber for quantitative phase imaging of biological sample dynamics,” J. Biomed. Opt. 15(3), 030503 (2010).

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Cytometry A (1)

B. Rappaz, A. Barbul, Y. Emery, R. Korenstein, C. Depeursinge, P. J. Magistretti, and P. Marquet, “Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer,” Cytometry A 73A(10), 895–903 (2008).

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B. Kemper, A. Bauwens, A. Vollmer, S. Ketelhut, P. Langehanenberg, J. Müthing, H. Karch, and G. von Bally, “Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy,” J. Biomed. Opt. 15(3), 036009 (2010).

N. T. Shaked, L. L. Satterwhite, M. J. Telen, G. A. Truskey, and A. Wax, “Quantitative microscopy and nanoscopy of sickle red blood cells performed by wide field digital interferometry,” J. Biomed. Opt. 16(3), 030506 (2011).

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).

N. T. Shaked, Y. Zhu, N. Badie, N. Bursac, and A. Wax, “Reflective interferometric chamber for quantitative phase imaging of biological sample dynamics,” J. Biomed. Opt. 15(3), 030503 (2010).

B. Kemper, A. Vollmer, C. E. Rommel, J. Schnekenburger, and G. von Bally, “Simplified approach for quantitative digital holographic phase contrast imaging of living cells,” J. Biomed. Opt. 16(2), 026014 (2011).

I. Shock, A. Barbul, P. Girshovitz, U. Nevo, R. Korenstein, and N. T. Shaked, “Optical phase nanoscopy in red blood cells using low-coherence spectroscopy,” J. Biomed. Opt. 17(10), 101509 (2012).

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Supplementary Material (1)

» Media 1: MPEG (148 KB)     

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Figures (11)

Fig. 1
Fig. 1

Schematic system diagrams of: (a) the conventional τ interferometer [20]; (b) the off-axis τ interferometer. L1,L2 – lenses in a 4f configuration, BS – beam splitter, M1,M2 – mirrors, P – pinhole, RR – retro-reflector made of a two-mirror construction.

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Fig. 2
Fig. 2

Explanation for the retro-reflector (RR) operation using ray tracing of the sample and the reference beams in the off-axis τ interferometer, as it would be seen if they were on the same optical axis.

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Fig. 3
Fig. 3

The off-axis τ interferometer, connected in the output of an inverted microscope, which is illuminated by a tunable low-coherence source. MO – microscope objective, L0,L1,L2 – lenses, where L1 and L2 are in a 4f configuration, BS – beam splitter, M,M2 – mirrors, P – pinhole, RR – two-mirror retro-reflector. Inset: Wide-field off-axis interferogram of red blood cells obtained with the system, and its cross-section at the location indicated by the white line.

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Fig. 4
Fig. 4

OPD sensitivities in a dry sample: (a) Spatial sensitivity: OPD standard deviation across a single OPD map for each of the 150 OPD maps. (b) Temporal sensitivity: OPD standard deviation for each diffraction-limited spot across the 150 OPD maps.

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Fig. 5
Fig. 5

OPD maps of a volume phase holographic grating obtained under low-coherence illumination by: (a) the off-axis τ interferometer; and (b) the off-axis Mach-Zehnder interferometer.

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Fig. 6
Fig. 6

SEM image of an element similar to our first lithographed phase target.

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Fig. 7
Fig. 7

OPD maps of the first phase target created by FIB lithography, containing variable depths elements (see Fig. 6), as obtained using: (a) the off-axis τ interferometer with a low-coherence source; (b) the off-axis Mach-Zehnder interferometer with a low-coherence source; and (c) the off-axis Mach-Zehnder interferometer with a high-coherence source (HeNe laser).

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Fig. 8
Fig. 8

OPD maps of the second phase target created by FIB lithography, containing variable depth elements, as obtained using: (a) the off-axis τ interferometer with a low-coherence source; (b) the off-axis Mach-Zehnder interferometer with a low-coherence source; and (c) the off-axis Mach-Zehnder interferometer with a high-coherence source (HeNe laser).

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Fig. 9
Fig. 9

OPD and physical thickness maps of RBCs, obtained using a low-coherence source in: (a) the off-axis τ interferometer; and (b) the off-axis Mach-Zehnder interferometer. The standard deviation of the OPD and of the physical thickness maps for: (c) the off-axis τ interferometer; and (d) the off-axis Mach-Zehnder interferometer.

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Fig. 10
Fig. 10

Measurements of Blepharisma organism swimming in water using the off-axis τ interferometer, demonstrating the system capabilities for quantitative imaging of fast dynamics on relatively large field of view due to its true off-axis configuration, and as opposed to the conventional τ interferometer [20]. See video in Media 1.

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Fig. 11
Fig. 11

Schematics of the sample and immersion medium thicknesses and refractive indices.

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Equations (11)

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θ=arctan(Δy/f),
I=〈 | V s + V r | 2 〉= I s + I r + G +1 + G −1 .
G +1 = I s I r ×exp[ − |OP D total | l c (x,y) ]×exp[ −j 2π λ OP D total ]×exp[ −j 2π λ ysin(θ) ],
OP D s (x,y)=[ n ¯ s (x,y)− n m ]× h s (x,y),
n ¯ s (x,y)= 1 h s (x,y) ∫ 0 h s (x,y) n s (x,y,z)dz .
G +1 =〈 V s * (t)× V r (t+τ)〉= I s I r ×exp[ − | τ | τ c ]×exp[ −j2π c λ τ ],
t 1 = d c ;   t 2 = 1 c { [d− h m (x,y)]+[ h m (x,y)− h s (x,y)]× n m (x,y)+ h s (x,y)× n ¯ s (x,y) }; τ= t 2 − t 1 = 1 c { h s (x,y)×[ n ¯ s (x,y)− n m (x,y)]+ h m (x,y)×[ n m (x,y)−1] },
OP D total (x,y)=c⋅τ                               = h s (x,y)×[ n ¯ s (x,y)− n m (x,y)]+ h m (x,y)×[ n m (x,y)−1]                               =OP D s +OP D m  ,
G +1 =   I s I r ×exp[ −| OP D total c | c l c ]×exp[ −j2π c λ OP D total c ]          = I s I r ×exp[ −| OP D total l c | ]×exp[ −j 2π λ OP D total ].
l c
G +1 = I r I s exp[ − |OP D total | l c (x,y) ]×exp[ −j 2π λ OP D total ]×exp[ −j 2π λ ysin(θ) ],

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