Abstract

Digital holography enables a multifocus quantitative phase microscopy for the investigation of reflective surfaces and for marker-free live cell imaging. For digital holographic long-term investigations of living cells an automated (subsequent) robust and reliable numerical focus adjustment is of particular importance. Four numerical methods for the determination of the optimal focus position in the numerical reconstruction and propagation of the complex object waves of pure phase objects are characterized, compared, and adapted to the requirements of digital holographic microscopy. Results from investigations of an engineered surface and human pancreas tumor cells demonstrate the applicability of Fourier-weighting- and gradient-operator-based methods for robust and reliable automated subsequent numerical digital holographic focusing.

© 2008 Optical Society of America

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  1. E. Cuche, P. Marquet, and C. Depeursinge, “Simultaneous amplitude-contrast and quantitative phase-contrast microscopy by numerical reconstruction of Fresnel off-axis holograms,” Appl.Opt. 38, 6994-7001 (1999).
  2. P. Marquet, B. Rappaz, P. J. Magistretti, E. Cuche, Y. Emery, T. Colomb, and C. Depeursinge, “Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy,” Opt. Lett. 30, 468-470 (2005).
    [CrossRef]
  3. D. Carl, B. Kemper, G. Wernicke, and G. von Bally, “Parameter-optimized digital holographic microscope for high resolution living cell analysis,” Appl. Opt. 43, 6536-6544 (2004).
    [CrossRef]
  4. B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schäfer, W. Domschke, and G. von Bally, “Investigations on living pancreas tumor cells by digital holographic microscopy,” J. Biomed. Opt. 11034005 (2006).
  5. Y. Sun, S. Duthaler, and B. J. Nelson, “Autofocusing in computer microscopy: selecting the optimal focus algorithm,” Microsc. Res. Tech. 65, 139-149 (2004).
    [CrossRef]
  6. F. Wolf and S. Geley, “A simple and stable autofocusing protocol for long multidimensional live cell microscopy,” J. Microsc. 221, 72-77 (2006).
    [CrossRef]
  7. M. Liebling and M. Unser, “Autofocus for digital Fresnel holograms by use of a Fresnelet-sparsity criterion,” J. Opt. Soc. Am. A 21, 2424-2430 (2004).
  8. B. Kemper, D. Carl, A. Höink, G. von Bally, I. Bredebusch, and J. Schnekenburger, “Modular digital holographic microscopy system for marker-free quantitative phase contrast imaging of living cells,” Proc. SPIE 6191, 61910T (2006).
  9. T. Kreis, Handbook of Holographic Interferometry: Optical and Digital Methods (Wiley-VCH, 2005).
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    [CrossRef]
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    [CrossRef]
  13. F. C. Groen, I. T. Young, and G. Ligthart, “A comparison of different focus functions for use in autofocus algorithms,” Cytometry 6, 81-91 (1985).
    [CrossRef]
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    [CrossRef]
  15. M. Bravo-Zanoguera, B. v. Massenbach, A. L. Kellner, and J. H. Price, “High-performance autofocus circuit for biological microscopy,” Rev. Sci. Instrum. 69, 3966-3977 (1998).
    [CrossRef]
  16. M. T. Özgen and T. E. Tuncer, “Object reconstruction from in-line Fresnel holograms without explicit depth focusing,” Opt. Eng. 43, 1300-1310 (2004).
  17. J.-M. Geusebroek, F. Cornelissen, A. W. M. Smeulders, and H. Geerts, “Robust autofocusing in microscopy,” Cytometry 39, 1-9 (2000).
    [CrossRef]
  18. A. Thelen, J. Bongartz, D. Giel, S. Frey, and P. Hering, “Iterative focus detection in hologram tomography,” J. Opt. Soc. Am. A 22, 1176-1180 (2005).
    [CrossRef]
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  20. W. Li, N. C. Loomis, Q. Hu, and C. S. Davis, “Focus detection from digital in-line holograms based on spectral l1 norms,” J. Opt. Soc. Am. A 24, 3054-3062 (2007).
    [CrossRef]

2007

2006

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schäfer, W. Domschke, and G. von Bally, “Investigations on living pancreas tumor cells by digital holographic microscopy,” J. Biomed. Opt. 11034005 (2006).

F. Wolf and S. Geley, “A simple and stable autofocusing protocol for long multidimensional live cell microscopy,” J. Microsc. 221, 72-77 (2006).
[CrossRef]

B. Kemper, D. Carl, A. Höink, G. von Bally, I. Bredebusch, and J. Schnekenburger, “Modular digital holographic microscopy system for marker-free quantitative phase contrast imaging of living cells,” Proc. SPIE 6191, 61910T (2006).

F. Dubois, C. Schockaert, N. Callens, and C. Yourassowsky, “Focus plane detection criteria in digital holography microscopy by amplitude analysis,” Opt. Express 14, 5895-5908(2006).
[CrossRef]

2005

2004

D. Carl, B. Kemper, G. Wernicke, and G. von Bally, “Parameter-optimized digital holographic microscope for high resolution living cell analysis,” Appl. Opt. 43, 6536-6544 (2004).
[CrossRef]

Y. Sun, S. Duthaler, and B. J. Nelson, “Autofocusing in computer microscopy: selecting the optimal focus algorithm,” Microsc. Res. Tech. 65, 139-149 (2004).
[CrossRef]

M. Liebling, T. Blu, and M. Unser, “Complex-wave retrieval from a single off-axis hologram,” J Opt Soc Am A 21, 367-377(2004).

M. Liebling and M. Unser, “Autofocus for digital Fresnel holograms by use of a Fresnelet-sparsity criterion,” J. Opt. Soc. Am. A 21, 2424-2430 (2004).

M. T. Özgen and T. E. Tuncer, “Object reconstruction from in-line Fresnel holograms without explicit depth focusing,” Opt. Eng. 43, 1300-1310 (2004).

2002

U. Schnars and W. Jüptner, “Digital recording and numerical reconstruction of holograms,” Meas. Sci. Technol. 13, R85-R101 (2002).
[CrossRef]

2000

J.-M. Geusebroek, F. Cornelissen, A. W. M. Smeulders, and H. Geerts, “Robust autofocusing in microscopy,” Cytometry 39, 1-9 (2000).
[CrossRef]

1999

E. Cuche, P. Marquet, and C. Depeursinge, “Simultaneous amplitude-contrast and quantitative phase-contrast microscopy by numerical reconstruction of Fresnel off-axis holograms,” Appl.Opt. 38, 6994-7001 (1999).

1998

M. Bravo-Zanoguera, B. v. Massenbach, A. L. Kellner, and J. H. Price, “High-performance autofocus circuit for biological microscopy,” Rev. Sci. Instrum. 69, 3966-3977 (1998).
[CrossRef]

1992

H. P. Elsässer, U. Lehr, B. Agricola, and H. F. Kern, “Establishment, and characterization of two cell lines with different grades of differentiation derived from one primary human pancreatic adenocarcinoma,” Virchows Arch. 61,295-306(1992).

1991

L. Firestone, K. Cook, K. Culp, N. Talsania, and K. Preston Jr, “Comparison of autofocus methods for automated microscopy,” Cytometry 12, 195-206 (1991).
[CrossRef]

1985

F. C. Groen, I. T. Young, and G. Ligthart, “A comparison of different focus functions for use in autofocus algorithms,” Cytometry 6, 81-91 (1985).
[CrossRef]

Agricola, B.

H. P. Elsässer, U. Lehr, B. Agricola, and H. F. Kern, “Establishment, and characterization of two cell lines with different grades of differentiation derived from one primary human pancreatic adenocarcinoma,” Virchows Arch. 61,295-306(1992).

Blu, T.

M. Liebling, T. Blu, and M. Unser, “Complex-wave retrieval from a single off-axis hologram,” J Opt Soc Am A 21, 367-377(2004).

Bongartz, J.

Bravo-Zanoguera, M.

M. Bravo-Zanoguera, B. v. Massenbach, A. L. Kellner, and J. H. Price, “High-performance autofocus circuit for biological microscopy,” Rev. Sci. Instrum. 69, 3966-3977 (1998).
[CrossRef]

Bredebusch, I.

B. Kemper, D. Carl, A. Höink, G. von Bally, I. Bredebusch, and J. Schnekenburger, “Modular digital holographic microscopy system for marker-free quantitative phase contrast imaging of living cells,” Proc. SPIE 6191, 61910T (2006).

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schäfer, W. Domschke, and G. von Bally, “Investigations on living pancreas tumor cells by digital holographic microscopy,” J. Biomed. Opt. 11034005 (2006).

Callens, N.

Carl, D.

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schäfer, W. Domschke, and G. von Bally, “Investigations on living pancreas tumor cells by digital holographic microscopy,” J. Biomed. Opt. 11034005 (2006).

B. Kemper, D. Carl, A. Höink, G. von Bally, I. Bredebusch, and J. Schnekenburger, “Modular digital holographic microscopy system for marker-free quantitative phase contrast imaging of living cells,” Proc. SPIE 6191, 61910T (2006).

D. Carl, B. Kemper, G. Wernicke, and G. von Bally, “Parameter-optimized digital holographic microscope for high resolution living cell analysis,” Appl. Opt. 43, 6536-6544 (2004).
[CrossRef]

Colomb, T.

Cook, K.

L. Firestone, K. Cook, K. Culp, N. Talsania, and K. Preston Jr, “Comparison of autofocus methods for automated microscopy,” Cytometry 12, 195-206 (1991).
[CrossRef]

Cornelissen, F.

J.-M. Geusebroek, F. Cornelissen, A. W. M. Smeulders, and H. Geerts, “Robust autofocusing in microscopy,” Cytometry 39, 1-9 (2000).
[CrossRef]

Cuche, E.

P. Marquet, B. Rappaz, P. J. Magistretti, E. Cuche, Y. Emery, T. Colomb, and C. Depeursinge, “Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy,” Opt. Lett. 30, 468-470 (2005).
[CrossRef]

E. Cuche, P. Marquet, and C. Depeursinge, “Simultaneous amplitude-contrast and quantitative phase-contrast microscopy by numerical reconstruction of Fresnel off-axis holograms,” Appl.Opt. 38, 6994-7001 (1999).

Culp, K.

L. Firestone, K. Cook, K. Culp, N. Talsania, and K. Preston Jr, “Comparison of autofocus methods for automated microscopy,” Cytometry 12, 195-206 (1991).
[CrossRef]

Davis, C. S.

Depeursinge, C.

P. Marquet, B. Rappaz, P. J. Magistretti, E. Cuche, Y. Emery, T. Colomb, and C. Depeursinge, “Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy,” Opt. Lett. 30, 468-470 (2005).
[CrossRef]

E. Cuche, P. Marquet, and C. Depeursinge, “Simultaneous amplitude-contrast and quantitative phase-contrast microscopy by numerical reconstruction of Fresnel off-axis holograms,” Appl.Opt. 38, 6994-7001 (1999).

Domschke, W.

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schäfer, W. Domschke, and G. von Bally, “Investigations on living pancreas tumor cells by digital holographic microscopy,” J. Biomed. Opt. 11034005 (2006).

Dubois, F.

Duthaler, S.

Y. Sun, S. Duthaler, and B. J. Nelson, “Autofocusing in computer microscopy: selecting the optimal focus algorithm,” Microsc. Res. Tech. 65, 139-149 (2004).
[CrossRef]

Elsässer, H. P.

H. P. Elsässer, U. Lehr, B. Agricola, and H. F. Kern, “Establishment, and characterization of two cell lines with different grades of differentiation derived from one primary human pancreatic adenocarcinoma,” Virchows Arch. 61,295-306(1992).

Emery, Y.

Firestone, L.

L. Firestone, K. Cook, K. Culp, N. Talsania, and K. Preston Jr, “Comparison of autofocus methods for automated microscopy,” Cytometry 12, 195-206 (1991).
[CrossRef]

Frey, S.

Geerts, H.

J.-M. Geusebroek, F. Cornelissen, A. W. M. Smeulders, and H. Geerts, “Robust autofocusing in microscopy,” Cytometry 39, 1-9 (2000).
[CrossRef]

Geley, S.

F. Wolf and S. Geley, “A simple and stable autofocusing protocol for long multidimensional live cell microscopy,” J. Microsc. 221, 72-77 (2006).
[CrossRef]

Geusebroek, J.-M.

J.-M. Geusebroek, F. Cornelissen, A. W. M. Smeulders, and H. Geerts, “Robust autofocusing in microscopy,” Cytometry 39, 1-9 (2000).
[CrossRef]

Giel, D.

Groen, F. C.

F. C. Groen, I. T. Young, and G. Ligthart, “A comparison of different focus functions for use in autofocus algorithms,” Cytometry 6, 81-91 (1985).
[CrossRef]

Hering, P.

Höink, A.

B. Kemper, D. Carl, A. Höink, G. von Bally, I. Bredebusch, and J. Schnekenburger, “Modular digital holographic microscopy system for marker-free quantitative phase contrast imaging of living cells,” Proc. SPIE 6191, 61910T (2006).

Hu, Q.

Jüptner, W.

U. Schnars and W. Jüptner, “Digital recording and numerical reconstruction of holograms,” Meas. Sci. Technol. 13, R85-R101 (2002).
[CrossRef]

Kellner, A. L.

M. Bravo-Zanoguera, B. v. Massenbach, A. L. Kellner, and J. H. Price, “High-performance autofocus circuit for biological microscopy,” Rev. Sci. Instrum. 69, 3966-3977 (1998).
[CrossRef]

Kemper, B.

B. Kemper, D. Carl, A. Höink, G. von Bally, I. Bredebusch, and J. Schnekenburger, “Modular digital holographic microscopy system for marker-free quantitative phase contrast imaging of living cells,” Proc. SPIE 6191, 61910T (2006).

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schäfer, W. Domschke, and G. von Bally, “Investigations on living pancreas tumor cells by digital holographic microscopy,” J. Biomed. Opt. 11034005 (2006).

D. Carl, B. Kemper, G. Wernicke, and G. von Bally, “Parameter-optimized digital holographic microscope for high resolution living cell analysis,” Appl. Opt. 43, 6536-6544 (2004).
[CrossRef]

Kern, H. F.

H. P. Elsässer, U. Lehr, B. Agricola, and H. F. Kern, “Establishment, and characterization of two cell lines with different grades of differentiation derived from one primary human pancreatic adenocarcinoma,” Virchows Arch. 61,295-306(1992).

Kreis, T.

T. Kreis, Handbook of Holographic Interferometry: Optical and Digital Methods (Wiley-VCH, 2005).

Lehr, U.

H. P. Elsässer, U. Lehr, B. Agricola, and H. F. Kern, “Establishment, and characterization of two cell lines with different grades of differentiation derived from one primary human pancreatic adenocarcinoma,” Virchows Arch. 61,295-306(1992).

Li, W.

Liebling, M.

M. Liebling and M. Unser, “Autofocus for digital Fresnel holograms by use of a Fresnelet-sparsity criterion,” J. Opt. Soc. Am. A 21, 2424-2430 (2004).

M. Liebling, T. Blu, and M. Unser, “Complex-wave retrieval from a single off-axis hologram,” J Opt Soc Am A 21, 367-377(2004).

Ligthart, G.

F. C. Groen, I. T. Young, and G. Ligthart, “A comparison of different focus functions for use in autofocus algorithms,” Cytometry 6, 81-91 (1985).
[CrossRef]

Loomis, N. C.

Magistretti, P. J.

Marquet, P.

P. Marquet, B. Rappaz, P. J. Magistretti, E. Cuche, Y. Emery, T. Colomb, and C. Depeursinge, “Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy,” Opt. Lett. 30, 468-470 (2005).
[CrossRef]

E. Cuche, P. Marquet, and C. Depeursinge, “Simultaneous amplitude-contrast and quantitative phase-contrast microscopy by numerical reconstruction of Fresnel off-axis holograms,” Appl.Opt. 38, 6994-7001 (1999).

Nelson, B. J.

Y. Sun, S. Duthaler, and B. J. Nelson, “Autofocusing in computer microscopy: selecting the optimal focus algorithm,” Microsc. Res. Tech. 65, 139-149 (2004).
[CrossRef]

Özgen, M. T.

M. T. Özgen and T. E. Tuncer, “Object reconstruction from in-line Fresnel holograms without explicit depth focusing,” Opt. Eng. 43, 1300-1310 (2004).

Preston, K.

L. Firestone, K. Cook, K. Culp, N. Talsania, and K. Preston Jr, “Comparison of autofocus methods for automated microscopy,” Cytometry 12, 195-206 (1991).
[CrossRef]

Price, J. H.

M. Bravo-Zanoguera, B. v. Massenbach, A. L. Kellner, and J. H. Price, “High-performance autofocus circuit for biological microscopy,” Rev. Sci. Instrum. 69, 3966-3977 (1998).
[CrossRef]

Rappaz, B.

Schäfer, M.

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schäfer, W. Domschke, and G. von Bally, “Investigations on living pancreas tumor cells by digital holographic microscopy,” J. Biomed. Opt. 11034005 (2006).

Schnars, U.

U. Schnars and W. Jüptner, “Digital recording and numerical reconstruction of holograms,” Meas. Sci. Technol. 13, R85-R101 (2002).
[CrossRef]

Schnekenburger, J.

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schäfer, W. Domschke, and G. von Bally, “Investigations on living pancreas tumor cells by digital holographic microscopy,” J. Biomed. Opt. 11034005 (2006).

B. Kemper, D. Carl, A. Höink, G. von Bally, I. Bredebusch, and J. Schnekenburger, “Modular digital holographic microscopy system for marker-free quantitative phase contrast imaging of living cells,” Proc. SPIE 6191, 61910T (2006).

Schockaert, C.

Smeulders, A. W. M.

J.-M. Geusebroek, F. Cornelissen, A. W. M. Smeulders, and H. Geerts, “Robust autofocusing in microscopy,” Cytometry 39, 1-9 (2000).
[CrossRef]

Sun, Y.

Y. Sun, S. Duthaler, and B. J. Nelson, “Autofocusing in computer microscopy: selecting the optimal focus algorithm,” Microsc. Res. Tech. 65, 139-149 (2004).
[CrossRef]

Talsania, N.

L. Firestone, K. Cook, K. Culp, N. Talsania, and K. Preston Jr, “Comparison of autofocus methods for automated microscopy,” Cytometry 12, 195-206 (1991).
[CrossRef]

Thelen, A.

Tuncer, T. E.

M. T. Özgen and T. E. Tuncer, “Object reconstruction from in-line Fresnel holograms without explicit depth focusing,” Opt. Eng. 43, 1300-1310 (2004).

Unser, M.

M. Liebling, T. Blu, and M. Unser, “Complex-wave retrieval from a single off-axis hologram,” J Opt Soc Am A 21, 367-377(2004).

M. Liebling and M. Unser, “Autofocus for digital Fresnel holograms by use of a Fresnelet-sparsity criterion,” J. Opt. Soc. Am. A 21, 2424-2430 (2004).

v. Massenbach, B.

M. Bravo-Zanoguera, B. v. Massenbach, A. L. Kellner, and J. H. Price, “High-performance autofocus circuit for biological microscopy,” Rev. Sci. Instrum. 69, 3966-3977 (1998).
[CrossRef]

von Bally, G.

B. Kemper, D. Carl, A. Höink, G. von Bally, I. Bredebusch, and J. Schnekenburger, “Modular digital holographic microscopy system for marker-free quantitative phase contrast imaging of living cells,” Proc. SPIE 6191, 61910T (2006).

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schäfer, W. Domschke, and G. von Bally, “Investigations on living pancreas tumor cells by digital holographic microscopy,” J. Biomed. Opt. 11034005 (2006).

D. Carl, B. Kemper, G. Wernicke, and G. von Bally, “Parameter-optimized digital holographic microscope for high resolution living cell analysis,” Appl. Opt. 43, 6536-6544 (2004).
[CrossRef]

Wernicke, G.

Wolf, F.

F. Wolf and S. Geley, “A simple and stable autofocusing protocol for long multidimensional live cell microscopy,” J. Microsc. 221, 72-77 (2006).
[CrossRef]

Young, I. T.

F. C. Groen, I. T. Young, and G. Ligthart, “A comparison of different focus functions for use in autofocus algorithms,” Cytometry 6, 81-91 (1985).
[CrossRef]

Yourassowsky, C.

Appl. Opt.

Appl.Opt.

E. Cuche, P. Marquet, and C. Depeursinge, “Simultaneous amplitude-contrast and quantitative phase-contrast microscopy by numerical reconstruction of Fresnel off-axis holograms,” Appl.Opt. 38, 6994-7001 (1999).

Cytometry

J.-M. Geusebroek, F. Cornelissen, A. W. M. Smeulders, and H. Geerts, “Robust autofocusing in microscopy,” Cytometry 39, 1-9 (2000).
[CrossRef]

F. C. Groen, I. T. Young, and G. Ligthart, “A comparison of different focus functions for use in autofocus algorithms,” Cytometry 6, 81-91 (1985).
[CrossRef]

L. Firestone, K. Cook, K. Culp, N. Talsania, and K. Preston Jr, “Comparison of autofocus methods for automated microscopy,” Cytometry 12, 195-206 (1991).
[CrossRef]

J Opt Soc Am A

M. Liebling, T. Blu, and M. Unser, “Complex-wave retrieval from a single off-axis hologram,” J Opt Soc Am A 21, 367-377(2004).

J. Biomed. Opt.

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schäfer, W. Domschke, and G. von Bally, “Investigations on living pancreas tumor cells by digital holographic microscopy,” J. Biomed. Opt. 11034005 (2006).

J. Microsc.

F. Wolf and S. Geley, “A simple and stable autofocusing protocol for long multidimensional live cell microscopy,” J. Microsc. 221, 72-77 (2006).
[CrossRef]

J. Opt. Soc. Am. A

Meas. Sci. Technol.

U. Schnars and W. Jüptner, “Digital recording and numerical reconstruction of holograms,” Meas. Sci. Technol. 13, R85-R101 (2002).
[CrossRef]

Microsc. Res. Tech.

Y. Sun, S. Duthaler, and B. J. Nelson, “Autofocusing in computer microscopy: selecting the optimal focus algorithm,” Microsc. Res. Tech. 65, 139-149 (2004).
[CrossRef]

Opt. Eng.

M. T. Özgen and T. E. Tuncer, “Object reconstruction from in-line Fresnel holograms without explicit depth focusing,” Opt. Eng. 43, 1300-1310 (2004).

Opt. Express

Opt. Lett.

Proc. SPIE

B. Kemper, D. Carl, A. Höink, G. von Bally, I. Bredebusch, and J. Schnekenburger, “Modular digital holographic microscopy system for marker-free quantitative phase contrast imaging of living cells,” Proc. SPIE 6191, 61910T (2006).

Rev. Sci. Instrum.

M. Bravo-Zanoguera, B. v. Massenbach, A. L. Kellner, and J. H. Price, “High-performance autofocus circuit for biological microscopy,” Rev. Sci. Instrum. 69, 3966-3977 (1998).
[CrossRef]

Virchows Arch.

H. P. Elsässer, U. Lehr, B. Agricola, and H. F. Kern, “Establishment, and characterization of two cell lines with different grades of differentiation derived from one primary human pancreatic adenocarcinoma,” Virchows Arch. 61,295-306(1992).

Other

T. Kreis, Handbook of Holographic Interferometry: Optical and Digital Methods (Wiley-VCH, 2005).

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