Abstract

In this paper, we discuss a new methodology based on lensfree imaging to perform wound healing assay with unprecedented statistics. Our video lensfree microscopy setup is a simple device featuring only a CMOS sensor and a semi coherent illumination system. Yet it is a powerful mean for the real-time monitoring of cultivated cells. It presents several key advantages, e.g. integration into standard incubator, compatibility with standard cell culture protocol, simplicity and ease of use. It can perform the follow-up in a large field of view (25 mm2) of several crucial parameters during the culture of cells i.e. their motility, their proliferation rate or their death. Consequently the setup can gather large statistics both in space and time. Here we uses this facility in the context of wound healing assay to perform label-free measurements of the velocities of the fronts of proliferation of the cell layer as a function of time by means of particle image velocimetry (PIV) processing. However, for such tissue growth experiments, the field of view of 25 mm2 remains not sufficient and results can be biased depending on the position of the device with respect to the recipient of the cell culture. Hence, to conduct exhaustive wound healing assays, we propose to enlarge the field of view up to 10 cm2 through a raster scan, by moving the source/sensor with respect to the Petri dish. We have performed acquisitions of wound healing assay (keratinocytes HaCaT) both in real-time (25 mm2) and in final point (10 cm2) to assess the combination of velocimetry measurements and final point wide field imaging. In the future, we aim at combining directly our extended field of view acquisitions (>10 cm2) with real time ability inside the incubator.

© 2016 Optical Society of America

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  1. C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
    [Crossref]
  2. S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).
  3. S. Preibisch, S. Saalfeld, and P. Tomancak, “Globally optimal stitching of tiled 3D microscopic image acquisitions,” Bioinformatics 25(11), 1463–1465 (2009).
    [Crossref] [PubMed]
  4. J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
    [Crossref] [PubMed]
  5. C. A. Schneider, W. S. Rasband, and K. W. Eliceiri, “NIH Image to ImageJ: 25 years of image analysis,” Nature Methods 9, 671–675 (2012).
    [Crossref] [PubMed]
  6. M. Poujade, E. Grasland-Mongrain, A. Hertzog, J. Jouanneau, P. Chavrier, B. Ladoux, A. Buguin, and P. Silberzan, “Collective migration of an epithelial monolayer in response to a model wound,” Proc. Natl. Acad. Sci. USA 104(41), 15988–15993 (2007).
    [Crossref] [PubMed]
  7. M. Deforet, M. C. Parrini, L. Petitjean, M. Biondini, A. Buguin, J. Camonis, and P. Silberzan, “Automated velocity mapping of migrating cell populations (AVeMap),” Nature Methods 9, 1081–1083 (2012).
    [Crossref] [PubMed]
  8. T. W. Su, A. Erlinger, and A. Ozcan, “High-throughput lensfree imaging and characterization of a heterogeneous cell solution on a chip,” Biotechnol. Bioeng. 102(3), 856–868. (2009).
    [Crossref]
  9. S. O. Isikman, W. Bishara, S. Mavandadi, F. W. Yu, S. Feng, R. Lau, and A. Ozcan, “Lens-free optical tomographic microscope with a large imaging volume on a chip,” Proc. Natl. Acad. Sci. USA 108(18), 7296–7301. (2011).
    [Crossref] [PubMed]
  10. S. O. Isikman, A. Greenbaum, W. Luo, A. Coskun, and A. Ozcan, “Giga-pixel lensfree holographic microscopy and tomography using color image sensors,” PLoS ONE 7(9), e45044 (2012).
    [Crossref] [PubMed]
  11. A. Greenbaum, W. Luo, T. W. Su, Z. Grcs, L. Xue, S. O. Isikman, A. F. Coskun, O Mudanyali, and A. Ozcan, “Imaging without lenses: achievements and remaining challenges of wide-field on-chip microscopy,” Nature Methods 9(9), 889–895. (2012).
    [Crossref] [PubMed]

2014 (2)

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).

2012 (5)

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

C. A. Schneider, W. S. Rasband, and K. W. Eliceiri, “NIH Image to ImageJ: 25 years of image analysis,” Nature Methods 9, 671–675 (2012).
[Crossref] [PubMed]

M. Deforet, M. C. Parrini, L. Petitjean, M. Biondini, A. Buguin, J. Camonis, and P. Silberzan, “Automated velocity mapping of migrating cell populations (AVeMap),” Nature Methods 9, 1081–1083 (2012).
[Crossref] [PubMed]

S. O. Isikman, A. Greenbaum, W. Luo, A. Coskun, and A. Ozcan, “Giga-pixel lensfree holographic microscopy and tomography using color image sensors,” PLoS ONE 7(9), e45044 (2012).
[Crossref] [PubMed]

A. Greenbaum, W. Luo, T. W. Su, Z. Grcs, L. Xue, S. O. Isikman, A. F. Coskun, O Mudanyali, and A. Ozcan, “Imaging without lenses: achievements and remaining challenges of wide-field on-chip microscopy,” Nature Methods 9(9), 889–895. (2012).
[Crossref] [PubMed]

2011 (1)

S. O. Isikman, W. Bishara, S. Mavandadi, F. W. Yu, S. Feng, R. Lau, and A. Ozcan, “Lens-free optical tomographic microscope with a large imaging volume on a chip,” Proc. Natl. Acad. Sci. USA 108(18), 7296–7301. (2011).
[Crossref] [PubMed]

2009 (2)

T. W. Su, A. Erlinger, and A. Ozcan, “High-throughput lensfree imaging and characterization of a heterogeneous cell solution on a chip,” Biotechnol. Bioeng. 102(3), 856–868. (2009).
[Crossref]

S. Preibisch, S. Saalfeld, and P. Tomancak, “Globally optimal stitching of tiled 3D microscopic image acquisitions,” Bioinformatics 25(11), 1463–1465 (2009).
[Crossref] [PubMed]

2007 (1)

M. Poujade, E. Grasland-Mongrain, A. Hertzog, J. Jouanneau, P. Chavrier, B. Ladoux, A. Buguin, and P. Silberzan, “Collective migration of an epithelial monolayer in response to a model wound,” Proc. Natl. Acad. Sci. USA 104(41), 15988–15993 (2007).
[Crossref] [PubMed]

Allier, C.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).

Arganda-Carreras, I.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

Biondini, M.

M. Deforet, M. C. Parrini, L. Petitjean, M. Biondini, A. Buguin, J. Camonis, and P. Silberzan, “Automated velocity mapping of migrating cell populations (AVeMap),” Nature Methods 9, 1081–1083 (2012).
[Crossref] [PubMed]

Bishara, W.

S. O. Isikman, W. Bishara, S. Mavandadi, F. W. Yu, S. Feng, R. Lau, and A. Ozcan, “Lens-free optical tomographic microscope with a large imaging volume on a chip,” Proc. Natl. Acad. Sci. USA 108(18), 7296–7301. (2011).
[Crossref] [PubMed]

Buguin, A.

M. Deforet, M. C. Parrini, L. Petitjean, M. Biondini, A. Buguin, J. Camonis, and P. Silberzan, “Automated velocity mapping of migrating cell populations (AVeMap),” Nature Methods 9, 1081–1083 (2012).
[Crossref] [PubMed]

M. Poujade, E. Grasland-Mongrain, A. Hertzog, J. Jouanneau, P. Chavrier, B. Ladoux, A. Buguin, and P. Silberzan, “Collective migration of an epithelial monolayer in response to a model wound,” Proc. Natl. Acad. Sci. USA 104(41), 15988–15993 (2007).
[Crossref] [PubMed]

Camonis, J.

M. Deforet, M. C. Parrini, L. Petitjean, M. Biondini, A. Buguin, J. Camonis, and P. Silberzan, “Automated velocity mapping of migrating cell populations (AVeMap),” Nature Methods 9, 1081–1083 (2012).
[Crossref] [PubMed]

Cardona, A.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

Chalmond, B.

S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Chavrier, P.

M. Poujade, E. Grasland-Mongrain, A. Hertzog, J. Jouanneau, P. Chavrier, B. Ladoux, A. Buguin, and P. Silberzan, “Collective migration of an epithelial monolayer in response to a model wound,” Proc. Natl. Acad. Sci. USA 104(41), 15988–15993 (2007).
[Crossref] [PubMed]

Cioni, O.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Coskun, A.

S. O. Isikman, A. Greenbaum, W. Luo, A. Coskun, and A. Ozcan, “Giga-pixel lensfree holographic microscopy and tomography using color image sensors,” PLoS ONE 7(9), e45044 (2012).
[Crossref] [PubMed]

Coskun, A. F.

A. Greenbaum, W. Luo, T. W. Su, Z. Grcs, L. Xue, S. O. Isikman, A. F. Coskun, O Mudanyali, and A. Ozcan, “Imaging without lenses: achievements and remaining challenges of wide-field on-chip microscopy,” Nature Methods 9(9), 889–895. (2012).
[Crossref] [PubMed]

Coutard, J.-G.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

David-Watine, B.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).

Deforet, M.

M. Deforet, M. C. Parrini, L. Petitjean, M. Biondini, A. Buguin, J. Camonis, and P. Silberzan, “Automated velocity mapping of migrating cell populations (AVeMap),” Nature Methods 9, 1081–1083 (2012).
[Crossref] [PubMed]

Di Natale, C.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Dinten, J.-M.

S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Dolega, M. E.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Dubrulle, N.

S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).

Eliceiri, K. W.

C. A. Schneider, W. S. Rasband, and K. W. Eliceiri, “NIH Image to ImageJ: 25 years of image analysis,” Nature Methods 9, 671–675 (2012).
[Crossref] [PubMed]

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

Erlinger, A.

T. W. Su, A. Erlinger, and A. Ozcan, “High-throughput lensfree imaging and characterization of a heterogeneous cell solution on a chip,” Biotechnol. Bioeng. 102(3), 856–868. (2009).
[Crossref]

Feng, S.

S. O. Isikman, W. Bishara, S. Mavandadi, F. W. Yu, S. Feng, R. Lau, and A. Ozcan, “Lens-free optical tomographic microscope with a large imaging volume on a chip,” Proc. Natl. Acad. Sci. USA 108(18), 7296–7301. (2011).
[Crossref] [PubMed]

Freida, D.

S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).

Frise, E.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

Gidrol, X.

S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Grasland-Mongrain, E.

M. Poujade, E. Grasland-Mongrain, A. Hertzog, J. Jouanneau, P. Chavrier, B. Ladoux, A. Buguin, and P. Silberzan, “Collective migration of an epithelial monolayer in response to a model wound,” Proc. Natl. Acad. Sci. USA 104(41), 15988–15993 (2007).
[Crossref] [PubMed]

Grcs, Z.

A. Greenbaum, W. Luo, T. W. Su, Z. Grcs, L. Xue, S. O. Isikman, A. F. Coskun, O Mudanyali, and A. Ozcan, “Imaging without lenses: achievements and remaining challenges of wide-field on-chip microscopy,” Nature Methods 9(9), 889–895. (2012).
[Crossref] [PubMed]

Greenbaum, A.

A. Greenbaum, W. Luo, T. W. Su, Z. Grcs, L. Xue, S. O. Isikman, A. F. Coskun, O Mudanyali, and A. Ozcan, “Imaging without lenses: achievements and remaining challenges of wide-field on-chip microscopy,” Nature Methods 9(9), 889–895. (2012).
[Crossref] [PubMed]

S. O. Isikman, A. Greenbaum, W. Luo, A. Coskun, and A. Ozcan, “Giga-pixel lensfree holographic microscopy and tomography using color image sensors,” PLoS ONE 7(9), e45044 (2012).
[Crossref] [PubMed]

Haguet, V.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Hamard, L.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Hartenstein, V.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

Hertzog, A.

M. Poujade, E. Grasland-Mongrain, A. Hertzog, J. Jouanneau, P. Chavrier, B. Ladoux, A. Buguin, and P. Silberzan, “Collective migration of an epithelial monolayer in response to a model wound,” Proc. Natl. Acad. Sci. USA 104(41), 15988–15993 (2007).
[Crossref] [PubMed]

Isikman, S. O.

S. O. Isikman, A. Greenbaum, W. Luo, A. Coskun, and A. Ozcan, “Giga-pixel lensfree holographic microscopy and tomography using color image sensors,” PLoS ONE 7(9), e45044 (2012).
[Crossref] [PubMed]

A. Greenbaum, W. Luo, T. W. Su, Z. Grcs, L. Xue, S. O. Isikman, A. F. Coskun, O Mudanyali, and A. Ozcan, “Imaging without lenses: achievements and remaining challenges of wide-field on-chip microscopy,” Nature Methods 9(9), 889–895. (2012).
[Crossref] [PubMed]

S. O. Isikman, W. Bishara, S. Mavandadi, F. W. Yu, S. Feng, R. Lau, and A. Ozcan, “Lens-free optical tomographic microscope with a large imaging volume on a chip,” Proc. Natl. Acad. Sci. USA 108(18), 7296–7301. (2011).
[Crossref] [PubMed]

Jouanneau, J.

M. Poujade, E. Grasland-Mongrain, A. Hertzog, J. Jouanneau, P. Chavrier, B. Ladoux, A. Buguin, and P. Silberzan, “Collective migration of an epithelial monolayer in response to a model wound,” Proc. Natl. Acad. Sci. USA 104(41), 15988–15993 (2007).
[Crossref] [PubMed]

Kaynig, V.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

Kesavan, S. V.

S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Ladoux, B.

M. Poujade, E. Grasland-Mongrain, A. Hertzog, J. Jouanneau, P. Chavrier, B. Ladoux, A. Buguin, and P. Silberzan, “Collective migration of an epithelial monolayer in response to a model wound,” Proc. Natl. Acad. Sci. USA 104(41), 15988–15993 (2007).
[Crossref] [PubMed]

Lau, R.

S. O. Isikman, W. Bishara, S. Mavandadi, F. W. Yu, S. Feng, R. Lau, and A. Ozcan, “Lens-free optical tomographic microscope with a large imaging volume on a chip,” Proc. Natl. Acad. Sci. USA 108(18), 7296–7301. (2011).
[Crossref] [PubMed]

Longair, M.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

Luo, W.

S. O. Isikman, A. Greenbaum, W. Luo, A. Coskun, and A. Ozcan, “Giga-pixel lensfree holographic microscopy and tomography using color image sensors,” PLoS ONE 7(9), e45044 (2012).
[Crossref] [PubMed]

A. Greenbaum, W. Luo, T. W. Su, Z. Grcs, L. Xue, S. O. Isikman, A. F. Coskun, O Mudanyali, and A. Ozcan, “Imaging without lenses: achievements and remaining challenges of wide-field on-chip microscopy,” Nature Methods 9(9), 889–895. (2012).
[Crossref] [PubMed]

Mavandadi, S.

S. O. Isikman, W. Bishara, S. Mavandadi, F. W. Yu, S. Feng, R. Lau, and A. Ozcan, “Lens-free optical tomographic microscope with a large imaging volume on a chip,” Proc. Natl. Acad. Sci. USA 108(18), 7296–7301. (2011).
[Crossref] [PubMed]

Menneteau, M.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Momey, F.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).

Mudanyali, O

A. Greenbaum, W. Luo, T. W. Su, Z. Grcs, L. Xue, S. O. Isikman, A. F. Coskun, O Mudanyali, and A. Ozcan, “Imaging without lenses: achievements and remaining challenges of wide-field on-chip microscopy,” Nature Methods 9(9), 889–895. (2012).
[Crossref] [PubMed]

Navarro, F.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Obeid, P.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Ozcan, A.

A. Greenbaum, W. Luo, T. W. Su, Z. Grcs, L. Xue, S. O. Isikman, A. F. Coskun, O Mudanyali, and A. Ozcan, “Imaging without lenses: achievements and remaining challenges of wide-field on-chip microscopy,” Nature Methods 9(9), 889–895. (2012).
[Crossref] [PubMed]

S. O. Isikman, A. Greenbaum, W. Luo, A. Coskun, and A. Ozcan, “Giga-pixel lensfree holographic microscopy and tomography using color image sensors,” PLoS ONE 7(9), e45044 (2012).
[Crossref] [PubMed]

S. O. Isikman, W. Bishara, S. Mavandadi, F. W. Yu, S. Feng, R. Lau, and A. Ozcan, “Lens-free optical tomographic microscope with a large imaging volume on a chip,” Proc. Natl. Acad. Sci. USA 108(18), 7296–7301. (2011).
[Crossref] [PubMed]

T. W. Su, A. Erlinger, and A. Ozcan, “High-throughput lensfree imaging and characterization of a heterogeneous cell solution on a chip,” Biotechnol. Bioeng. 102(3), 856–868. (2009).
[Crossref]

Parrini, M. C.

M. Deforet, M. C. Parrini, L. Petitjean, M. Biondini, A. Buguin, J. Camonis, and P. Silberzan, “Automated velocity mapping of migrating cell populations (AVeMap),” Nature Methods 9, 1081–1083 (2012).
[Crossref] [PubMed]

Petitjean, L.

M. Deforet, M. C. Parrini, L. Petitjean, M. Biondini, A. Buguin, J. Camonis, and P. Silberzan, “Automated velocity mapping of migrating cell populations (AVeMap),” Nature Methods 9, 1081–1083 (2012).
[Crossref] [PubMed]

Picollet-D’hahan, N.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Pietzsch, T.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

Poujade, M.

M. Poujade, E. Grasland-Mongrain, A. Hertzog, J. Jouanneau, P. Chavrier, B. Ladoux, A. Buguin, and P. Silberzan, “Collective migration of an epithelial monolayer in response to a model wound,” Proc. Natl. Acad. Sci. USA 104(41), 15988–15993 (2007).
[Crossref] [PubMed]

Preibisch, S.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

S. Preibisch, S. Saalfeld, and P. Tomancak, “Globally optimal stitching of tiled 3D microscopic image acquisitions,” Bioinformatics 25(11), 1463–1465 (2009).
[Crossref] [PubMed]

Rasband, W. S.

C. A. Schneider, W. S. Rasband, and K. W. Eliceiri, “NIH Image to ImageJ: 25 years of image analysis,” Nature Methods 9, 671–675 (2012).
[Crossref] [PubMed]

Rueden, C.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

Saalfeld, S.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

S. Preibisch, S. Saalfeld, and P. Tomancak, “Globally optimal stitching of tiled 3D microscopic image acquisitions,” Bioinformatics 25(11), 1463–1465 (2009).
[Crossref] [PubMed]

Schindelin, J.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

Schmid, B.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

Schneider, C. A.

C. A. Schneider, W. S. Rasband, and K. W. Eliceiri, “NIH Image to ImageJ: 25 years of image analysis,” Nature Methods 9, 671–675 (2012).
[Crossref] [PubMed]

Shorte, S.

S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Silberzan, P.

M. Deforet, M. C. Parrini, L. Petitjean, M. Biondini, A. Buguin, J. Camonis, and P. Silberzan, “Automated velocity mapping of migrating cell populations (AVeMap),” Nature Methods 9, 1081–1083 (2012).
[Crossref] [PubMed]

M. Poujade, E. Grasland-Mongrain, A. Hertzog, J. Jouanneau, P. Chavrier, B. Ladoux, A. Buguin, and P. Silberzan, “Collective migration of an epithelial monolayer in response to a model wound,” Proc. Natl. Acad. Sci. USA 104(41), 15988–15993 (2007).
[Crossref] [PubMed]

Su, T. W.

A. Greenbaum, W. Luo, T. W. Su, Z. Grcs, L. Xue, S. O. Isikman, A. F. Coskun, O Mudanyali, and A. Ozcan, “Imaging without lenses: achievements and remaining challenges of wide-field on-chip microscopy,” Nature Methods 9(9), 889–895. (2012).
[Crossref] [PubMed]

T. W. Su, A. Erlinger, and A. Ozcan, “High-throughput lensfree imaging and characterization of a heterogeneous cell solution on a chip,” Biotechnol. Bioeng. 102(3), 856–868. (2009).
[Crossref]

Sulpice, E.

S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).

Tinevez, J.-Y.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

Tomancak, P.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

S. Preibisch, S. Saalfeld, and P. Tomancak, “Globally optimal stitching of tiled 3D microscopic image acquisitions,” Bioinformatics 25(11), 1463–1465 (2009).
[Crossref] [PubMed]

Van Der Sanden, B.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

White, D. J.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

Wion, D.

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Xue, L.

A. Greenbaum, W. Luo, T. W. Su, Z. Grcs, L. Xue, S. O. Isikman, A. F. Coskun, O Mudanyali, and A. Ozcan, “Imaging without lenses: achievements and remaining challenges of wide-field on-chip microscopy,” Nature Methods 9(9), 889–895. (2012).
[Crossref] [PubMed]

Yu, F. W.

S. O. Isikman, W. Bishara, S. Mavandadi, F. W. Yu, S. Feng, R. Lau, and A. Ozcan, “Lens-free optical tomographic microscope with a large imaging volume on a chip,” Proc. Natl. Acad. Sci. USA 108(18), 7296–7301. (2011).
[Crossref] [PubMed]

Bioinformatics (1)

S. Preibisch, S. Saalfeld, and P. Tomancak, “Globally optimal stitching of tiled 3D microscopic image acquisitions,” Bioinformatics 25(11), 1463–1465 (2009).
[Crossref] [PubMed]

Biotechnol. Bioeng. (1)

T. W. Su, A. Erlinger, and A. Ozcan, “High-throughput lensfree imaging and characterization of a heterogeneous cell solution on a chip,” Biotechnol. Bioeng. 102(3), 856–868. (2009).
[Crossref]

Nature Methods (4)

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. W. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nature Methods 9(7), 676–682 (2012).
[Crossref] [PubMed]

C. A. Schneider, W. S. Rasband, and K. W. Eliceiri, “NIH Image to ImageJ: 25 years of image analysis,” Nature Methods 9, 671–675 (2012).
[Crossref] [PubMed]

M. Deforet, M. C. Parrini, L. Petitjean, M. Biondini, A. Buguin, J. Camonis, and P. Silberzan, “Automated velocity mapping of migrating cell populations (AVeMap),” Nature Methods 9, 1081–1083 (2012).
[Crossref] [PubMed]

A. Greenbaum, W. Luo, T. W. Su, Z. Grcs, L. Xue, S. O. Isikman, A. F. Coskun, O Mudanyali, and A. Ozcan, “Imaging without lenses: achievements and remaining challenges of wide-field on-chip microscopy,” Nature Methods 9(9), 889–895. (2012).
[Crossref] [PubMed]

Nature Scientific Reports (1)

S. V. Kesavan, F. Momey, B. David-Watine, N. Dubrulle, S. Shorte, E. Sulpice, D. Freida, B. Chalmond, J.-M. Dinten, X. Gidrol, and C. Allier, “High-throughput monitoring of major cell functions by means of lensfree video microscopy,” Nature Scientific Reports 45942 (2014).

PLoS ONE (1)

S. O. Isikman, A. Greenbaum, W. Luo, A. Coskun, and A. Ozcan, “Giga-pixel lensfree holographic microscopy and tomography using color image sensors,” PLoS ONE 7(9), e45044 (2012).
[Crossref] [PubMed]

Proc. Natl. Acad. Sci. USA (2)

S. O. Isikman, W. Bishara, S. Mavandadi, F. W. Yu, S. Feng, R. Lau, and A. Ozcan, “Lens-free optical tomographic microscope with a large imaging volume on a chip,” Proc. Natl. Acad. Sci. USA 108(18), 7296–7301. (2011).
[Crossref] [PubMed]

M. Poujade, E. Grasland-Mongrain, A. Hertzog, J. Jouanneau, P. Chavrier, B. Ladoux, A. Buguin, and P. Silberzan, “Collective migration of an epithelial monolayer in response to a model wound,” Proc. Natl. Acad. Sci. USA 104(41), 15988–15993 (2007).
[Crossref] [PubMed]

Proc. SPIE (1)

C. Allier, S. V. Kesavan, J.-G. Coutard, O. Cioni, F. Momey, F. Navarro, M. Menneteau, B. Chalmond, P. Obeid, V. Haguet, B. David-Watine, S. Shorte, B. Van Der Sanden, C. Di Natale, L. Hamard, D. Wion, M. E. Dolega, N. Picollet-D’hahan, X. Gidrol, and J.-M. Dinten, “Video lensfree microscopy 2D and 3D culture of cells,” Proc. SPIE 8947, 89471H (2014).
[Crossref]

Supplementary Material (5)

NameDescription
» Visualization 1: AVI (11381 KB)      Real-time PIV lensfree sequence
» Visualization 2: AVI (10875 KB)      Real-time PIV lensfree sequence
» Visualization 3: AVI (3788 KB)      Real-time PIV lensfree sequence
» Visualization 4: AVI (8000 KB)      Real-time PIV lensfree sequence
» Visualization 5: AVI (7728 KB)      Real-time PIV lensfree sequence

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Figures (9)

Fig. 1
Fig. 1 (a) Experimental setup of the lensfree imaging device1. A CMOS image sensor is used for image acquisition and a LED and a 150μm pinhole for illumination (5cm above the image sensor). (b) The CMOS image sensor is put in contact with the Petri dish and records the holographic patterns resulting from the interference between the partially coherent incident light and the light scattered by the cells. Lensfree imaging is compatible with an incubation environment and can image 25 mm2 of, e.g., a standard 35mm Petri dish. (c) The image shows the acquisition of approximately 8000 fibroblast cells acquired by means of the lensfree video microscope.
Fig. 2
Fig. 2 (a) HaCaT culture in a Petri dish with a disk-shaped depletion plot (20mm). (b) Removing of the depletion plot at confluence. (c) Starting t0 of the growth experiment with real-time monitoring inside the incubator.
Fig. 3
Fig. 3 (a) Real-time lensfree sequence of HaCaT cells’ growth. (b) Image segmentation to get fronts positions. (c) PIV measurements of the front of proliferation. (d) PIV sequence of the front of proliferation (see Visualization 1). The colors distinguish isolated fronts to visualize the progressive junctions of the cell layer. (e) Scatter plot of the distribution of velocities over time. The red curves corresponds to the mean velocity (solid) its standard deviation (dash). Blanks correspond to loss of acquisition. The colorbar identifies the concentration (in %) of calculated velocity vectors with a given speed value (ordinate axis in μm/h) at a given time (abscissa axis in hours).
Fig. 4
Fig. 4 Experimental setup of the lensfree scanner. The light source, a white light LED with optical fiber + a wavelength filter at 534 ± 42nm, and the CMOS sensor are mounted on 2 linear stages set on XY configuration. The sample holder remains static while the source/sensor couple is displaced by the XY stages following a snake-like trajectory in order to acquire the mosaic of images covering the entire area of interest.
Fig. 5
Fig. 5 (a) Scanning trajectory of a Petri dish containing a culture of HaCaT cells and the corresponding mosaic of acquired images. (b) Meta-image of the Petri dish recombined by the stitching plugin [3] of Fiji [4] (ImageJ [5]).
Fig. 6
Fig. 6 Real-time lensfree sequences of 4 experiments (a, c, e, g) of HaCaT cultures (see Visualization 2, Visualization 3, Visualization 4 and Visualization 5) and their respective scatter graphs of distributions of proliferation velocities over time (b, d, f, h) as measured with PIV methods. The red curves corresponds to the mean velocity (solid) ± its standard deviation (dash). Blanks correspond to loss of acquisition. The red arrows on the graphs identify dates corresponding to the addition of culture medium.
Fig. 7
Fig. 7 Label-free measurement of the area of proliferation on the lensfree scan of the entire Petri dish of the HaCaT culture. (a) Meta-image at t0+96 hours. (b) Zoom on a region of interest. (c) Gradient image. The clusters corresponds to the growth of cells ripped at the removing of depletion plot. (d) Crowns’ delimitations at t0 and t0+96 hours for areas’ measurements. (f) Colorized Petri dish for classical non label-free measurements of proliferation. (g) Statistical results (mean and standard deviation) of quantification of the area of proliferation on N = 14 cell cultures (blue points). (h) Statistical results (mean and standard deviation) of quantification of the mean shifting at 4 cardinal points on N = 10 cell cultures (orange points).
Fig. 8
Fig. 8 Lensfree video microscope prototypes.(a) First lensfree video microscope prototype with a non-biocompatible exterior casing (Delrin - Polyoxymethylene). (b) Second prototype of lensfree video microscope, with a fan for ventilation (red arrow mark). Achievable temporal resolution is 15 minutes. The sensor is switched off in between image acquisitions. (c) Third prototype with Peltier element (red arrow), and an achievable temporal resolution close to 10 seconds. The imaging sensor is always on. (d) Lensfree video microscope where Peltier element is replaced by a heat-sink (red arrow), and opening for cross ventilation. The sensor is switched off in between acquisitions. Achievable temporal resolution is 15 minutes. All prototypes measure ∼ 10cm in height.
Fig. 9
Fig. 9 Temperature increase and frequency of imaging. 4 temperature sensors were used for evaluation. 2 temperature sensors were placed inside the incubator at different points (lines green, red). The other 2 temperature sensors were placed inside the culture dish (line purple) and on top of the imaging sensor (line blue). (a,b,c) Temperature increase during image acquisitions is observed for temporal resolutions of 20 minutes, 10 minutes, and 5 minutes respectively. (d) Temperature stability of the lensfree video microscope, with integrated temperature control. It is to be noted that the temperature inside the culture dish placed on the system and the temperature inside the incubator do not vary.

Equations (2)

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C t = I t * I t + 1
C i j t = C t ( i , j ) = m = 1 N n = 1 N I t ( m , n ) I t + 1 ( i + m , j + n )

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