Abstract

A portable fluorescence optical detection system was developed to demonstrate real-time in situ analysis of cells that are three-dimensionally cultured in an extracellular matrix under microfluidic environment. The system was designed to provide a large field of view in the lateral plane to average out cellular processes in an axial layer and simultaneously diffraction-limited axial resolution. In this proof-of-concept study, the detection system was applied to quantitative analyses of short-term measurements of cell staining and cell cytotoxicity and long-term monitoring of a cell-invasion assay. For assays, colon cancer cells were cultured in a Matrigel or alginate matrix. The measured data were largely consistent with predicted results and revealed quantitatively cell dynamics specific to 3D cell cultures. The detection system has a potential as a single package to investigate 3D cultures in a microfluidic system.

© 2010 Optical Society of America

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J. H. Sung, J. Choi, D. Kim, and M. L. Shuler, Biotechnol. Bioeng. 104, 516 (2009).
[CrossRef] [PubMed]

S. Toetsch, P. Olwell, A. Prina-Mello, and Y. Volkov,Integr. Biol. 1, 170 (2009)
[CrossRef]

2008

N. B. E. Sawyer, L. K. Worrall, J. A. Crowe, S. L. Waters, K. M. Shakesheff, F. R. A. J. Rose, and S. P. Morgan, Biotechnol. Bioeng. 100, 159 (2008).
[CrossRef]

A. P. Wong, R. Perez-Castillejos, J. C. Love, and G. M. Whitesides, Biomaterials 29, 1853 (2008).
[CrossRef] [PubMed]

2007

F. Pampaloni, E. G. Reynaud, and E. H. K. Stelzer, Nat. Rev. Mol. Cell Biol. 8, 839 (2007).
[CrossRef] [PubMed]

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[CrossRef] [PubMed]

T. Oh, J. H. Sung, D. A. Tatosian, M. L. Shuler, and D. Kim, Cytometry, Part A 71A, 857 (2007).
[CrossRef]

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2005

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A. Abbott, Nature 424, 870 (2003).
[CrossRef] [PubMed]

Abbott, A.

A. Abbott, Nature 424, 870 (2003).
[CrossRef] [PubMed]

Bissell, M. J.

G. Y. Lee, P. A. Kenny, E. H. Lee, and M. J. Bissell, Nat. Methods 4, 359 (2007).
[CrossRef] [PubMed]

Boppart, S. A.

Choi, J.

J. H. Sung, J. Choi, D. Kim, and M. L. Shuler, Biotechnol. Bioeng. 104, 516 (2009).
[CrossRef] [PubMed]

Crowe, J. A.

N. B. E. Sawyer, L. K. Worrall, J. A. Crowe, S. L. Waters, K. M. Shakesheff, F. R. A. J. Rose, and S. P. Morgan, Biotechnol. Bioeng. 100, 159 (2008).
[CrossRef]

Desai, T. A.

Kenny, P. A.

G. Y. Lee, P. A. Kenny, E. H. Lee, and M. J. Bissell, Nat. Methods 4, 359 (2007).
[CrossRef] [PubMed]

Kim, D.

J. H. Sung, J. Choi, D. Kim, and M. L. Shuler, Biotechnol. Bioeng. 104, 516 (2009).
[CrossRef] [PubMed]

T. Oh, J. H. Sung, D. A. Tatosian, M. L. Shuler, and D. Kim, Cytometry, Part A 71A, 857 (2007).
[CrossRef]

D. Tatosian, M. L. Shuler, and D. Kim, Opt. Lett. 30, 1689 (2005).
[CrossRef] [PubMed]

Kwon, S.

Lee, E. H.

G. Y. Lee, P. A. Kenny, E. H. Lee, and M. J. Bissell, Nat. Methods 4, 359 (2007).
[CrossRef] [PubMed]

Lee, G. Y.

G. Y. Lee, P. A. Kenny, E. H. Lee, and M. J. Bissell, Nat. Methods 4, 359 (2007).
[CrossRef] [PubMed]

Lee, L. P.

Love, J. C.

A. P. Wong, R. Perez-Castillejos, J. C. Love, and G. M. Whitesides, Biomaterials 29, 1853 (2008).
[CrossRef] [PubMed]

Morgan, S. P.

N. B. E. Sawyer, L. K. Worrall, J. A. Crowe, S. L. Waters, K. M. Shakesheff, F. R. A. J. Rose, and S. P. Morgan, Biotechnol. Bioeng. 100, 159 (2008).
[CrossRef]

Norman, J. J.

Oh, T.

T. Oh, J. H. Sung, D. A. Tatosian, M. L. Shuler, and D. Kim, Cytometry, Part A 71A, 857 (2007).
[CrossRef]

Oldenburg, A. L.

Olwell, P.

S. Toetsch, P. Olwell, A. Prina-Mello, and Y. Volkov,Integr. Biol. 1, 170 (2009)
[CrossRef]

Pampaloni, F.

F. Pampaloni, E. G. Reynaud, and E. H. K. Stelzer, Nat. Rev. Mol. Cell Biol. 8, 839 (2007).
[CrossRef] [PubMed]

Perez-Castillejos, R.

A. P. Wong, R. Perez-Castillejos, J. C. Love, and G. M. Whitesides, Biomaterials 29, 1853 (2008).
[CrossRef] [PubMed]

Prina-Mello, A.

S. Toetsch, P. Olwell, A. Prina-Mello, and Y. Volkov,Integr. Biol. 1, 170 (2009)
[CrossRef]

Reynaud, E. G.

F. Pampaloni, E. G. Reynaud, and E. H. K. Stelzer, Nat. Rev. Mol. Cell Biol. 8, 839 (2007).
[CrossRef] [PubMed]

Rose, F. R. A. J.

N. B. E. Sawyer, L. K. Worrall, J. A. Crowe, S. L. Waters, K. M. Shakesheff, F. R. A. J. Rose, and S. P. Morgan, Biotechnol. Bioeng. 100, 159 (2008).
[CrossRef]

Sawyer, N. B. E.

N. B. E. Sawyer, L. K. Worrall, J. A. Crowe, S. L. Waters, K. M. Shakesheff, F. R. A. J. Rose, and S. P. Morgan, Biotechnol. Bioeng. 100, 159 (2008).
[CrossRef]

Shakesheff, K. M.

N. B. E. Sawyer, L. K. Worrall, J. A. Crowe, S. L. Waters, K. M. Shakesheff, F. R. A. J. Rose, and S. P. Morgan, Biotechnol. Bioeng. 100, 159 (2008).
[CrossRef]

Shuler, M. L.

J. H. Sung, J. Choi, D. Kim, and M. L. Shuler, Biotechnol. Bioeng. 104, 516 (2009).
[CrossRef] [PubMed]

T. Oh, J. H. Sung, D. A. Tatosian, M. L. Shuler, and D. Kim, Cytometry, Part A 71A, 857 (2007).
[CrossRef]

D. Tatosian, M. L. Shuler, and D. Kim, Opt. Lett. 30, 1689 (2005).
[CrossRef] [PubMed]

Stelzer, E. H. K.

F. Pampaloni, E. G. Reynaud, and E. H. K. Stelzer, Nat. Rev. Mol. Cell Biol. 8, 839 (2007).
[CrossRef] [PubMed]

Sung, J. H.

J. H. Sung, J. Choi, D. Kim, and M. L. Shuler, Biotechnol. Bioeng. 104, 516 (2009).
[CrossRef] [PubMed]

T. Oh, J. H. Sung, D. A. Tatosian, M. L. Shuler, and D. Kim, Cytometry, Part A 71A, 857 (2007).
[CrossRef]

Tan, W.

Tatosian, D.

Tatosian, D. A.

T. Oh, J. H. Sung, D. A. Tatosian, M. L. Shuler, and D. Kim, Cytometry, Part A 71A, 857 (2007).
[CrossRef]

Toetsch, S.

S. Toetsch, P. Olwell, A. Prina-Mello, and Y. Volkov,Integr. Biol. 1, 170 (2009)
[CrossRef]

Volkov, Y.

S. Toetsch, P. Olwell, A. Prina-Mello, and Y. Volkov,Integr. Biol. 1, 170 (2009)
[CrossRef]

Waters, S. L.

N. B. E. Sawyer, L. K. Worrall, J. A. Crowe, S. L. Waters, K. M. Shakesheff, F. R. A. J. Rose, and S. P. Morgan, Biotechnol. Bioeng. 100, 159 (2008).
[CrossRef]

Whitesides, G. M.

A. P. Wong, R. Perez-Castillejos, J. C. Love, and G. M. Whitesides, Biomaterials 29, 1853 (2008).
[CrossRef] [PubMed]

Wong, A. P.

A. P. Wong, R. Perez-Castillejos, J. C. Love, and G. M. Whitesides, Biomaterials 29, 1853 (2008).
[CrossRef] [PubMed]

Worrall, L. K.

N. B. E. Sawyer, L. K. Worrall, J. A. Crowe, S. L. Waters, K. M. Shakesheff, F. R. A. J. Rose, and S. P. Morgan, Biotechnol. Bioeng. 100, 159 (2008).
[CrossRef]

Biomaterials

A. P. Wong, R. Perez-Castillejos, J. C. Love, and G. M. Whitesides, Biomaterials 29, 1853 (2008).
[CrossRef] [PubMed]

Biotechnol. Bioeng.

J. H. Sung, J. Choi, D. Kim, and M. L. Shuler, Biotechnol. Bioeng. 104, 516 (2009).
[CrossRef] [PubMed]

N. B. E. Sawyer, L. K. Worrall, J. A. Crowe, S. L. Waters, K. M. Shakesheff, F. R. A. J. Rose, and S. P. Morgan, Biotechnol. Bioeng. 100, 159 (2008).
[CrossRef]

Cytometry, Part A

T. Oh, J. H. Sung, D. A. Tatosian, M. L. Shuler, and D. Kim, Cytometry, Part A 71A, 857 (2007).
[CrossRef]

Integr. Biol.

S. Toetsch, P. Olwell, A. Prina-Mello, and Y. Volkov,Integr. Biol. 1, 170 (2009)
[CrossRef]

Nat. Methods

G. Y. Lee, P. A. Kenny, E. H. Lee, and M. J. Bissell, Nat. Methods 4, 359 (2007).
[CrossRef] [PubMed]

Nat. Rev. Mol. Cell Biol.

F. Pampaloni, E. G. Reynaud, and E. H. K. Stelzer, Nat. Rev. Mol. Cell Biol. 8, 839 (2007).
[CrossRef] [PubMed]

Nature

A. Abbott, Nature 424, 870 (2003).
[CrossRef] [PubMed]

Opt. Express

Opt. Lett.

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Figures (4)

Fig. 1
Fig. 1

(a) Optical setup for the used 3D microfluidic assay (OB, objective; P, pinhole; F, filter; and DM, dichroic mirror). Also shown are the schematics of the 3D cell culture assays for studies of (b) cell staining, (c) cell cytotoxicity, and (d) cell invasion (not scaled).

Fig. 2
Fig. 2

Fluorescent intensity measured by the 3D-ISFODS for cell-stained solution (celltracker orange) applied on the 3D alginate-cell matrix at t = 0   min . Solid curves represent calculated trends based on a transport model with diffusivity D = 2.4 × 10 10 m 2 / s .

Fig. 3
Fig. 3

Fluorescent intensity measured with cell cytotoxicity solution applied at t = 20   min . Diffusivity was assumed as D = 2.4 × 10 10 m 2 / s .

Fig. 4
Fig. 4

Fluorescent intensity measured for cell-invasion assay cultured in Matrigel and alginate matrix.

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