Abstract

We demonstrate an inverse light-scattering analysis procedure based on using the T-matrix method as a light-scattering model. We measure light scattered by in vitro cell monolayers using angle-resolved low-coherence interferometry (a/LCI) and compare the data to predictions of the T-matrix theory. The comparison yields measurements of the equal volume diameter and aspect ratio of the spheroid cell nuclei with accuracy comparable to quantitative image analysis of fixed and stained samples. These improvements represent a significant upgrade for the a/LCI technique, expanding both the range of tissue in which it is applicable and potentially increasing its value as a diagnostic tool.

© 2008 Optical Society of America

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  1. A. Wax, J. W. Pyhtila, R. N. Graf, R. Nines, C. W. Boone, R. R. Dasari, M. S. Feld, V. E. Steele, and G. D. Stoner, J. Biomed. Opt. 10, 051604 (2005).
    [CrossRef] [PubMed]
  2. K. J. Chalut, L. A. Kresty, J. W. Pyhtila, R. Nines, M. Baird, V. E. Steele, and A. Wax, Cancer Epidemiol. Biomarkers Prev. 16, 223 (2007).
    [CrossRef] [PubMed]
  3. J. W. Pyhtila, R. N. Graf, and A. Wax, Opt. Express 11, 3473 (2003).
    [CrossRef] [PubMed]
  4. K. J. Chalut, M. G. Giacomelli, and A. Wax, J. Opt. Soc. Am. A 25, 1866 (2008).
  5. K. Chalut, S. Chen, J. Finan, M. Giacomelli, F. Guilak, K. Leong, and A. Wax, Biophys. J. 107, 124107 (2008).
  6. M. I. Mishchenko, L. D. Travis, and D. W. Mackowski, J. Quant. Spectrosc. Radiat. Transf. 55, 535 (1996).
    [CrossRef]
  7. M. I. Mishchenko, L. D. Travis, and J. W. Hovenier, Light Scattering by Nonspherical Particles: Theory, Measurements and Applications (Academic, 2000).
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    [CrossRef]
  9. J. D. Keener, K. J. Chalut, J. W. Pyhtila, and A. Wax, Opt. Lett. 32, 1326 (2007).
    [CrossRef] [PubMed]
  10. J. R. Mourant, T. Aida, L. Coburn, and J. Ramachandran, Proc. SPIE 5326, 54 (2004).
    [CrossRef]
  11. D. D. Duncan, and M. E. Thomas, Appl. Opt. 46, 6185 (2007).
    [CrossRef] [PubMed]
  12. D. Zink, A. H. Fischer, and J. A. Nickerson, Nat. Rev. Cancer 8, 677 (2004).
    [CrossRef]

2008 (2)

K. J. Chalut, M. G. Giacomelli, and A. Wax, J. Opt. Soc. Am. A 25, 1866 (2008).

K. Chalut, S. Chen, J. Finan, M. Giacomelli, F. Guilak, K. Leong, and A. Wax, Biophys. J. 107, 124107 (2008).

2007 (3)

K. J. Chalut, L. A. Kresty, J. W. Pyhtila, R. Nines, M. Baird, V. E. Steele, and A. Wax, Cancer Epidemiol. Biomarkers Prev. 16, 223 (2007).
[CrossRef] [PubMed]

J. D. Keener, K. J. Chalut, J. W. Pyhtila, and A. Wax, Opt. Lett. 32, 1326 (2007).
[CrossRef] [PubMed]

D. D. Duncan, and M. E. Thomas, Appl. Opt. 46, 6185 (2007).
[CrossRef] [PubMed]

2005 (1)

A. Wax, J. W. Pyhtila, R. N. Graf, R. Nines, C. W. Boone, R. R. Dasari, M. S. Feld, V. E. Steele, and G. D. Stoner, J. Biomed. Opt. 10, 051604 (2005).
[CrossRef] [PubMed]

2004 (2)

J. R. Mourant, T. Aida, L. Coburn, and J. Ramachandran, Proc. SPIE 5326, 54 (2004).
[CrossRef]

D. Zink, A. H. Fischer, and J. A. Nickerson, Nat. Rev. Cancer 8, 677 (2004).
[CrossRef]

2003 (1)

1998 (1)

1996 (1)

M. I. Mishchenko, L. D. Travis, and D. W. Mackowski, J. Quant. Spectrosc. Radiat. Transf. 55, 535 (1996).
[CrossRef]

Aida, T.

J. R. Mourant, T. Aida, L. Coburn, and J. Ramachandran, Proc. SPIE 5326, 54 (2004).
[CrossRef]

Baird, M.

K. J. Chalut, L. A. Kresty, J. W. Pyhtila, R. Nines, M. Baird, V. E. Steele, and A. Wax, Cancer Epidemiol. Biomarkers Prev. 16, 223 (2007).
[CrossRef] [PubMed]

Boone, C. W.

A. Wax, J. W. Pyhtila, R. N. Graf, R. Nines, C. W. Boone, R. R. Dasari, M. S. Feld, V. E. Steele, and G. D. Stoner, J. Biomed. Opt. 10, 051604 (2005).
[CrossRef] [PubMed]

Chalut, K.

K. Chalut, S. Chen, J. Finan, M. Giacomelli, F. Guilak, K. Leong, and A. Wax, Biophys. J. 107, 124107 (2008).

Chalut, K. J.

K. J. Chalut, M. G. Giacomelli, and A. Wax, J. Opt. Soc. Am. A 25, 1866 (2008).

J. D. Keener, K. J. Chalut, J. W. Pyhtila, and A. Wax, Opt. Lett. 32, 1326 (2007).
[CrossRef] [PubMed]

K. J. Chalut, L. A. Kresty, J. W. Pyhtila, R. Nines, M. Baird, V. E. Steele, and A. Wax, Cancer Epidemiol. Biomarkers Prev. 16, 223 (2007).
[CrossRef] [PubMed]

Chen, S.

K. Chalut, S. Chen, J. Finan, M. Giacomelli, F. Guilak, K. Leong, and A. Wax, Biophys. J. 107, 124107 (2008).

Coburn, L.

J. R. Mourant, T. Aida, L. Coburn, and J. Ramachandran, Proc. SPIE 5326, 54 (2004).
[CrossRef]

Dasari, R. R.

A. Wax, J. W. Pyhtila, R. N. Graf, R. Nines, C. W. Boone, R. R. Dasari, M. S. Feld, V. E. Steele, and G. D. Stoner, J. Biomed. Opt. 10, 051604 (2005).
[CrossRef] [PubMed]

Duncan, D. D.

Feld, M. S.

A. Wax, J. W. Pyhtila, R. N. Graf, R. Nines, C. W. Boone, R. R. Dasari, M. S. Feld, V. E. Steele, and G. D. Stoner, J. Biomed. Opt. 10, 051604 (2005).
[CrossRef] [PubMed]

Finan, J.

K. Chalut, S. Chen, J. Finan, M. Giacomelli, F. Guilak, K. Leong, and A. Wax, Biophys. J. 107, 124107 (2008).

Fischer, A. H.

D. Zink, A. H. Fischer, and J. A. Nickerson, Nat. Rev. Cancer 8, 677 (2004).
[CrossRef]

Giacomelli, M.

K. Chalut, S. Chen, J. Finan, M. Giacomelli, F. Guilak, K. Leong, and A. Wax, Biophys. J. 107, 124107 (2008).

Giacomelli, M. G.

Graf, R. N.

A. Wax, J. W. Pyhtila, R. N. Graf, R. Nines, C. W. Boone, R. R. Dasari, M. S. Feld, V. E. Steele, and G. D. Stoner, J. Biomed. Opt. 10, 051604 (2005).
[CrossRef] [PubMed]

J. W. Pyhtila, R. N. Graf, and A. Wax, Opt. Express 11, 3473 (2003).
[CrossRef] [PubMed]

Guilak, F.

K. Chalut, S. Chen, J. Finan, M. Giacomelli, F. Guilak, K. Leong, and A. Wax, Biophys. J. 107, 124107 (2008).

Hovenier, J. W.

M. I. Mishchenko, L. D. Travis, and J. W. Hovenier, Light Scattering by Nonspherical Particles: Theory, Measurements and Applications (Academic, 2000).

Keener, J. D.

Kresty, L. A.

K. J. Chalut, L. A. Kresty, J. W. Pyhtila, R. Nines, M. Baird, V. E. Steele, and A. Wax, Cancer Epidemiol. Biomarkers Prev. 16, 223 (2007).
[CrossRef] [PubMed]

Leong, K.

K. Chalut, S. Chen, J. Finan, M. Giacomelli, F. Guilak, K. Leong, and A. Wax, Biophys. J. 107, 124107 (2008).

Mackowski, D. W.

M. I. Mishchenko, L. D. Travis, and D. W. Mackowski, J. Quant. Spectrosc. Radiat. Transf. 55, 535 (1996).
[CrossRef]

Mishchenko, M. I.

M. I. Mishchenko, L. D. Travis, and D. W. Mackowski, J. Quant. Spectrosc. Radiat. Transf. 55, 535 (1996).
[CrossRef]

M. I. Mishchenko, L. D. Travis, and J. W. Hovenier, Light Scattering by Nonspherical Particles: Theory, Measurements and Applications (Academic, 2000).

Mourant, J. R.

J. R. Mourant, T. Aida, L. Coburn, and J. Ramachandran, Proc. SPIE 5326, 54 (2004).
[CrossRef]

Nickerson, J. A.

D. Zink, A. H. Fischer, and J. A. Nickerson, Nat. Rev. Cancer 8, 677 (2004).
[CrossRef]

Nilsson, A. M. K.

Nines, R.

K. J. Chalut, L. A. Kresty, J. W. Pyhtila, R. Nines, M. Baird, V. E. Steele, and A. Wax, Cancer Epidemiol. Biomarkers Prev. 16, 223 (2007).
[CrossRef] [PubMed]

A. Wax, J. W. Pyhtila, R. N. Graf, R. Nines, C. W. Boone, R. R. Dasari, M. S. Feld, V. E. Steele, and G. D. Stoner, J. Biomed. Opt. 10, 051604 (2005).
[CrossRef] [PubMed]

Pyhtila, J. W.

K. J. Chalut, L. A. Kresty, J. W. Pyhtila, R. Nines, M. Baird, V. E. Steele, and A. Wax, Cancer Epidemiol. Biomarkers Prev. 16, 223 (2007).
[CrossRef] [PubMed]

J. D. Keener, K. J. Chalut, J. W. Pyhtila, and A. Wax, Opt. Lett. 32, 1326 (2007).
[CrossRef] [PubMed]

A. Wax, J. W. Pyhtila, R. N. Graf, R. Nines, C. W. Boone, R. R. Dasari, M. S. Feld, V. E. Steele, and G. D. Stoner, J. Biomed. Opt. 10, 051604 (2005).
[CrossRef] [PubMed]

J. W. Pyhtila, R. N. Graf, and A. Wax, Opt. Express 11, 3473 (2003).
[CrossRef] [PubMed]

Ramachandran, J.

J. R. Mourant, T. Aida, L. Coburn, and J. Ramachandran, Proc. SPIE 5326, 54 (2004).
[CrossRef]

Steele, V. E.

K. J. Chalut, L. A. Kresty, J. W. Pyhtila, R. Nines, M. Baird, V. E. Steele, and A. Wax, Cancer Epidemiol. Biomarkers Prev. 16, 223 (2007).
[CrossRef] [PubMed]

A. Wax, J. W. Pyhtila, R. N. Graf, R. Nines, C. W. Boone, R. R. Dasari, M. S. Feld, V. E. Steele, and G. D. Stoner, J. Biomed. Opt. 10, 051604 (2005).
[CrossRef] [PubMed]

Stoner, G. D.

A. Wax, J. W. Pyhtila, R. N. Graf, R. Nines, C. W. Boone, R. R. Dasari, M. S. Feld, V. E. Steele, and G. D. Stoner, J. Biomed. Opt. 10, 051604 (2005).
[CrossRef] [PubMed]

Thomas, M. E.

Travis, L. D.

M. I. Mishchenko, L. D. Travis, and D. W. Mackowski, J. Quant. Spectrosc. Radiat. Transf. 55, 535 (1996).
[CrossRef]

M. I. Mishchenko, L. D. Travis, and J. W. Hovenier, Light Scattering by Nonspherical Particles: Theory, Measurements and Applications (Academic, 2000).

Wax, A.

K. J. Chalut, M. G. Giacomelli, and A. Wax, J. Opt. Soc. Am. A 25, 1866 (2008).

K. Chalut, S. Chen, J. Finan, M. Giacomelli, F. Guilak, K. Leong, and A. Wax, Biophys. J. 107, 124107 (2008).

J. D. Keener, K. J. Chalut, J. W. Pyhtila, and A. Wax, Opt. Lett. 32, 1326 (2007).
[CrossRef] [PubMed]

K. J. Chalut, L. A. Kresty, J. W. Pyhtila, R. Nines, M. Baird, V. E. Steele, and A. Wax, Cancer Epidemiol. Biomarkers Prev. 16, 223 (2007).
[CrossRef] [PubMed]

A. Wax, J. W. Pyhtila, R. N. Graf, R. Nines, C. W. Boone, R. R. Dasari, M. S. Feld, V. E. Steele, and G. D. Stoner, J. Biomed. Opt. 10, 051604 (2005).
[CrossRef] [PubMed]

J. W. Pyhtila, R. N. Graf, and A. Wax, Opt. Express 11, 3473 (2003).
[CrossRef] [PubMed]

Zink, D.

D. Zink, A. H. Fischer, and J. A. Nickerson, Nat. Rev. Cancer 8, 677 (2004).
[CrossRef]

Appl. Opt. (2)

Biophys. J. (1)

K. Chalut, S. Chen, J. Finan, M. Giacomelli, F. Guilak, K. Leong, and A. Wax, Biophys. J. 107, 124107 (2008).

Cancer Epidemiol. Biomarkers Prev. (1)

K. J. Chalut, L. A. Kresty, J. W. Pyhtila, R. Nines, M. Baird, V. E. Steele, and A. Wax, Cancer Epidemiol. Biomarkers Prev. 16, 223 (2007).
[CrossRef] [PubMed]

J. Biomed. Opt. (1)

A. Wax, J. W. Pyhtila, R. N. Graf, R. Nines, C. W. Boone, R. R. Dasari, M. S. Feld, V. E. Steele, and G. D. Stoner, J. Biomed. Opt. 10, 051604 (2005).
[CrossRef] [PubMed]

J. Opt. Soc. Am. A (1)

J. Quant. Spectrosc. Radiat. Transf. (1)

M. I. Mishchenko, L. D. Travis, and D. W. Mackowski, J. Quant. Spectrosc. Radiat. Transf. 55, 535 (1996).
[CrossRef]

Nat. Rev. Cancer (1)

D. Zink, A. H. Fischer, and J. A. Nickerson, Nat. Rev. Cancer 8, 677 (2004).
[CrossRef]

Opt. Express (1)

Opt. Lett. (1)

Proc. SPIE (1)

J. R. Mourant, T. Aida, L. Coburn, and J. Ramachandran, Proc. SPIE 5326, 54 (2004).
[CrossRef]

Other (1)

M. I. Mishchenko, L. D. Travis, and J. W. Hovenier, Light Scattering by Nonspherical Particles: Theory, Measurements and Applications (Academic, 2000).

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Figures (3)

Fig. 1
Fig. 1

(A) a/LCI measurement showing depth-resolved angular scattering for a MCF7 cell monolayer with angles given relative to the backscatter angle. (B) Unprocessed angular scattering profile extracted by summing over depth in (A). (C) Processed scattering profile compared to best fit T-matrix model after detrending and low-pass filtering to isolate nuclear scattering.

Fig. 2
Fig. 2

Fluorescence microscopy image of MCF7 nuclei using DAPI. Nuclei are approximately spheroidal and randomly aligned.

Fig. 3
Fig. 3

Measurements of equal volume diameter and spheriodial aspect ratio by QIA using DAPI stain, and a/LCI using live cells. Error bars are standard error at 95% confidence.

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