Abstract

We present a Mie theory-based analysis of angularly resolved light scattering from intact cells that involves the product of a particle size distribution ρ and the scattering cross section σ. The analysis shows that the ability to size intracellular organelles depends strongly upon the assumed functional form of the particle size distribution; however, the product σρ extracts the dominant light scatterers regardless of this choice. We examine goniometer measurements from our own and another laboratory and find that this analysis reconciles seemingly conflicting results and confirms that mitochondrial-sized organelles dominate scattering from intact cells in the angular range 5°–90°.

© 2005 Optical Society of America

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  1. L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
    [CrossRef]
  2. A. Wax, C. Yang, V. Backman, K. Badizadegan, C. W. Boone, R. R. Dasari, and M. S. Feld, Biophys. J. 82, 2256 (2002).
    [CrossRef] [PubMed]
  3. J. R. Mourant, T. M. Johnson, S. Carpenter, A. Guerra, T. Aida, and J. P. Freyer, J. Biomed. Opt. 7, 378 (2002).
    [CrossRef] [PubMed]
  4. J. D. Wilson, C. E. Bigelow, D. J. Calkins, and T. H. Foster, Biophys. J. 88, 2929 (2005).
    [CrossRef] [PubMed]
  5. C. F. Bohren and D. R. Huffman, Absorption and Scattering of Light from Small Particles (Wiley, 1998).
    [CrossRef]
  6. R. Drezek, A. Dunn, and R. Richards-Kortum, Appl. Opt. 38, 3651 (1999).
    [CrossRef]
  7. G. E. Palade, J. Histochem. Cytochem. 1, 188 (1953).
    [CrossRef] [PubMed]

2005

J. D. Wilson, C. E. Bigelow, D. J. Calkins, and T. H. Foster, Biophys. J. 88, 2929 (2005).
[CrossRef] [PubMed]

2002

A. Wax, C. Yang, V. Backman, K. Badizadegan, C. W. Boone, R. R. Dasari, and M. S. Feld, Biophys. J. 82, 2256 (2002).
[CrossRef] [PubMed]

J. R. Mourant, T. M. Johnson, S. Carpenter, A. Guerra, T. Aida, and J. P. Freyer, J. Biomed. Opt. 7, 378 (2002).
[CrossRef] [PubMed]

1999

1998

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

1953

G. E. Palade, J. Histochem. Cytochem. 1, 188 (1953).
[CrossRef] [PubMed]

Aida, T.

J. R. Mourant, T. M. Johnson, S. Carpenter, A. Guerra, T. Aida, and J. P. Freyer, J. Biomed. Opt. 7, 378 (2002).
[CrossRef] [PubMed]

Backman, V.

A. Wax, C. Yang, V. Backman, K. Badizadegan, C. W. Boone, R. R. Dasari, and M. S. Feld, Biophys. J. 82, 2256 (2002).
[CrossRef] [PubMed]

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Badizadegan, K.

A. Wax, C. Yang, V. Backman, K. Badizadegan, C. W. Boone, R. R. Dasari, and M. S. Feld, Biophys. J. 82, 2256 (2002).
[CrossRef] [PubMed]

Bigelow, C. E.

J. D. Wilson, C. E. Bigelow, D. J. Calkins, and T. H. Foster, Biophys. J. 88, 2929 (2005).
[CrossRef] [PubMed]

Bohren, C. F.

C. F. Bohren and D. R. Huffman, Absorption and Scattering of Light from Small Particles (Wiley, 1998).
[CrossRef]

Boone, C. W.

A. Wax, C. Yang, V. Backman, K. Badizadegan, C. W. Boone, R. R. Dasari, and M. S. Feld, Biophys. J. 82, 2256 (2002).
[CrossRef] [PubMed]

Calkins, D. J.

J. D. Wilson, C. E. Bigelow, D. J. Calkins, and T. H. Foster, Biophys. J. 88, 2929 (2005).
[CrossRef] [PubMed]

Carpenter, S.

J. R. Mourant, T. M. Johnson, S. Carpenter, A. Guerra, T. Aida, and J. P. Freyer, J. Biomed. Opt. 7, 378 (2002).
[CrossRef] [PubMed]

Crawford, J. M.

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Dasari, R. R.

A. Wax, C. Yang, V. Backman, K. Badizadegan, C. W. Boone, R. R. Dasari, and M. S. Feld, Biophys. J. 82, 2256 (2002).
[CrossRef] [PubMed]

Drezek, R.

Dunn, A.

Feld, M. S.

A. Wax, C. Yang, V. Backman, K. Badizadegan, C. W. Boone, R. R. Dasari, and M. S. Feld, Biophys. J. 82, 2256 (2002).
[CrossRef] [PubMed]

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Foster, T. H.

J. D. Wilson, C. E. Bigelow, D. J. Calkins, and T. H. Foster, Biophys. J. 88, 2929 (2005).
[CrossRef] [PubMed]

Freyer, J. P.

J. R. Mourant, T. M. Johnson, S. Carpenter, A. Guerra, T. Aida, and J. P. Freyer, J. Biomed. Opt. 7, 378 (2002).
[CrossRef] [PubMed]

Guerra, A.

J. R. Mourant, T. M. Johnson, S. Carpenter, A. Guerra, T. Aida, and J. P. Freyer, J. Biomed. Opt. 7, 378 (2002).
[CrossRef] [PubMed]

Hamano, T.

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Huffman, D. R.

C. F. Bohren and D. R. Huffman, Absorption and Scattering of Light from Small Particles (Wiley, 1998).
[CrossRef]

Itzkan, I.

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Johnson, T. M.

J. R. Mourant, T. M. Johnson, S. Carpenter, A. Guerra, T. Aida, and J. P. Freyer, J. Biomed. Opt. 7, 378 (2002).
[CrossRef] [PubMed]

Lima, C.

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Manoharan, R.

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Mourant, J. R.

J. R. Mourant, T. M. Johnson, S. Carpenter, A. Guerra, T. Aida, and J. P. Freyer, J. Biomed. Opt. 7, 378 (2002).
[CrossRef] [PubMed]

Nusrat, A.

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Palade, G. E.

G. E. Palade, J. Histochem. Cytochem. 1, 188 (1953).
[CrossRef] [PubMed]

Perelman, L. T.

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Richards-Kortum, R.

Seiler, M.

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Shields, S.

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Van Dam, J.

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Wallace, M.

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Wax, A.

A. Wax, C. Yang, V. Backman, K. Badizadegan, C. W. Boone, R. R. Dasari, and M. S. Feld, Biophys. J. 82, 2256 (2002).
[CrossRef] [PubMed]

Wilson, J. D.

J. D. Wilson, C. E. Bigelow, D. J. Calkins, and T. H. Foster, Biophys. J. 88, 2929 (2005).
[CrossRef] [PubMed]

Yang, C.

A. Wax, C. Yang, V. Backman, K. Badizadegan, C. W. Boone, R. R. Dasari, and M. S. Feld, Biophys. J. 82, 2256 (2002).
[CrossRef] [PubMed]

Zonios, G.

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Appl. Opt.

Biophys. J.

A. Wax, C. Yang, V. Backman, K. Badizadegan, C. W. Boone, R. R. Dasari, and M. S. Feld, Biophys. J. 82, 2256 (2002).
[CrossRef] [PubMed]

J. D. Wilson, C. E. Bigelow, D. J. Calkins, and T. H. Foster, Biophys. J. 88, 2929 (2005).
[CrossRef] [PubMed]

J. Biomed. Opt.

J. R. Mourant, T. M. Johnson, S. Carpenter, A. Guerra, T. Aida, and J. P. Freyer, J. Biomed. Opt. 7, 378 (2002).
[CrossRef] [PubMed]

J. Histochem. Cytochem.

G. E. Palade, J. Histochem. Cytochem. 1, 188 (1953).
[CrossRef] [PubMed]

Phys. Rev. Lett.

L. T. Perelman, V. Backman, M. Wallace, G. Zonios, R. Manoharan, A. Nusrat, S. Shields, M. Seiler, C. Lima, T. Hamano, I. Itzkan, J. Van Dam, J. M. Crawford, and M. S. Feld, Phys. Rev. Lett. 80, 627 (1998).
[CrossRef]

Other

C. F. Bohren and D. R. Huffman, Absorption and Scattering of Light from Small Particles (Wiley, 1998).
[CrossRef]

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Figures (4)

Fig. 1
Fig. 1

Relationship between the scattering cross-section-weighted size distribution, σ ρ (solid curve), and the particle size distribution, ρ ( r ) (dashed curve). The log normal particle size distribution shown here has a mean diameter and standard deviation of 1.09 and 0.6 μ m , respectively. The scattering cross section, σ ( r ) (dotted curve), as a function of particle diameter is plotted for spheres of refractive index 1.40 in a surrounding medium of index 1.38 and a vacuum wavelength of 633 nm.

Fig. 2
Fig. 2

Angularly resolved light scattering data (open circles) from intact EMT6 cells with bimodal fits (solid curves) assuming A, Gaussian, B, exponential, and C, log normal distributions. Data were obtained at 31 angular positions from 5° to 90°. The global minimum in χ 2 was obtained with the log normal fit. Mean scatterer diameters, standard deviations, and relative numbers of scatterer populations are summarized in Table 1 for the three assumed particle size distributions.

Fig. 3
Fig. 3

Relative total light scattered ( σ ρ ) versus particle diameter for the three distributions used to fit the data of Fig. 2 and a fourth curve resulting from a fit using a log normal functional form for σ ρ with a trimodal distribution. The curves are normalized to 1 at their peak value.

Fig. 4
Fig. 4

A, Bimodal size distribution of Fig. 6 from Mourant et al.,[3] which is a particle size distribution returned from a Mie theory fit to angularly resolved scattering data from AT3.1 cells. Note that this is plotted on a log scale, and the number of large scatterers is much smaller than the number of small scatterers. B, Distribution shown in A weighted by the scattering cross section (solid curve) plotted along with the total light scattered returned from the log normal fits to scattering from EMT6 cells (dashed curve) depicted in Fig. 3.

Tables (1)

Tables Icon

Table 1 Parameters Returned from Mie Theory Fits to Angularly Resolved Scattering Data Using Three Functional Forms of Particle Size Distributions a

Equations (2)

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P total ( θ ) = ( j = 1 N σ j P j ( θ ) j = 1 N σ j ) .
P total ( θ ) = σ ( r ) ρ ( r ) P ( r , θ ) d r σ ( r ) ρ ( r ) d r .

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