Abstract

Digital holographic microscopy (DHM) allows optical-path-difference (OPD) measurements with nanometric accuracy. OPD induced by transparent cells depends on both the refractive index (RI) of cells and their morphology. This Letter presents a dual-wavelength DHM that allows us to separately measure both the RI and the cellular thickness by exploiting an enhanced dispersion of the perfusion medium achieved by the utilization of an extracellular dye. The two wavelengths are chosen in the vicinity of the absorption peak of the dye, where the absorption is accompanied by a significant variation of the RI as a function of the wavelength.

© 2008 Optical Society of America

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2007 (1)

2006 (3)

2005 (5)

C. L. Curl, C. J. Bellair, T. Harris, B. E. Allman, P. J. Harris, A. G. Stewart, A. Roberts, K. A. Nugent, and L. M. Delbridge, Cytometry 65, 88 (2005).
[CrossRef] [PubMed]

V. Lucarini, J. J. Saarinen, K.-E. Peiponen, and E. M. Vartiainen, Kramers-Kronig Relations in Optical Materials Research (Springer, 2005).

P. Marquet, B. Rappaz, P. J. Magistretti, E. Cuche, Y. Emery, T. Colomb, and C. Depeursinge, Opt. Lett. 30, 468 (2005).
[CrossRef] [PubMed]

C. Mann, L. Yu, C.-M. Lo, and M. Kim, Opt. Express 13, 8693 (2005).
[CrossRef] [PubMed]

B. Rappaz, P. Marquet, E. Cuche, Y. Emery, C. Depeursinge, and P. J. Magistretti, Opt. Express 13, 9361 (2005).
[CrossRef] [PubMed]

1969 (1)

R. Barer, J. Histochem. Cytochem. 17, 423 (1969).
[CrossRef] [PubMed]

1967 (1)

K. A. Chaubal, Z. Lodin, and J. Pilny, Acta Histochem. 26, 131 (1967).
[PubMed]

Allman, B. E.

C. L. Curl, C. J. Bellair, T. Harris, B. E. Allman, P. J. Harris, A. G. Stewart, A. Roberts, K. A. Nugent, and L. M. Delbridge, Cytometry 65, 88 (2005).
[CrossRef] [PubMed]

Aspert, N.

Badizadegan, K.

Barer, R.

R. Barer, J. Histochem. Cytochem. 17, 423 (1969).
[CrossRef] [PubMed]

Bellair, C. J.

C. L. Curl, C. J. Bellair, T. Harris, B. E. Allman, P. J. Harris, A. G. Stewart, A. Roberts, K. A. Nugent, and L. M. Delbridge, Cytometry 65, 88 (2005).
[CrossRef] [PubMed]

Bourquin, S.

Bredebusch, I.

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schafer, W. Domschke, and G. von Bally, J. Biomed. Opt. 11, 34005 (2006).
[CrossRef] [PubMed]

Carl, D.

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schafer, W. Domschke, and G. von Bally, J. Biomed. Opt. 11, 34005 (2006).
[CrossRef] [PubMed]

Charriere, F.

Charrière, F.

Chaubal, K. A.

K. A. Chaubal, Z. Lodin, and J. Pilny, Acta Histochem. 26, 131 (1967).
[PubMed]

Colomb, T.

Cuche, E.

Curl, C. L.

C. L. Curl, C. J. Bellair, T. Harris, B. E. Allman, P. J. Harris, A. G. Stewart, A. Roberts, K. A. Nugent, and L. M. Delbridge, Cytometry 65, 88 (2005).
[CrossRef] [PubMed]

Dasari, R. R.

Delbridge, L. M.

C. L. Curl, C. J. Bellair, T. Harris, B. E. Allman, P. J. Harris, A. G. Stewart, A. Roberts, K. A. Nugent, and L. M. Delbridge, Cytometry 65, 88 (2005).
[CrossRef] [PubMed]

Depeursinge, C.

Depeursinge, Ch.

Domschke, W.

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schafer, W. Domschke, and G. von Bally, J. Biomed. Opt. 11, 34005 (2006).
[CrossRef] [PubMed]

Emery, Y.

Feld, M. S.

Harris, P. J.

C. L. Curl, C. J. Bellair, T. Harris, B. E. Allman, P. J. Harris, A. G. Stewart, A. Roberts, K. A. Nugent, and L. M. Delbridge, Cytometry 65, 88 (2005).
[CrossRef] [PubMed]

Harris, T.

C. L. Curl, C. J. Bellair, T. Harris, B. E. Allman, P. J. Harris, A. G. Stewart, A. Roberts, K. A. Nugent, and L. M. Delbridge, Cytometry 65, 88 (2005).
[CrossRef] [PubMed]

Ikeda, T.

Kemper, B.

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schafer, W. Domschke, and G. von Bally, J. Biomed. Opt. 11, 34005 (2006).
[CrossRef] [PubMed]

Kim, M.

Kuhn, J.

Kühn, J.

Lo, C.-M.

Lodin, Z.

K. A. Chaubal, Z. Lodin, and J. Pilny, Acta Histochem. 26, 131 (1967).
[PubMed]

Lucarini, V.

V. Lucarini, J. J. Saarinen, K.-E. Peiponen, and E. M. Vartiainen, Kramers-Kronig Relations in Optical Materials Research (Springer, 2005).

Lue, N.

Magistretti, P. J.

Mann, C.

Marian, A.

Marquet, P.

Montfort, F.

Nugent, K. A.

C. L. Curl, C. J. Bellair, T. Harris, B. E. Allman, P. J. Harris, A. G. Stewart, A. Roberts, K. A. Nugent, and L. M. Delbridge, Cytometry 65, 88 (2005).
[CrossRef] [PubMed]

Peiponen, K.-E.

V. Lucarini, J. J. Saarinen, K.-E. Peiponen, and E. M. Vartiainen, Kramers-Kronig Relations in Optical Materials Research (Springer, 2005).

Pilny, J.

K. A. Chaubal, Z. Lodin, and J. Pilny, Acta Histochem. 26, 131 (1967).
[PubMed]

Popescu, G.

Rappaz, B.

Roberts, A.

C. L. Curl, C. J. Bellair, T. Harris, B. E. Allman, P. J. Harris, A. G. Stewart, A. Roberts, K. A. Nugent, and L. M. Delbridge, Cytometry 65, 88 (2005).
[CrossRef] [PubMed]

Saarinen, J. J.

V. Lucarini, J. J. Saarinen, K.-E. Peiponen, and E. M. Vartiainen, Kramers-Kronig Relations in Optical Materials Research (Springer, 2005).

Schafer, M.

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schafer, W. Domschke, and G. von Bally, J. Biomed. Opt. 11, 34005 (2006).
[CrossRef] [PubMed]

Schnekenburger, J.

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schafer, W. Domschke, and G. von Bally, J. Biomed. Opt. 11, 34005 (2006).
[CrossRef] [PubMed]

Stewart, A. G.

C. L. Curl, C. J. Bellair, T. Harris, B. E. Allman, P. J. Harris, A. G. Stewart, A. Roberts, K. A. Nugent, and L. M. Delbridge, Cytometry 65, 88 (2005).
[CrossRef] [PubMed]

Vartiainen, E. M.

V. Lucarini, J. J. Saarinen, K.-E. Peiponen, and E. M. Vartiainen, Kramers-Kronig Relations in Optical Materials Research (Springer, 2005).

von Bally, G.

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schafer, W. Domschke, and G. von Bally, J. Biomed. Opt. 11, 34005 (2006).
[CrossRef] [PubMed]

Yu, L.

Acta Histochem. (1)

K. A. Chaubal, Z. Lodin, and J. Pilny, Acta Histochem. 26, 131 (1967).
[PubMed]

Cytometry (1)

C. L. Curl, C. J. Bellair, T. Harris, B. E. Allman, P. J. Harris, A. G. Stewart, A. Roberts, K. A. Nugent, and L. M. Delbridge, Cytometry 65, 88 (2005).
[CrossRef] [PubMed]

J. Biomed. Opt. (1)

B. Kemper, D. Carl, J. Schnekenburger, I. Bredebusch, M. Schafer, W. Domschke, and G. von Bally, J. Biomed. Opt. 11, 34005 (2006).
[CrossRef] [PubMed]

J. Histochem. Cytochem. (1)

R. Barer, J. Histochem. Cytochem. 17, 423 (1969).
[CrossRef] [PubMed]

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

Opt. Express (3)

Opt. Lett. (2)

Other (1)

V. Lucarini, J. J. Saarinen, K.-E. Peiponen, and E. M. Vartiainen, Kramers-Kronig Relations in Optical Materials Research (Springer, 2005).

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Figures (2)

Fig. 1
Fig. 1

Comparison of the KK prediction with experimental results. Dashed curve, measured absorption curve of SRB1; solid curve, RI variations prediction of the KK relations in the vicinity of the absorption peak; black diamonds, measured experimental RI shift. The Cauchy principal value has been evaluated relatively to a frequency ω 0 , corresponding to 663 nm , and n ( ω 0 ) has been arbitrarily set to zero.

Fig. 2
Fig. 2

(a) Phase image of two yeasts (S. Pombe) at 490 nm with measured regions highlighted in white, (b) and (c) variation of the optical-path difference for the two monitored cells (respectively, left and right cell) as a function of the wavelength.

Equations (3)

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n ( ω ) = n ( ω 0 ) + c π ( ω 2 ω 0 2 ) P 0 μ a ( ω ) ( ω 2 ω 2 ) ( ω 0 2 ω 2 ) d ω ,
OPD = ( n c n m ) d ,
n c 1 = OPD 1 ( Δ n m 1 , 2 OPD 2 OPD 1 ) + n m 1 ,

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