Abstract

Accurate refractive index measurement in the deep ultraviolet (UV) range is important for the separate quantification of biomolecules such as proteins and DNA in biology. This task is demanding and has not been fully exploited so far. Here we report a new method of measuring refractive index using field-based light scattering spectroscopy, which is applicable to any wavelength range and suitable for both solutions and homogenous objects with well-defined shape such as microspheres. The angular scattering distribution of single microspheres immersed in homogeneous media is measured over the wavelength range 260 to 315 nm using quantitative phase microscopy. By least square fitting the observed scattering distribution with Mie scattering theory, the refractive index of either the sphere or the immersion medium can be determined provided that one is known a priori. Using this method, we have measured the refractive index dispersion of SiO2 spheres and bovine serum albumin (BSA) solutions in the deep UV region. Specific refractive index increments of BSA are also extracted. Typical accuracy of the present refractive index technique is ≤0.003. The precision of refractive index measurements is ≤0.002 and that of specific refractive index increment determination is ≤0.01 mL/g.

© 2009 OSA

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2009

2008

W. Choi, C. C. Yu, C. Fang-Yen, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Field-based angle-resolved light-scattering study of single live cells,” Opt. Lett. 33(14), 1596–1598 (2008).
[CrossRef] [PubMed]

H. F. Ding, Z. Wang, F. Nguyen, S. A. Boppart, and G. Popescu, “Fourier transform light scattering of inhomogeneous and dynamic structures,” Phys. Rev. Lett. 101(23), 238102 (2008).
[CrossRef] [PubMed]

B. Rappaz, F. Charrière, C. Depeursinge, P. J. Magistretti, and P. Marquet, “Simultaneous cell morphometry and refractive index measurement with dual-wavelength digital holographic microscopy and dye-enhanced dispersion of perfusion medium,” Opt. Lett. 33(7), 744–746 (2008).
[CrossRef] [PubMed]

G. Popescu, Y. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, and S. Suresh, “Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum,” Proc. Natl. Acad. Sci. U.S.A. 105(37), 13730–13735 (2008).
[CrossRef] [PubMed]

2007

B. J. Zeskind, C. D. Jordan, W. Timp, L. Trapani, G. Waller, V. Horodincu, D. J. Ehrlich, and P. Matsudaira, “Nucleic acid and protein mass mapping by live-cell deep-ultraviolet microscopy,” Nat. Methods 4(7), 567–569 (2007).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007).
[CrossRef] [PubMed]

M. Daimon and A. Masumura, “Measurement of the refractive index of distilled water from the near-infrared region to the ultraviolet region,” Appl. Opt. 46(18), 3811–3820 (2007).
[CrossRef] [PubMed]

2006

2005

2004

A. Pinchuk, “Optical constants and dielectric function of DNA's nucleotides in UV range,” J. Quant. Spectrosc. Radiat. Transf. 85(2), 211–215 (2004).
[CrossRef]

2001

1998

1990

C. M. Stoscheck and P. D. Murray, “Quantitation of protein,” Methods Enzymol. 182, 50–68 (1990).
[CrossRef] [PubMed]

1974

M. Andersen, L. R. Painter, and S. Nir, “Dispersion-Equation and Polarizability of Bovine Serum-Albumin from Measurements of Refractive-Indexes,” Biopolymers 13(6), 1261–1267 (1974).
[CrossRef]

B. S. Chincholi, A. J. Havlik, and R. D. Vold, “Specific refractive index increments of polymer systems at four wavelengths,” J. Chem. Eng. Data 19(2), 148–152 (1974).
[CrossRef]

1957

Andersen, M.

M. Andersen, L. R. Painter, and S. Nir, “Dispersion-Equation and Polarizability of Bovine Serum-Albumin from Measurements of Refractive-Indexes,” Biopolymers 13(6), 1261–1267 (1974).
[CrossRef]

Badizadegan, K.

Y. J. Sung, W. Choi, C. Fang-Yen, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Optical diffraction tomography for high resolution live cell imaging,” Opt. Express 17(1), 266–277 (2009).
[CrossRef] [PubMed]

G. Popescu, Y. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

W. Choi, C. C. Yu, C. Fang-Yen, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Field-based angle-resolved light-scattering study of single live cells,” Opt. Lett. 33(14), 1596–1598 (2008).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007).
[CrossRef] [PubMed]

Barer, R.

Barty, A.

Best-Popescu, C.

G. Popescu, Y. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

Boppart, S. A.

H. F. Ding, Z. Wang, F. Nguyen, S. A. Boppart, and G. Popescu, “Fourier transform light scattering of inhomogeneous and dynamic structures,” Phys. Rev. Lett. 101(23), 238102 (2008).
[CrossRef] [PubMed]

Buddhiwant, P.

N. Ghosh, P. Buddhiwant, A. Uppal, S. K. Majumder, H. S. Patel, and P. K. Gupta, “Simultaneous determination of size and refractive index of red blood cells by light scattering measurements,” Appl. Phys. Lett. 88(8), 084101 (2006).
[CrossRef]

Charrière, F.

Chincholi, B. S.

B. S. Chincholi, A. J. Havlik, and R. D. Vold, “Specific refractive index increments of polymer systems at four wavelengths,” J. Chem. Eng. Data 19(2), 148–152 (1974).
[CrossRef]

Choi, W.

Y. J. Sung, W. Choi, C. Fang-Yen, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Optical diffraction tomography for high resolution live cell imaging,” Opt. Express 17(1), 266–277 (2009).
[CrossRef] [PubMed]

Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, and S. Suresh, “Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum,” Proc. Natl. Acad. Sci. U.S.A. 105(37), 13730–13735 (2008).
[CrossRef] [PubMed]

W. Choi, C. C. Yu, C. Fang-Yen, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Field-based angle-resolved light-scattering study of single live cells,” Opt. Lett. 33(14), 1596–1598 (2008).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007).
[CrossRef] [PubMed]

Colomb, T.

Cuche, E.

Daimon, M.

Dasari, R. R.

Deflores, L.

G. Popescu, Y. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

Depeursinge, C.

Diez-Silva, M.

Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, and S. Suresh, “Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum,” Proc. Natl. Acad. Sci. U.S.A. 105(37), 13730–13735 (2008).
[CrossRef] [PubMed]

Ding, H. F.

H. F. Ding, Z. Wang, F. Nguyen, S. A. Boppart, and G. Popescu, “Fourier transform light scattering of inhomogeneous and dynamic structures,” Phys. Rev. Lett. 101(23), 238102 (2008).
[CrossRef] [PubMed]

Ehrlich, D. J.

B. J. Zeskind, C. D. Jordan, W. Timp, L. Trapani, G. Waller, V. Horodincu, D. J. Ehrlich, and P. Matsudaira, “Nucleic acid and protein mass mapping by live-cell deep-ultraviolet microscopy,” Nat. Methods 4(7), 567–569 (2007).
[CrossRef] [PubMed]

Emery, Y.

Fang-Yen, C.

Feld, M. S.

Y. J. Sung, W. Choi, C. Fang-Yen, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Optical diffraction tomography for high resolution live cell imaging,” Opt. Express 17(1), 266–277 (2009).
[CrossRef] [PubMed]

Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, and S. Suresh, “Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum,” Proc. Natl. Acad. Sci. U.S.A. 105(37), 13730–13735 (2008).
[CrossRef] [PubMed]

G. Popescu, Y. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

W. Choi, C. C. Yu, C. Fang-Yen, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Field-based angle-resolved light-scattering study of single live cells,” Opt. Lett. 33(14), 1596–1598 (2008).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007).
[CrossRef] [PubMed]

G. Popescu, T. Ikeda, R. R. Dasari, and M. S. Feld, “Diffraction phase microscopy for quantifying cell structure and dynamics,” Opt. Lett. 31(6), 775–777 (2006).
[CrossRef] [PubMed]

T. Ikeda, G. Popescu, R. R. Dasari, and M. S. Feld, “Hilbert phase microscopy for investigating fast dynamics in transparent systems,” Opt. Lett. 30(10), 1165–1167 (2005).
[CrossRef] [PubMed]

A. Wax, C. Yang, R. R. Dasari, and M. S. Feld, “Measurement of angular distributions by use of low-coherence interferometry for light-scattering spectroscopy,” Opt. Lett. 26(6), 322–324 (2001).
[CrossRef]

Friebel, M.

M. Friebel and M. Meinke, “Determination of the complex refractive index of highly concentrated hemoglobin solutions using transmittance and reflectance measurements,” J. Biomed. Opt. 10(6), 064019–064015 (2005).
[CrossRef]

Ghosh, N.

N. Ghosh, P. Buddhiwant, A. Uppal, S. K. Majumder, H. S. Patel, and P. K. Gupta, “Simultaneous determination of size and refractive index of red blood cells by light scattering measurements,” Appl. Phys. Lett. 88(8), 084101 (2006).
[CrossRef]

Gupta, P. K.

N. Ghosh, P. Buddhiwant, A. Uppal, S. K. Majumder, H. S. Patel, and P. K. Gupta, “Simultaneous determination of size and refractive index of red blood cells by light scattering measurements,” Appl. Phys. Lett. 88(8), 084101 (2006).
[CrossRef]

Havlik, A. J.

B. S. Chincholi, A. J. Havlik, and R. D. Vold, “Specific refractive index increments of polymer systems at four wavelengths,” J. Chem. Eng. Data 19(2), 148–152 (1974).
[CrossRef]

Horodincu, V.

B. J. Zeskind, C. D. Jordan, W. Timp, L. Trapani, G. Waller, V. Horodincu, D. J. Ehrlich, and P. Matsudaira, “Nucleic acid and protein mass mapping by live-cell deep-ultraviolet microscopy,” Nat. Methods 4(7), 567–569 (2007).
[CrossRef] [PubMed]

Ikeda, T.

Jordan, C. D.

B. J. Zeskind, C. D. Jordan, W. Timp, L. Trapani, G. Waller, V. Horodincu, D. J. Ehrlich, and P. Matsudaira, “Nucleic acid and protein mass mapping by live-cell deep-ultraviolet microscopy,” Nat. Methods 4(7), 567–569 (2007).
[CrossRef] [PubMed]

Kim, M. K.

Kuehn, J.

Lo, C. M.

Lue, N.

G. Popescu, Y. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007).
[CrossRef] [PubMed]

Lykotrafitis, G.

Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, and S. Suresh, “Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum,” Proc. Natl. Acad. Sci. U.S.A. 105(37), 13730–13735 (2008).
[CrossRef] [PubMed]

Magistretti, P.

Magistretti, P. J.

Majumder, S. K.

N. Ghosh, P. Buddhiwant, A. Uppal, S. K. Majumder, H. S. Patel, and P. K. Gupta, “Simultaneous determination of size and refractive index of red blood cells by light scattering measurements,” Appl. Phys. Lett. 88(8), 084101 (2006).
[CrossRef]

Mann, C. J.

Marian, A.

Marquet, P.

Masumura, A.

Matsudaira, P.

B. J. Zeskind, C. D. Jordan, W. Timp, L. Trapani, G. Waller, V. Horodincu, D. J. Ehrlich, and P. Matsudaira, “Nucleic acid and protein mass mapping by live-cell deep-ultraviolet microscopy,” Nat. Methods 4(7), 567–569 (2007).
[CrossRef] [PubMed]

Meinke, M.

M. Friebel and M. Meinke, “Determination of the complex refractive index of highly concentrated hemoglobin solutions using transmittance and reflectance measurements,” J. Biomed. Opt. 10(6), 064019–064015 (2005).
[CrossRef]

Montfort, F.

Murray, P. D.

C. M. Stoscheck and P. D. Murray, “Quantitation of protein,” Methods Enzymol. 182, 50–68 (1990).
[CrossRef] [PubMed]

Nguyen, F.

H. F. Ding, Z. Wang, F. Nguyen, S. A. Boppart, and G. Popescu, “Fourier transform light scattering of inhomogeneous and dynamic structures,” Phys. Rev. Lett. 101(23), 238102 (2008).
[CrossRef] [PubMed]

Nir, S.

M. Andersen, L. R. Painter, and S. Nir, “Dispersion-Equation and Polarizability of Bovine Serum-Albumin from Measurements of Refractive-Indexes,” Biopolymers 13(6), 1261–1267 (1974).
[CrossRef]

Nugent, K. A.

Oh, S.

W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007).
[CrossRef] [PubMed]

Paganin, D.

Painter, L. R.

M. Andersen, L. R. Painter, and S. Nir, “Dispersion-Equation and Polarizability of Bovine Serum-Albumin from Measurements of Refractive-Indexes,” Biopolymers 13(6), 1261–1267 (1974).
[CrossRef]

Park, Y.

G. Popescu, Y. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, and S. Suresh, “Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum,” Proc. Natl. Acad. Sci. U.S.A. 105(37), 13730–13735 (2008).
[CrossRef] [PubMed]

Patel, H. S.

N. Ghosh, P. Buddhiwant, A. Uppal, S. K. Majumder, H. S. Patel, and P. K. Gupta, “Simultaneous determination of size and refractive index of red blood cells by light scattering measurements,” Appl. Phys. Lett. 88(8), 084101 (2006).
[CrossRef]

Pinchuk, A.

A. Pinchuk, “Optical constants and dielectric function of DNA's nucleotides in UV range,” J. Quant. Spectrosc. Radiat. Transf. 85(2), 211–215 (2004).
[CrossRef]

Popescu, G.

Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, and S. Suresh, “Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum,” Proc. Natl. Acad. Sci. U.S.A. 105(37), 13730–13735 (2008).
[CrossRef] [PubMed]

G. Popescu, Y. Park, N. Lue, C. Best-Popescu, L. Deflores, R. R. Dasari, M. S. Feld, and K. Badizadegan, “Optical imaging of cell mass and growth dynamics,” Am. J. Physiol. Cell Physiol. 295(2), C538–C544 (2008).
[CrossRef] [PubMed]

H. F. Ding, Z. Wang, F. Nguyen, S. A. Boppart, and G. Popescu, “Fourier transform light scattering of inhomogeneous and dynamic structures,” Phys. Rev. Lett. 101(23), 238102 (2008).
[CrossRef] [PubMed]

G. Popescu, T. Ikeda, R. R. Dasari, and M. S. Feld, “Diffraction phase microscopy for quantifying cell structure and dynamics,” Opt. Lett. 31(6), 775–777 (2006).
[CrossRef] [PubMed]

T. Ikeda, G. Popescu, R. R. Dasari, and M. S. Feld, “Hilbert phase microscopy for investigating fast dynamics in transparent systems,” Opt. Lett. 30(10), 1165–1167 (2005).
[CrossRef] [PubMed]

Rappaz, B.

Roberts, A.

Stoscheck, C. M.

C. M. Stoscheck and P. D. Murray, “Quantitation of protein,” Methods Enzymol. 182, 50–68 (1990).
[CrossRef] [PubMed]

Sung, Y. J.

Suresh, S.

Y. Park, M. Diez-Silva, G. Popescu, G. Lykotrafitis, W. Choi, M. S. Feld, and S. Suresh, “Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum,” Proc. Natl. Acad. Sci. U.S.A. 105(37), 13730–13735 (2008).
[CrossRef] [PubMed]

Timp, W.

B. J. Zeskind, C. D. Jordan, W. Timp, L. Trapani, G. Waller, V. Horodincu, D. J. Ehrlich, and P. Matsudaira, “Nucleic acid and protein mass mapping by live-cell deep-ultraviolet microscopy,” Nat. Methods 4(7), 567–569 (2007).
[CrossRef] [PubMed]

Trapani, L.

B. J. Zeskind, C. D. Jordan, W. Timp, L. Trapani, G. Waller, V. Horodincu, D. J. Ehrlich, and P. Matsudaira, “Nucleic acid and protein mass mapping by live-cell deep-ultraviolet microscopy,” Nat. Methods 4(7), 567–569 (2007).
[CrossRef] [PubMed]

Uppal, A.

N. Ghosh, P. Buddhiwant, A. Uppal, S. K. Majumder, H. S. Patel, and P. K. Gupta, “Simultaneous determination of size and refractive index of red blood cells by light scattering measurements,” Appl. Phys. Lett. 88(8), 084101 (2006).
[CrossRef]

Vold, R. D.

B. S. Chincholi, A. J. Havlik, and R. D. Vold, “Specific refractive index increments of polymer systems at four wavelengths,” J. Chem. Eng. Data 19(2), 148–152 (1974).
[CrossRef]

Waller, G.

B. J. Zeskind, C. D. Jordan, W. Timp, L. Trapani, G. Waller, V. Horodincu, D. J. Ehrlich, and P. Matsudaira, “Nucleic acid and protein mass mapping by live-cell deep-ultraviolet microscopy,” Nat. Methods 4(7), 567–569 (2007).
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[CrossRef] [PubMed]

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Figures (6)

Fig. 1
Fig. 1

Schematic of UV quantitative phase microscopy setup. OPA: optical parametric amplifier, BBO: β-Barium Borate, OBJ: objective, G: grating, IP1: image plane 1, FP: Fourier plane, SPF: spatial filter. IP2: image plane 2.

Fig. 2
Fig. 2

(a) interference image of a SiO2 sphere immersed in water and (b) background captured by the CCD at 315 nm; (c) normalized amplitude and (d) phase image of the same sphere; (e) intensity scattering pattern in logarithm scale derived from Fourier transform of the E-field in the image plane; (f) angular scattering spectrum obtained through azimuthal averaging of (e).

Fig. 3
Fig. 3

(a) Normalized 1/χ2 map of the difference between experimental scattering spectrum and that of Mie scattering as a function of sphere size and index of refraction. (b) Comparison of experimental scattering spectrum with the predicted scattering spectrum (Mie theory).

Fig. 4
Fig. 4

Comparison of refractive index increments over water determined using the FLSS method (red dots) at 633 nm to that measured by a commercial refractometer (black squares). The red and black lines are the corresponding linear fits of refractive index increments to BSA concentration; the slopes represent specific refractive index increments α. The error bars represent standard deviations.

Fig. 5
Fig. 5

Refractive index dispersion of SiO2 sphere determined by the FLSS method (black squares) and water obtained from reference [22] (red dots) in the UV range. The error bars for SiO2 sphere are the standard deviations of 6-8 measurements at each wavelength.

Fig. 6
Fig. 6

(a) Specific refractive index increments determined by the FLSS method at 315 nm through linear regression of refractive indices of protein solutions on protein concentrations. The error bar at each measurement point is the standard deviation of 6-8 measurements. (b) Refractive index dispersion of protein solution determined by the FLSS method (black squares) in the UV range. The error bar corresponds to linear regression error at each wavelength. Absorption of 0.25 g/mL BSA solution diluted by 400 times is also displayed (blue dots).

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