Abstract

The health of a eukaryotic cell depends on the proper functioning of its cell organelles. Characterizing these nanometer- to micrometer-scaled specialized subunits without disturbing the cell is challenging but can also provide valuable insights regarding the state of a cell. We show that objective-based scanning surface plasmon resonance microscopy can be used to analyze the refractive index of cell organelles quantitatively in a noninvasive and label-free manner with a lateral resolution at the diffraction limit.

© 2019 Optical Society of America

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  9. Y. Yang, H. Yu, X. Shan, W. Wang, X. Liu, S. Wang, and N. Tao, Small 11, 2878 (2015).
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  10. C.-T. Yang, R. Méjard, H. J. Griesser, P. O. Bagnaninchi, and B. Thierry, Anal. Chem. 87, 1456 (2015).
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  11. F. Zhang, S. Wang, L. Yin, Y. Yang, Y. Guan, W. Wang, H. Xu, and N. Tao, Anal. Chem. 87, 9960 (2015).
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  13. E. Kreysing, H. Hassani, N. Hampe, and A. Offenhäusser, ACS Nano 12, 8934 (2018).
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  15. S. Abuhattum, K. Kim, T. M. Franzmann, A. Eßlinger, D. Midtvedt, R. Schlüßler, S. Möllmert, H.-S. Kuan, S. Alberti, V. Zaburdaev, and J. Guck, Front. Phys. 6, 388876 (2018).
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  18. P. Müller, M. Schürmann, S. Girardo, G. Cojoc, and J. Guck, Opt. Express 26, 10729 (2018).
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    [Crossref]
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  25. P. Müller and J. Guck, J. Biophoton. 11, e201800095 (2018).
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  27. Z. A. Steelman, W. J. Eldridge, J. B. Weintraub, and A. Wax, J. Biophoton. 10, 1714 (2017).
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  29. M. P. Clausen, H. Colin-York, F. Schneider, C. Eggeling, and M. Fritzsche, J. Phys. D 50, 064002 (2017).
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2018 (7)

E. Kreysing, H. Hassani, N. Hampe, and A. Offenhäusser, ACS Nano 12, 8934 (2018).
[Crossref]

S. Abuhattum, K. Kim, T. M. Franzmann, A. Eßlinger, D. Midtvedt, R. Schlüßler, S. Möllmert, H.-S. Kuan, S. Alberti, V. Zaburdaev, and J. Guck, Front. Phys. 6, 388876 (2018).
[Crossref]

P. Müller, M. Schürmann, S. Girardo, G. Cojoc, and J. Guck, Opt. Express 26, 10729 (2018).
[Crossref]

G. Kim, M. Lee, S. Youn, E. Lee, D. Kwon, J. Shin, S. Lee, Y. S. Lee, and Y. Park, Sci. Rep. 8, 9192 (2018).
[Crossref]

M. A. Yurkin, J. Biophoton. 11, e201800033 (2018).
[Crossref]

P. Müller and J. Guck, J. Biophoton. 11, e201800095 (2018).
[Crossref]

Z. A. Steelman, W. J. Eldridge, and A. Wax, J. Biophoton. 11, e201800091 (2018).
[Crossref]

2017 (4)

Z. A. Steelman, W. J. Eldridge, J. B. Weintraub, and A. Wax, J. Biophoton. 10, 1714 (2017).
[Crossref]

K. P. Bohannon, R. W. Holz, and D. Axelrod, Microsc. Microanal. 23, 978 (2017).
[Crossref]

M. P. Clausen, H. Colin-York, F. Schneider, C. Eggeling, and M. Fritzsche, J. Phys. D 50, 064002 (2017).
[Crossref]

X.-W. Liu, Y. Yang, W. Wang, S. Wang, M. Gao, J. Wu, and N. Tao, Angew. Chem. Int. Ed. Engl. 56, 8855 (2017).
[Crossref]

2016 (1)

M. Schürmann, J. Scholze, P. Müller, J. Guck, and C. J. Chan, J. Biophoton. 9, 1068 (2016).
[Crossref]

2015 (4)

K. Syal, W. Wang, X. Shan, S. Wang, H.-Y. Chen, and N. Tao, Biosens. Bioelectron. 63, 131 (2015).
[Crossref]

Y. Yang, H. Yu, X. Shan, W. Wang, X. Liu, S. Wang, and N. Tao, Small 11, 2878 (2015).
[Crossref]

C.-T. Yang, R. Méjard, H. J. Griesser, P. O. Bagnaninchi, and B. Thierry, Anal. Chem. 87, 1456 (2015).
[Crossref]

F. Zhang, S. Wang, L. Yin, Y. Yang, Y. Guan, W. Wang, H. Xu, and N. Tao, Anal. Chem. 87, 9960 (2015).
[Crossref]

2014 (2)

K. Toma, H. Kano, and A. Offenhäusser, ACS Nano 8, 12612 (2014).
[Crossref]

A. W. Peterson, M. Halter, A. Tona, and A. L. Plant, BMC Cell Biol. 15, 35 (2014).
[Crossref]

2013 (2)

A. E. Ekpenyong, S. M. Man, S. Achouri, C. E. Bryant, J. Guck, and K. J. Chalut, J. Biophoton. 6, 393 (2013).
[Crossref]

A. G. Clark, K. Dierkes, and E. K. Paluch, Biophys. J. 105, 570 (2013).
[Crossref]

2012 (2)

W. Wang, Y. Yang, S. Wang, V. J. Nagaraj, Q. Liu, J. Wu, and N. Tao, Nat. Chem. 4, 846 (2012).
[Crossref]

K. Watanabe, K. Matsuura, F. Kawata, K. Nagata, J. Ning, and H. Kano, Biomed. Opt. Express 3, 354 (2012).
[Crossref]

2009 (1)

2008 (1)

2007 (2)

B. Huang, F. Yu, and R. N. Zare, Anal. Chem. 79, 2979 (2007).
[Crossref]

K. Watanabe, N. Horiguchi, and H. Kano, Appl. Opt. 46, 4985(2007).
[Crossref]

1999 (1)

K.-F. Giebel, C. Bechinger, S. Herminghaus, M. Riedel, P. Leiderer, U. Weiland, and M. Bastmeyer, Biophys. J. 76, 509 (1999).
[Crossref]

Abuhattum, S.

S. Abuhattum, K. Kim, T. M. Franzmann, A. Eßlinger, D. Midtvedt, R. Schlüßler, S. Möllmert, H.-S. Kuan, S. Alberti, V. Zaburdaev, and J. Guck, Front. Phys. 6, 388876 (2018).
[Crossref]

Achouri, S.

A. E. Ekpenyong, S. M. Man, S. Achouri, C. E. Bryant, J. Guck, and K. J. Chalut, J. Biophoton. 6, 393 (2013).
[Crossref]

Ae Kim, S.

Albert Kim, D.-G.

Alberti, S.

S. Abuhattum, K. Kim, T. M. Franzmann, A. Eßlinger, D. Midtvedt, R. Schlüßler, S. Möllmert, H.-S. Kuan, S. Alberti, V. Zaburdaev, and J. Guck, Front. Phys. 6, 388876 (2018).
[Crossref]

Axelrod, D.

K. P. Bohannon, R. W. Holz, and D. Axelrod, Microsc. Microanal. 23, 978 (2017).
[Crossref]

Baac, H.

Badizadegan, K.

Bagnaninchi, P. O.

C.-T. Yang, R. Méjard, H. J. Griesser, P. O. Bagnaninchi, and B. Thierry, Anal. Chem. 87, 1456 (2015).
[Crossref]

Bass, M.

M. Bass, C. DeCusatis, J. M. Enoch, V. Lakshminarayanan, G. Li, C. MacDonald, and V. N. Mahajan, Handbook of Optics (McGraw-Hill, 2010).

Bastmeyer, M.

K.-F. Giebel, C. Bechinger, S. Herminghaus, M. Riedel, P. Leiderer, U. Weiland, and M. Bastmeyer, Biophys. J. 76, 509 (1999).
[Crossref]

Bechinger, C.

K.-F. Giebel, C. Bechinger, S. Herminghaus, M. Riedel, P. Leiderer, U. Weiland, and M. Bastmeyer, Biophys. J. 76, 509 (1999).
[Crossref]

Bohannon, K. P.

K. P. Bohannon, R. W. Holz, and D. Axelrod, Microsc. Microanal. 23, 978 (2017).
[Crossref]

Bryant, C. E.

A. E. Ekpenyong, S. M. Man, S. Achouri, C. E. Bryant, J. Guck, and K. J. Chalut, J. Biophoton. 6, 393 (2013).
[Crossref]

Chalut, K. J.

A. E. Ekpenyong, S. M. Man, S. Achouri, C. E. Bryant, J. Guck, and K. J. Chalut, J. Biophoton. 6, 393 (2013).
[Crossref]

M. Schürmann, J. Scholze, P. Müller, C. J. Chan, A. E. Ekpenyong, K. J. Chalut, and J. Guck, in Methods in Cell Biology, E. K. Paluch, ed., Vol. 125 of Biophysical Methods in Cell Biology (Academic, 2015), Chap. 9, pp. 143.

Chan, C. J.

M. Schürmann, J. Scholze, P. Müller, J. Guck, and C. J. Chan, J. Biophoton. 9, 1068 (2016).
[Crossref]

M. Schürmann, J. Scholze, P. Müller, C. J. Chan, A. E. Ekpenyong, K. J. Chalut, and J. Guck, in Methods in Cell Biology, E. K. Paluch, ed., Vol. 125 of Biophysical Methods in Cell Biology (Academic, 2015), Chap. 9, pp. 143.

Chen, H.-Y.

K. Syal, W. Wang, X. Shan, S. Wang, H.-Y. Chen, and N. Tao, Biosens. Bioelectron. 63, 131 (2015).
[Crossref]

Choi, W.

Clark, A. G.

A. G. Clark, K. Dierkes, and E. K. Paluch, Biophys. J. 105, 570 (2013).
[Crossref]

Clausen, M. P.

M. P. Clausen, H. Colin-York, F. Schneider, C. Eggeling, and M. Fritzsche, J. Phys. D 50, 064002 (2017).
[Crossref]

Cojoc, G.

Colin-York, H.

M. P. Clausen, H. Colin-York, F. Schneider, C. Eggeling, and M. Fritzsche, J. Phys. D 50, 064002 (2017).
[Crossref]

Dasari, R. R.

DeCusatis, C.

M. Bass, C. DeCusatis, J. M. Enoch, V. Lakshminarayanan, G. Li, C. MacDonald, and V. N. Mahajan, Handbook of Optics (McGraw-Hill, 2010).

Dierkes, K.

A. G. Clark, K. Dierkes, and E. K. Paluch, Biophys. J. 105, 570 (2013).
[Crossref]

Eggeling, C.

M. P. Clausen, H. Colin-York, F. Schneider, C. Eggeling, and M. Fritzsche, J. Phys. D 50, 064002 (2017).
[Crossref]

Ekpenyong, A. E.

A. E. Ekpenyong, S. M. Man, S. Achouri, C. E. Bryant, J. Guck, and K. J. Chalut, J. Biophoton. 6, 393 (2013).
[Crossref]

M. Schürmann, J. Scholze, P. Müller, C. J. Chan, A. E. Ekpenyong, K. J. Chalut, and J. Guck, in Methods in Cell Biology, E. K. Paluch, ed., Vol. 125 of Biophysical Methods in Cell Biology (Academic, 2015), Chap. 9, pp. 143.

Eldridge, W. J.

Z. A. Steelman, W. J. Eldridge, and A. Wax, J. Biophoton. 11, e201800091 (2018).
[Crossref]

Z. A. Steelman, W. J. Eldridge, J. B. Weintraub, and A. Wax, J. Biophoton. 10, 1714 (2017).
[Crossref]

Enoch, J. M.

M. Bass, C. DeCusatis, J. M. Enoch, V. Lakshminarayanan, G. Li, C. MacDonald, and V. N. Mahajan, Handbook of Optics (McGraw-Hill, 2010).

Eßlinger, A.

S. Abuhattum, K. Kim, T. M. Franzmann, A. Eßlinger, D. Midtvedt, R. Schlüßler, S. Möllmert, H.-S. Kuan, S. Alberti, V. Zaburdaev, and J. Guck, Front. Phys. 6, 388876 (2018).
[Crossref]

Fang-Yen, C.

Feld, M. S.

Franzmann, T. M.

S. Abuhattum, K. Kim, T. M. Franzmann, A. Eßlinger, D. Midtvedt, R. Schlüßler, S. Möllmert, H.-S. Kuan, S. Alberti, V. Zaburdaev, and J. Guck, Front. Phys. 6, 388876 (2018).
[Crossref]

Fritzsche, M.

M. P. Clausen, H. Colin-York, F. Schneider, C. Eggeling, and M. Fritzsche, J. Phys. D 50, 064002 (2017).
[Crossref]

Gao, M.

X.-W. Liu, Y. Yang, W. Wang, S. Wang, M. Gao, J. Wu, and N. Tao, Angew. Chem. Int. Ed. Engl. 56, 8855 (2017).
[Crossref]

Giebel, K.-F.

K.-F. Giebel, C. Bechinger, S. Herminghaus, M. Riedel, P. Leiderer, U. Weiland, and M. Bastmeyer, Biophys. J. 76, 509 (1999).
[Crossref]

Girardo, S.

Griesser, H. J.

C.-T. Yang, R. Méjard, H. J. Griesser, P. O. Bagnaninchi, and B. Thierry, Anal. Chem. 87, 1456 (2015).
[Crossref]

Guan, Y.

F. Zhang, S. Wang, L. Yin, Y. Yang, Y. Guan, W. Wang, H. Xu, and N. Tao, Anal. Chem. 87, 9960 (2015).
[Crossref]

Guck, J.

S. Abuhattum, K. Kim, T. M. Franzmann, A. Eßlinger, D. Midtvedt, R. Schlüßler, S. Möllmert, H.-S. Kuan, S. Alberti, V. Zaburdaev, and J. Guck, Front. Phys. 6, 388876 (2018).
[Crossref]

P. Müller and J. Guck, J. Biophoton. 11, e201800095 (2018).
[Crossref]

P. Müller, M. Schürmann, S. Girardo, G. Cojoc, and J. Guck, Opt. Express 26, 10729 (2018).
[Crossref]

M. Schürmann, J. Scholze, P. Müller, J. Guck, and C. J. Chan, J. Biophoton. 9, 1068 (2016).
[Crossref]

A. E. Ekpenyong, S. M. Man, S. Achouri, C. E. Bryant, J. Guck, and K. J. Chalut, J. Biophoton. 6, 393 (2013).
[Crossref]

M. Schürmann, J. Scholze, P. Müller, C. J. Chan, A. E. Ekpenyong, K. J. Chalut, and J. Guck, in Methods in Cell Biology, E. K. Paluch, ed., Vol. 125 of Biophysical Methods in Cell Biology (Academic, 2015), Chap. 9, pp. 143.

Halter, M.

A. W. Peterson, M. Halter, A. Tona, and A. L. Plant, BMC Cell Biol. 15, 35 (2014).
[Crossref]

Hampe, N.

E. Kreysing, H. Hassani, N. Hampe, and A. Offenhäusser, ACS Nano 12, 8934 (2018).
[Crossref]

Hassani, H.

E. Kreysing, H. Hassani, N. Hampe, and A. Offenhäusser, ACS Nano 12, 8934 (2018).
[Crossref]

Herminghaus, S.

K.-F. Giebel, C. Bechinger, S. Herminghaus, M. Riedel, P. Leiderer, U. Weiland, and M. Bastmeyer, Biophys. J. 76, 509 (1999).
[Crossref]

Holz, R. W.

K. P. Bohannon, R. W. Holz, and D. Axelrod, Microsc. Microanal. 23, 978 (2017).
[Crossref]

Hoon Kim, J.

Horiguchi, N.

Huang, B.

B. Huang, F. Yu, and R. N. Zare, Anal. Chem. 79, 2979 (2007).
[Crossref]

Kano, H.

Kawata, F.

Kim, G.

G. Kim, M. Lee, S. Youn, E. Lee, D. Kwon, J. Shin, S. Lee, Y. S. Lee, and Y. Park, Sci. Rep. 8, 9192 (2018).
[Crossref]

Kim, K.

S. Abuhattum, K. Kim, T. M. Franzmann, A. Eßlinger, D. Midtvedt, R. Schlüßler, S. Möllmert, H.-S. Kuan, S. Alberti, V. Zaburdaev, and J. Guck, Front. Phys. 6, 388876 (2018).
[Crossref]

Kim, S. J.

Kreysing, E.

E. Kreysing, H. Hassani, N. Hampe, and A. Offenhäusser, ACS Nano 12, 8934 (2018).
[Crossref]

Kuan, H.-S.

S. Abuhattum, K. Kim, T. M. Franzmann, A. Eßlinger, D. Midtvedt, R. Schlüßler, S. Möllmert, H.-S. Kuan, S. Alberti, V. Zaburdaev, and J. Guck, Front. Phys. 6, 388876 (2018).
[Crossref]

Kwon, D.

G. Kim, M. Lee, S. Youn, E. Lee, D. Kwon, J. Shin, S. Lee, Y. S. Lee, and Y. Park, Sci. Rep. 8, 9192 (2018).
[Crossref]

Lakshminarayanan, V.

M. Bass, C. DeCusatis, J. M. Enoch, V. Lakshminarayanan, G. Li, C. MacDonald, and V. N. Mahajan, Handbook of Optics (McGraw-Hill, 2010).

Lee, E.

G. Kim, M. Lee, S. Youn, E. Lee, D. Kwon, J. Shin, S. Lee, Y. S. Lee, and Y. Park, Sci. Rep. 8, 9192 (2018).
[Crossref]

Lee, J.

Lee, M.

G. Kim, M. Lee, S. Youn, E. Lee, D. Kwon, J. Shin, S. Lee, Y. S. Lee, and Y. Park, Sci. Rep. 8, 9192 (2018).
[Crossref]

Lee, S.

G. Kim, M. Lee, S. Youn, E. Lee, D. Kwon, J. Shin, S. Lee, Y. S. Lee, and Y. Park, Sci. Rep. 8, 9192 (2018).
[Crossref]

Lee, Y. S.

G. Kim, M. Lee, S. Youn, E. Lee, D. Kwon, J. Shin, S. Lee, Y. S. Lee, and Y. Park, Sci. Rep. 8, 9192 (2018).
[Crossref]

Leiderer, P.

K.-F. Giebel, C. Bechinger, S. Herminghaus, M. Riedel, P. Leiderer, U. Weiland, and M. Bastmeyer, Biophys. J. 76, 509 (1999).
[Crossref]

Li, G.

M. Bass, C. DeCusatis, J. M. Enoch, V. Lakshminarayanan, G. Li, C. MacDonald, and V. N. Mahajan, Handbook of Optics (McGraw-Hill, 2010).

Liu, Q.

W. Wang, Y. Yang, S. Wang, V. J. Nagaraj, Q. Liu, J. Wu, and N. Tao, Nat. Chem. 4, 846 (2012).
[Crossref]

Liu, X.

Y. Yang, H. Yu, X. Shan, W. Wang, X. Liu, S. Wang, and N. Tao, Small 11, 2878 (2015).
[Crossref]

Liu, X.-W.

X.-W. Liu, Y. Yang, W. Wang, S. Wang, M. Gao, J. Wu, and N. Tao, Angew. Chem. Int. Ed. Engl. 56, 8855 (2017).
[Crossref]

MacDonald, C.

M. Bass, C. DeCusatis, J. M. Enoch, V. Lakshminarayanan, G. Li, C. MacDonald, and V. N. Mahajan, Handbook of Optics (McGraw-Hill, 2010).

Mahajan, V. N.

M. Bass, C. DeCusatis, J. M. Enoch, V. Lakshminarayanan, G. Li, C. MacDonald, and V. N. Mahajan, Handbook of Optics (McGraw-Hill, 2010).

Man, S. M.

A. E. Ekpenyong, S. M. Man, S. Achouri, C. E. Bryant, J. Guck, and K. J. Chalut, J. Biophoton. 6, 393 (2013).
[Crossref]

Matsuura, K.

Méjard, R.

C.-T. Yang, R. Méjard, H. J. Griesser, P. O. Bagnaninchi, and B. Thierry, Anal. Chem. 87, 1456 (2015).
[Crossref]

Midtvedt, D.

S. Abuhattum, K. Kim, T. M. Franzmann, A. Eßlinger, D. Midtvedt, R. Schlüßler, S. Möllmert, H.-S. Kuan, S. Alberti, V. Zaburdaev, and J. Guck, Front. Phys. 6, 388876 (2018).
[Crossref]

Min Byun, K.

Möllmert, S.

S. Abuhattum, K. Kim, T. M. Franzmann, A. Eßlinger, D. Midtvedt, R. Schlüßler, S. Möllmert, H.-S. Kuan, S. Alberti, V. Zaburdaev, and J. Guck, Front. Phys. 6, 388876 (2018).
[Crossref]

Müller, P.

P. Müller and J. Guck, J. Biophoton. 11, e201800095 (2018).
[Crossref]

P. Müller, M. Schürmann, S. Girardo, G. Cojoc, and J. Guck, Opt. Express 26, 10729 (2018).
[Crossref]

M. Schürmann, J. Scholze, P. Müller, J. Guck, and C. J. Chan, J. Biophoton. 9, 1068 (2016).
[Crossref]

M. Schürmann, J. Scholze, P. Müller, C. J. Chan, A. E. Ekpenyong, K. J. Chalut, and J. Guck, in Methods in Cell Biology, E. K. Paluch, ed., Vol. 125 of Biophysical Methods in Cell Biology (Academic, 2015), Chap. 9, pp. 143.

Nagaraj, V. J.

W. Wang, Y. Yang, S. Wang, V. J. Nagaraj, Q. Liu, J. Wu, and N. Tao, Nat. Chem. 4, 846 (2012).
[Crossref]

Nagata, K.

Ning, J.

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B. Huang, F. Yu, and R. N. Zare, Anal. Chem. 79, 2979 (2007).
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K. Toma, H. Kano, and A. Offenhäusser, ACS Nano 8, 12612 (2014).
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F. Zhang, S. Wang, L. Yin, Y. Yang, Y. Guan, W. Wang, H. Xu, and N. Tao, Anal. Chem. 87, 9960 (2015).
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B. Huang, F. Yu, and R. N. Zare, Anal. Chem. 79, 2979 (2007).
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Angew. Chem. Int. Ed. Engl. (1)

X.-W. Liu, Y. Yang, W. Wang, S. Wang, M. Gao, J. Wu, and N. Tao, Angew. Chem. Int. Ed. Engl. 56, 8855 (2017).
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Appl. Opt. (1)

Biomed. Opt. Express (1)

Biophys. J. (2)

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K. Syal, W. Wang, X. Shan, S. Wang, H.-Y. Chen, and N. Tao, Biosens. Bioelectron. 63, 131 (2015).
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BMC Cell Biol. (1)

A. W. Peterson, M. Halter, A. Tona, and A. L. Plant, BMC Cell Biol. 15, 35 (2014).
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Front. Phys. (1)

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J. Biophoton. (6)

M. A. Yurkin, J. Biophoton. 11, e201800033 (2018).
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P. Müller and J. Guck, J. Biophoton. 11, e201800095 (2018).
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J. Phys. D (1)

M. P. Clausen, H. Colin-York, F. Schneider, C. Eggeling, and M. Fritzsche, J. Phys. D 50, 064002 (2017).
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K. P. Bohannon, R. W. Holz, and D. Axelrod, Microsc. Microanal. 23, 978 (2017).
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Nat. Chem. (1)

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Opt. Express (2)

Opt. Lett. (1)

Sci. Rep. (1)

G. Kim, M. Lee, S. Youn, E. Lee, D. Kwon, J. Shin, S. Lee, Y. S. Lee, and Y. Park, Sci. Rep. 8, 9192 (2018).
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Small (1)

Y. Yang, H. Yu, X. Shan, W. Wang, X. Liu, S. Wang, and N. Tao, Small 11, 2878 (2015).
[Crossref]

Other (2)

M. Schürmann, J. Scholze, P. Müller, C. J. Chan, A. E. Ekpenyong, K. J. Chalut, and J. Guck, in Methods in Cell Biology, E. K. Paluch, ed., Vol. 125 of Biophysical Methods in Cell Biology (Academic, 2015), Chap. 9, pp. 143.

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Figures (3)

Fig. 1.
Fig. 1. (a) By illuminating the TIRF objective with radially polarized light, the light is focused spot-wise at the glass–gold interface. (b) Light arriving at the interface at angles higher than the critical angle θc creates an evanescent field that penetrates the cytosol and interacts with cell organelles. (c) The radial polarization is realized by a phase plate consisting of four elements. Lying in a conjugated plane, their intersections appear as a black cross when imaging the BFP. Since illumination with radially polarized light is only guaranteed by light passing through the centers of the four quarters (marked by white frame), we analyze only these central parts of the BFP images. (d) The resulting reflectance curve is analyzed by fitting a multilayer system to the data. The position of θc, represented by a prominent edge in the curve, depends only on the RI of the first and the last material within the penetration depth and can be used to determine the RI of cell organelles in close vicinity of the cell membrane.

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Fig. 2.
Fig. 2. Correlating a (a) phase contrast image of a neuron with an (b) SPRM analysis of the RI profile of the cell reveal that the RI of the nucleus is lower than the surrounding cytosol. Correlating a bright-field image of a neuron with its RI profile demonstrates that even the RI of very small cell organelles can be measured with SPRM. (e) The dashed lines indicate the positions of two individual scan lines. (f) and (g) The intracellular RI along the scan lines reveals the RI of the cell organelles D and B, confirming the high lateral resolution.

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Fig. 3.
Fig. 3. Noise around the critical angle affects the fit quality and therefore the accuracy in the RI measurement. (a) The reflectance profile at a scanning point with low noise allows for a high accuracy in RI. (b) The relatively high noise around the critical angle at this scanning point leads to an underestimation of the critical angle and therefore to an underestimation of the RI. Using a high-quality radial polarizer is likely to reduce the noise in the reflectance curve.

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Tables (1)

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Table 1. Multilayer System for a Sample Used in Cell Culture

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