Abstract

We present a practical modification of fiber-coupled confocal Raman scanning microscopes that is able to provide high confocal resolution in conjunction with high light collection efficiency. For this purpose, the single detection fiber is replaced by a hexagonal lenslet array in combination with a hexagonally packed round-to-linear multimode fiber bundle. A multiline detector is used to collect individual Raman spectra for each fiber. Data post-processing based on pixel reassignment allows one to improve the lateral resolution by up to 41% compared to a single fiber of equal light collection efficiency. We present results from an experimental implementation featuring seven collection fibers, yielding a resolution improvement of about 30%. We believe that our implementation represents an attractive upgrade for existing confocal Raman microscopes that employ multi-line detectors.

© 2016 Optical Society of America

Full Article  |  PDF Article
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References

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  1. M. Lamy de la Chapelle, P. Gucciardi, and N. Lidgi-Guigui, eds., Handbook of Enhanced Spectroscopy (Pan Stanford Publishing Pte Ltd., 2015).
  2. T. Deckert-Gaudig and V. Deckert, Phys. Chem. Chem. Phys. 12, 12040 (2010).
    [Crossref]
  3. K. Watanabe, A. Palonpon, N. Smith, L. Chiu, A. Kasai, H. Hashimoto, S. Kawata, and K. Fujita, Nat. Commun. 6, 10095 (2015).
    [Crossref]
  4. C. J. R. Sheppard, Optik 80, 0030 (1988).
  5. C. Müller and J. Enderlein, Phys. Rev. Lett. 104, 198101 (2010).
    [Crossref]
  6. A. G. York, S. H. Parekh, D. Dalle Nogare, R. S. Fischer, K. Temprine, M. Mione, A. B. Chitnis, C. A. Combs, and H. Shroff, Nat. Methods 9, 749 (2012).
    [Crossref]
  7. O. Schulz, C. Pieper, M. Clever, J. Pfaff, A. Ruhlandt, R. H. Kehlenbach, F. S. Wouters, J. Großhans, G. Bunt, and J. Enderlein, Proc. Natl. Acad. Sci. USA 110, 21000 (2013).
    [Crossref]
  8. M. Ingaramo, A. G. York, P. Wawrzusin, O. Milberg, A. Hong, R. Weigert, H. Shroff, and G. H. Patterson, Proc. Natl. Acad. Sci. USA 111, 5254 (2014).
    [Crossref]
  9. A. G. York, P. Chandris, D. Dalle Nogare, J. Head, P. Wawrzusin, R. S. Fischer, A. Chitnis, and H. Shroff, Nat. Methods 10, 1122 (2013).
    [Crossref]
  10. S. Roth, C. J. R. Sheppard, K. Wicker, and R. Heintzmann, Opt. Nanosc. 2, 5 (2013).
    [Crossref]
  11. G. M. R. De Luca, R. M. P. Breedijk, R. A. J. Brandt, C. H. C. Zeelenberg, B. E. de Jong, W. Timmermans, L. Nahidi Azar, R. A. Hoebe, S. Stallinga, and E. M. M. Manders, Biomed. Opt. Express 4, 2644 (2013).
    [Crossref]
  12. N. Hagen and M. W. Kudenov, Opt. Eng. 52, 090901 (2013).
    [Crossref]
  13. S. Roth, C. J. R. Sheppard, and R. Heintzmann, Opt. Lett. 41, 2109 (2016).
    [Crossref]

2016 (1)

2015 (1)

K. Watanabe, A. Palonpon, N. Smith, L. Chiu, A. Kasai, H. Hashimoto, S. Kawata, and K. Fujita, Nat. Commun. 6, 10095 (2015).
[Crossref]

2014 (1)

M. Ingaramo, A. G. York, P. Wawrzusin, O. Milberg, A. Hong, R. Weigert, H. Shroff, and G. H. Patterson, Proc. Natl. Acad. Sci. USA 111, 5254 (2014).
[Crossref]

2013 (5)

A. G. York, P. Chandris, D. Dalle Nogare, J. Head, P. Wawrzusin, R. S. Fischer, A. Chitnis, and H. Shroff, Nat. Methods 10, 1122 (2013).
[Crossref]

S. Roth, C. J. R. Sheppard, K. Wicker, and R. Heintzmann, Opt. Nanosc. 2, 5 (2013).
[Crossref]

N. Hagen and M. W. Kudenov, Opt. Eng. 52, 090901 (2013).
[Crossref]

G. M. R. De Luca, R. M. P. Breedijk, R. A. J. Brandt, C. H. C. Zeelenberg, B. E. de Jong, W. Timmermans, L. Nahidi Azar, R. A. Hoebe, S. Stallinga, and E. M. M. Manders, Biomed. Opt. Express 4, 2644 (2013).
[Crossref]

O. Schulz, C. Pieper, M. Clever, J. Pfaff, A. Ruhlandt, R. H. Kehlenbach, F. S. Wouters, J. Großhans, G. Bunt, and J. Enderlein, Proc. Natl. Acad. Sci. USA 110, 21000 (2013).
[Crossref]

2012 (1)

A. G. York, S. H. Parekh, D. Dalle Nogare, R. S. Fischer, K. Temprine, M. Mione, A. B. Chitnis, C. A. Combs, and H. Shroff, Nat. Methods 9, 749 (2012).
[Crossref]

2010 (2)

T. Deckert-Gaudig and V. Deckert, Phys. Chem. Chem. Phys. 12, 12040 (2010).
[Crossref]

C. Müller and J. Enderlein, Phys. Rev. Lett. 104, 198101 (2010).
[Crossref]

1988 (1)

C. J. R. Sheppard, Optik 80, 0030 (1988).

Brandt, R. A. J.

Breedijk, R. M. P.

Bunt, G.

O. Schulz, C. Pieper, M. Clever, J. Pfaff, A. Ruhlandt, R. H. Kehlenbach, F. S. Wouters, J. Großhans, G. Bunt, and J. Enderlein, Proc. Natl. Acad. Sci. USA 110, 21000 (2013).
[Crossref]

Chandris, P.

A. G. York, P. Chandris, D. Dalle Nogare, J. Head, P. Wawrzusin, R. S. Fischer, A. Chitnis, and H. Shroff, Nat. Methods 10, 1122 (2013).
[Crossref]

Chitnis, A.

A. G. York, P. Chandris, D. Dalle Nogare, J. Head, P. Wawrzusin, R. S. Fischer, A. Chitnis, and H. Shroff, Nat. Methods 10, 1122 (2013).
[Crossref]

Chitnis, A. B.

A. G. York, S. H. Parekh, D. Dalle Nogare, R. S. Fischer, K. Temprine, M. Mione, A. B. Chitnis, C. A. Combs, and H. Shroff, Nat. Methods 9, 749 (2012).
[Crossref]

Chiu, L.

K. Watanabe, A. Palonpon, N. Smith, L. Chiu, A. Kasai, H. Hashimoto, S. Kawata, and K. Fujita, Nat. Commun. 6, 10095 (2015).
[Crossref]

Clever, M.

O. Schulz, C. Pieper, M. Clever, J. Pfaff, A. Ruhlandt, R. H. Kehlenbach, F. S. Wouters, J. Großhans, G. Bunt, and J. Enderlein, Proc. Natl. Acad. Sci. USA 110, 21000 (2013).
[Crossref]

Combs, C. A.

A. G. York, S. H. Parekh, D. Dalle Nogare, R. S. Fischer, K. Temprine, M. Mione, A. B. Chitnis, C. A. Combs, and H. Shroff, Nat. Methods 9, 749 (2012).
[Crossref]

Dalle Nogare, D.

A. G. York, P. Chandris, D. Dalle Nogare, J. Head, P. Wawrzusin, R. S. Fischer, A. Chitnis, and H. Shroff, Nat. Methods 10, 1122 (2013).
[Crossref]

A. G. York, S. H. Parekh, D. Dalle Nogare, R. S. Fischer, K. Temprine, M. Mione, A. B. Chitnis, C. A. Combs, and H. Shroff, Nat. Methods 9, 749 (2012).
[Crossref]

de Jong, B. E.

De Luca, G. M. R.

Deckert, V.

T. Deckert-Gaudig and V. Deckert, Phys. Chem. Chem. Phys. 12, 12040 (2010).
[Crossref]

Deckert-Gaudig, T.

T. Deckert-Gaudig and V. Deckert, Phys. Chem. Chem. Phys. 12, 12040 (2010).
[Crossref]

Enderlein, J.

O. Schulz, C. Pieper, M. Clever, J. Pfaff, A. Ruhlandt, R. H. Kehlenbach, F. S. Wouters, J. Großhans, G. Bunt, and J. Enderlein, Proc. Natl. Acad. Sci. USA 110, 21000 (2013).
[Crossref]

C. Müller and J. Enderlein, Phys. Rev. Lett. 104, 198101 (2010).
[Crossref]

Fischer, R. S.

A. G. York, P. Chandris, D. Dalle Nogare, J. Head, P. Wawrzusin, R. S. Fischer, A. Chitnis, and H. Shroff, Nat. Methods 10, 1122 (2013).
[Crossref]

A. G. York, S. H. Parekh, D. Dalle Nogare, R. S. Fischer, K. Temprine, M. Mione, A. B. Chitnis, C. A. Combs, and H. Shroff, Nat. Methods 9, 749 (2012).
[Crossref]

Fujita, K.

K. Watanabe, A. Palonpon, N. Smith, L. Chiu, A. Kasai, H. Hashimoto, S. Kawata, and K. Fujita, Nat. Commun. 6, 10095 (2015).
[Crossref]

Großhans, J.

O. Schulz, C. Pieper, M. Clever, J. Pfaff, A. Ruhlandt, R. H. Kehlenbach, F. S. Wouters, J. Großhans, G. Bunt, and J. Enderlein, Proc. Natl. Acad. Sci. USA 110, 21000 (2013).
[Crossref]

Hagen, N.

N. Hagen and M. W. Kudenov, Opt. Eng. 52, 090901 (2013).
[Crossref]

Hashimoto, H.

K. Watanabe, A. Palonpon, N. Smith, L. Chiu, A. Kasai, H. Hashimoto, S. Kawata, and K. Fujita, Nat. Commun. 6, 10095 (2015).
[Crossref]

Head, J.

A. G. York, P. Chandris, D. Dalle Nogare, J. Head, P. Wawrzusin, R. S. Fischer, A. Chitnis, and H. Shroff, Nat. Methods 10, 1122 (2013).
[Crossref]

Heintzmann, R.

S. Roth, C. J. R. Sheppard, and R. Heintzmann, Opt. Lett. 41, 2109 (2016).
[Crossref]

S. Roth, C. J. R. Sheppard, K. Wicker, and R. Heintzmann, Opt. Nanosc. 2, 5 (2013).
[Crossref]

Hoebe, R. A.

Hong, A.

M. Ingaramo, A. G. York, P. Wawrzusin, O. Milberg, A. Hong, R. Weigert, H. Shroff, and G. H. Patterson, Proc. Natl. Acad. Sci. USA 111, 5254 (2014).
[Crossref]

Ingaramo, M.

M. Ingaramo, A. G. York, P. Wawrzusin, O. Milberg, A. Hong, R. Weigert, H. Shroff, and G. H. Patterson, Proc. Natl. Acad. Sci. USA 111, 5254 (2014).
[Crossref]

Kasai, A.

K. Watanabe, A. Palonpon, N. Smith, L. Chiu, A. Kasai, H. Hashimoto, S. Kawata, and K. Fujita, Nat. Commun. 6, 10095 (2015).
[Crossref]

Kawata, S.

K. Watanabe, A. Palonpon, N. Smith, L. Chiu, A. Kasai, H. Hashimoto, S. Kawata, and K. Fujita, Nat. Commun. 6, 10095 (2015).
[Crossref]

Kehlenbach, R. H.

O. Schulz, C. Pieper, M. Clever, J. Pfaff, A. Ruhlandt, R. H. Kehlenbach, F. S. Wouters, J. Großhans, G. Bunt, and J. Enderlein, Proc. Natl. Acad. Sci. USA 110, 21000 (2013).
[Crossref]

Kudenov, M. W.

N. Hagen and M. W. Kudenov, Opt. Eng. 52, 090901 (2013).
[Crossref]

Manders, E. M. M.

Milberg, O.

M. Ingaramo, A. G. York, P. Wawrzusin, O. Milberg, A. Hong, R. Weigert, H. Shroff, and G. H. Patterson, Proc. Natl. Acad. Sci. USA 111, 5254 (2014).
[Crossref]

Mione, M.

A. G. York, S. H. Parekh, D. Dalle Nogare, R. S. Fischer, K. Temprine, M. Mione, A. B. Chitnis, C. A. Combs, and H. Shroff, Nat. Methods 9, 749 (2012).
[Crossref]

Müller, C.

C. Müller and J. Enderlein, Phys. Rev. Lett. 104, 198101 (2010).
[Crossref]

Nahidi Azar, L.

Palonpon, A.

K. Watanabe, A. Palonpon, N. Smith, L. Chiu, A. Kasai, H. Hashimoto, S. Kawata, and K. Fujita, Nat. Commun. 6, 10095 (2015).
[Crossref]

Parekh, S. H.

A. G. York, S. H. Parekh, D. Dalle Nogare, R. S. Fischer, K. Temprine, M. Mione, A. B. Chitnis, C. A. Combs, and H. Shroff, Nat. Methods 9, 749 (2012).
[Crossref]

Patterson, G. H.

M. Ingaramo, A. G. York, P. Wawrzusin, O. Milberg, A. Hong, R. Weigert, H. Shroff, and G. H. Patterson, Proc. Natl. Acad. Sci. USA 111, 5254 (2014).
[Crossref]

Pfaff, J.

O. Schulz, C. Pieper, M. Clever, J. Pfaff, A. Ruhlandt, R. H. Kehlenbach, F. S. Wouters, J. Großhans, G. Bunt, and J. Enderlein, Proc. Natl. Acad. Sci. USA 110, 21000 (2013).
[Crossref]

Pieper, C.

O. Schulz, C. Pieper, M. Clever, J. Pfaff, A. Ruhlandt, R. H. Kehlenbach, F. S. Wouters, J. Großhans, G. Bunt, and J. Enderlein, Proc. Natl. Acad. Sci. USA 110, 21000 (2013).
[Crossref]

Roth, S.

S. Roth, C. J. R. Sheppard, and R. Heintzmann, Opt. Lett. 41, 2109 (2016).
[Crossref]

S. Roth, C. J. R. Sheppard, K. Wicker, and R. Heintzmann, Opt. Nanosc. 2, 5 (2013).
[Crossref]

Ruhlandt, A.

O. Schulz, C. Pieper, M. Clever, J. Pfaff, A. Ruhlandt, R. H. Kehlenbach, F. S. Wouters, J. Großhans, G. Bunt, and J. Enderlein, Proc. Natl. Acad. Sci. USA 110, 21000 (2013).
[Crossref]

Schulz, O.

O. Schulz, C. Pieper, M. Clever, J. Pfaff, A. Ruhlandt, R. H. Kehlenbach, F. S. Wouters, J. Großhans, G. Bunt, and J. Enderlein, Proc. Natl. Acad. Sci. USA 110, 21000 (2013).
[Crossref]

Sheppard, C. J. R.

S. Roth, C. J. R. Sheppard, and R. Heintzmann, Opt. Lett. 41, 2109 (2016).
[Crossref]

S. Roth, C. J. R. Sheppard, K. Wicker, and R. Heintzmann, Opt. Nanosc. 2, 5 (2013).
[Crossref]

C. J. R. Sheppard, Optik 80, 0030 (1988).

Shroff, H.

M. Ingaramo, A. G. York, P. Wawrzusin, O. Milberg, A. Hong, R. Weigert, H. Shroff, and G. H. Patterson, Proc. Natl. Acad. Sci. USA 111, 5254 (2014).
[Crossref]

A. G. York, P. Chandris, D. Dalle Nogare, J. Head, P. Wawrzusin, R. S. Fischer, A. Chitnis, and H. Shroff, Nat. Methods 10, 1122 (2013).
[Crossref]

A. G. York, S. H. Parekh, D. Dalle Nogare, R. S. Fischer, K. Temprine, M. Mione, A. B. Chitnis, C. A. Combs, and H. Shroff, Nat. Methods 9, 749 (2012).
[Crossref]

Smith, N.

K. Watanabe, A. Palonpon, N. Smith, L. Chiu, A. Kasai, H. Hashimoto, S. Kawata, and K. Fujita, Nat. Commun. 6, 10095 (2015).
[Crossref]

Stallinga, S.

Temprine, K.

A. G. York, S. H. Parekh, D. Dalle Nogare, R. S. Fischer, K. Temprine, M. Mione, A. B. Chitnis, C. A. Combs, and H. Shroff, Nat. Methods 9, 749 (2012).
[Crossref]

Timmermans, W.

Watanabe, K.

K. Watanabe, A. Palonpon, N. Smith, L. Chiu, A. Kasai, H. Hashimoto, S. Kawata, and K. Fujita, Nat. Commun. 6, 10095 (2015).
[Crossref]

Wawrzusin, P.

M. Ingaramo, A. G. York, P. Wawrzusin, O. Milberg, A. Hong, R. Weigert, H. Shroff, and G. H. Patterson, Proc. Natl. Acad. Sci. USA 111, 5254 (2014).
[Crossref]

A. G. York, P. Chandris, D. Dalle Nogare, J. Head, P. Wawrzusin, R. S. Fischer, A. Chitnis, and H. Shroff, Nat. Methods 10, 1122 (2013).
[Crossref]

Weigert, R.

M. Ingaramo, A. G. York, P. Wawrzusin, O. Milberg, A. Hong, R. Weigert, H. Shroff, and G. H. Patterson, Proc. Natl. Acad. Sci. USA 111, 5254 (2014).
[Crossref]

Wicker, K.

S. Roth, C. J. R. Sheppard, K. Wicker, and R. Heintzmann, Opt. Nanosc. 2, 5 (2013).
[Crossref]

Wouters, F. S.

O. Schulz, C. Pieper, M. Clever, J. Pfaff, A. Ruhlandt, R. H. Kehlenbach, F. S. Wouters, J. Großhans, G. Bunt, and J. Enderlein, Proc. Natl. Acad. Sci. USA 110, 21000 (2013).
[Crossref]

York, A. G.

M. Ingaramo, A. G. York, P. Wawrzusin, O. Milberg, A. Hong, R. Weigert, H. Shroff, and G. H. Patterson, Proc. Natl. Acad. Sci. USA 111, 5254 (2014).
[Crossref]

A. G. York, P. Chandris, D. Dalle Nogare, J. Head, P. Wawrzusin, R. S. Fischer, A. Chitnis, and H. Shroff, Nat. Methods 10, 1122 (2013).
[Crossref]

A. G. York, S. H. Parekh, D. Dalle Nogare, R. S. Fischer, K. Temprine, M. Mione, A. B. Chitnis, C. A. Combs, and H. Shroff, Nat. Methods 9, 749 (2012).
[Crossref]

Zeelenberg, C. H. C.

Biomed. Opt. Express (1)

Nat. Commun. (1)

K. Watanabe, A. Palonpon, N. Smith, L. Chiu, A. Kasai, H. Hashimoto, S. Kawata, and K. Fujita, Nat. Commun. 6, 10095 (2015).
[Crossref]

Nat. Methods (2)

A. G. York, S. H. Parekh, D. Dalle Nogare, R. S. Fischer, K. Temprine, M. Mione, A. B. Chitnis, C. A. Combs, and H. Shroff, Nat. Methods 9, 749 (2012).
[Crossref]

A. G. York, P. Chandris, D. Dalle Nogare, J. Head, P. Wawrzusin, R. S. Fischer, A. Chitnis, and H. Shroff, Nat. Methods 10, 1122 (2013).
[Crossref]

Opt. Eng. (1)

N. Hagen and M. W. Kudenov, Opt. Eng. 52, 090901 (2013).
[Crossref]

Opt. Lett. (1)

Opt. Nanosc. (1)

S. Roth, C. J. R. Sheppard, K. Wicker, and R. Heintzmann, Opt. Nanosc. 2, 5 (2013).
[Crossref]

Optik (1)

C. J. R. Sheppard, Optik 80, 0030 (1988).

Phys. Chem. Chem. Phys. (1)

T. Deckert-Gaudig and V. Deckert, Phys. Chem. Chem. Phys. 12, 12040 (2010).
[Crossref]

Phys. Rev. Lett. (1)

C. Müller and J. Enderlein, Phys. Rev. Lett. 104, 198101 (2010).
[Crossref]

Proc. Natl. Acad. Sci. USA (2)

O. Schulz, C. Pieper, M. Clever, J. Pfaff, A. Ruhlandt, R. H. Kehlenbach, F. S. Wouters, J. Großhans, G. Bunt, and J. Enderlein, Proc. Natl. Acad. Sci. USA 110, 21000 (2013).
[Crossref]

M. Ingaramo, A. G. York, P. Wawrzusin, O. Milberg, A. Hong, R. Weigert, H. Shroff, and G. H. Patterson, Proc. Natl. Acad. Sci. USA 111, 5254 (2014).
[Crossref]

Other (1)

M. Lamy de la Chapelle, P. Gucciardi, and N. Lidgi-Guigui, eds., Handbook of Enhanced Spectroscopy (Pan Stanford Publishing Pte Ltd., 2015).

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Figures (5)

Fig. 1.
Fig. 1. (a) Standard fiber-coupled confocal Raman microscope setup. (b) Our suggested modification, which consists of a hexagonal lenslet array and a matching round-to-linear fiber bundle. The linear end is plugged into the spectrometer and facilitates the collection of seven spectra in parallel. An additional magnification adjusts the system to its optimal performance. A closeup of the lenslet array is given in (c).
Fig. 2.
Fig. 2. (a) Camera frame at one scan point showing the seven acquired spectra is depicted. (b) Spectra at the same point in space with and without the pixel reassignment are shown. The lower panel (red curve) corresponds to the difference of the two spectra.
Fig. 3.
Fig. 3. Simulation results showing the ideal pitch of the lenslet array in units of Airy disks to achieve the highest resolution gain, i.e., improvement in resolution compared to a standard, single-fiber-based microscope with the same light collection efficiency (red, solid curve) and the associated intensity gain compared to a single fiber yielding the same resolution (green, dashed curve); the inset shows a simulated image of the PSF at the position of the lenslet array.
Fig. 4.
Fig. 4. Confocal images constructed from the data collected by each individual fiber. Lateral shifts according to the fiber position have to be compensated for before summing the signals to achieve the resolution gain associated with ISM.
Fig. 5.
Fig. 5. Measurements on a PS/PMMA mixture; first column, confocal Raman images without the pixel reassignment; second column, with the pixel reassignment. To highlight the resolution gain, the signal profiles through the bulk of the PS beads are shown in (b).

Equations (1)

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h tot n = ( h det * L n ) · h ex .

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