Abstract

We present a method for obtaining a position-dependent absorption coefficient from near-field scanning optical transmission microscopy. We show that the optical transmission intensity can be combined with the topography, resulting into an absorption coefficient that simplifies the analysis of different materials within a sample. The method is tested with the dye rhodamine 6G, and we show some analysis in biological samples such as bacteria Klebsiella pneumoniae and Pseudomonas aeruginosa. The calculated absorption coefficient images show important details of the bacteria, in particular for P. aeruginosa, in which membrane vesicles are clearly seen.

© 2003 Optical Society of America

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  1. D. W. Pohl, W. Denk, M. Lanz, “Optical stethoscopy: image recording with resolution λ/20,” Appl. Phys. Lett. 44, 651–653 (1984).
    [CrossRef]
  2. U. Dürig, D. W. Pohl, F. Rohner, “Near-field optical-scanning microscopy,” J. Appl. Phys. 59, 3318–3327 (1986).
    [CrossRef]
  3. E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, R. L. Kostelak, “Breaking the diffraction barrier: optical microscopy on a nanometric scale,” Science 251, 1468–1470 (1991).
    [CrossRef] [PubMed]
  4. E. Betzig, P. L. Finn, J. S. Weiner, “Combined shear force and near-field scanning optical microscopy,” Appl. Phys. Lett. 60, 2484–2486 (1992).
    [CrossRef]
  5. T. Enderle, T. Ha, D. S. Chemla, S. Weiss, “Near-field fluorescence microscopy of cells,” Ultramicroscopy 71, 303–309 (1998).
    [CrossRef] [PubMed]
  6. A. Penzkofer, E. Drotleff, W. Holzer, “Optical constants measurement of single-layer thin films on transparent substrates,” Opt. Commun. 158, 221–230 (1998).
    [CrossRef]
  7. R. Carminati, A. Madrazo, M. Nieto-Vesperinas, J.-J. Greffet, “Optical content and resolution of near-field optical images: influence of the operating mode,” J. Appl. Phys. 82, 501–509 (1997).
    [CrossRef]
  8. G. W. Bryant, “Probing quantum nanostructures with near-field optical microscopy and vice versa,” Appl. Phys. Lett. 72, 768–770 (1998).
    [CrossRef]
  9. R. Stevenson, D. Richards, “The use of a near-field probe for the study of semiconductor heterostructures,” Semicond. Sci. Technol. 13, 882–886 (1998).
    [CrossRef]
  10. H. B. Silva, J. A. Toledo, K. Silveira Filho, G. Weber, A. M. de Paula, “Near-field scanning optical transmission images in Pseudomonas aeruginosa,” Acta Microscopica 10, 99–102 (2001).
  11. N. Ben-Ami, A. Radko, U. Ben-Ami, K. Lieberman, Z. Rothman, I. Rabin, A. Lewis, “Near-field optical imaging of unstained bacteria: comparison with normal atomic force and far-field optical microscopy in air and aqueous media,” Ultramicroscopy 71, 321–325 (1998).
    [CrossRef] [PubMed]
  12. J. L. Kadurugamuwa, T. J. Beveridge, “Virulence factors are released from Pseudomonas aeruginosa in association with membrane vesicles during normal growth and exposure to gentamicin: a novel mechanism of enzyme secretion,” J. Bacteriol. 177, 3998–4008 (1995).
    [PubMed]

2001 (1)

H. B. Silva, J. A. Toledo, K. Silveira Filho, G. Weber, A. M. de Paula, “Near-field scanning optical transmission images in Pseudomonas aeruginosa,” Acta Microscopica 10, 99–102 (2001).

1998 (5)

N. Ben-Ami, A. Radko, U. Ben-Ami, K. Lieberman, Z. Rothman, I. Rabin, A. Lewis, “Near-field optical imaging of unstained bacteria: comparison with normal atomic force and far-field optical microscopy in air and aqueous media,” Ultramicroscopy 71, 321–325 (1998).
[CrossRef] [PubMed]

T. Enderle, T. Ha, D. S. Chemla, S. Weiss, “Near-field fluorescence microscopy of cells,” Ultramicroscopy 71, 303–309 (1998).
[CrossRef] [PubMed]

A. Penzkofer, E. Drotleff, W. Holzer, “Optical constants measurement of single-layer thin films on transparent substrates,” Opt. Commun. 158, 221–230 (1998).
[CrossRef]

G. W. Bryant, “Probing quantum nanostructures with near-field optical microscopy and vice versa,” Appl. Phys. Lett. 72, 768–770 (1998).
[CrossRef]

R. Stevenson, D. Richards, “The use of a near-field probe for the study of semiconductor heterostructures,” Semicond. Sci. Technol. 13, 882–886 (1998).
[CrossRef]

1997 (1)

R. Carminati, A. Madrazo, M. Nieto-Vesperinas, J.-J. Greffet, “Optical content and resolution of near-field optical images: influence of the operating mode,” J. Appl. Phys. 82, 501–509 (1997).
[CrossRef]

1995 (1)

J. L. Kadurugamuwa, T. J. Beveridge, “Virulence factors are released from Pseudomonas aeruginosa in association with membrane vesicles during normal growth and exposure to gentamicin: a novel mechanism of enzyme secretion,” J. Bacteriol. 177, 3998–4008 (1995).
[PubMed]

1992 (1)

E. Betzig, P. L. Finn, J. S. Weiner, “Combined shear force and near-field scanning optical microscopy,” Appl. Phys. Lett. 60, 2484–2486 (1992).
[CrossRef]

1991 (1)

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, R. L. Kostelak, “Breaking the diffraction barrier: optical microscopy on a nanometric scale,” Science 251, 1468–1470 (1991).
[CrossRef] [PubMed]

1986 (1)

U. Dürig, D. W. Pohl, F. Rohner, “Near-field optical-scanning microscopy,” J. Appl. Phys. 59, 3318–3327 (1986).
[CrossRef]

1984 (1)

D. W. Pohl, W. Denk, M. Lanz, “Optical stethoscopy: image recording with resolution λ/20,” Appl. Phys. Lett. 44, 651–653 (1984).
[CrossRef]

Ben-Ami, N.

N. Ben-Ami, A. Radko, U. Ben-Ami, K. Lieberman, Z. Rothman, I. Rabin, A. Lewis, “Near-field optical imaging of unstained bacteria: comparison with normal atomic force and far-field optical microscopy in air and aqueous media,” Ultramicroscopy 71, 321–325 (1998).
[CrossRef] [PubMed]

Ben-Ami, U.

N. Ben-Ami, A. Radko, U. Ben-Ami, K. Lieberman, Z. Rothman, I. Rabin, A. Lewis, “Near-field optical imaging of unstained bacteria: comparison with normal atomic force and far-field optical microscopy in air and aqueous media,” Ultramicroscopy 71, 321–325 (1998).
[CrossRef] [PubMed]

Betzig, E.

E. Betzig, P. L. Finn, J. S. Weiner, “Combined shear force and near-field scanning optical microscopy,” Appl. Phys. Lett. 60, 2484–2486 (1992).
[CrossRef]

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, R. L. Kostelak, “Breaking the diffraction barrier: optical microscopy on a nanometric scale,” Science 251, 1468–1470 (1991).
[CrossRef] [PubMed]

Beveridge, T. J.

J. L. Kadurugamuwa, T. J. Beveridge, “Virulence factors are released from Pseudomonas aeruginosa in association with membrane vesicles during normal growth and exposure to gentamicin: a novel mechanism of enzyme secretion,” J. Bacteriol. 177, 3998–4008 (1995).
[PubMed]

Bryant, G. W.

G. W. Bryant, “Probing quantum nanostructures with near-field optical microscopy and vice versa,” Appl. Phys. Lett. 72, 768–770 (1998).
[CrossRef]

Carminati, R.

R. Carminati, A. Madrazo, M. Nieto-Vesperinas, J.-J. Greffet, “Optical content and resolution of near-field optical images: influence of the operating mode,” J. Appl. Phys. 82, 501–509 (1997).
[CrossRef]

Chemla, D. S.

T. Enderle, T. Ha, D. S. Chemla, S. Weiss, “Near-field fluorescence microscopy of cells,” Ultramicroscopy 71, 303–309 (1998).
[CrossRef] [PubMed]

de Paula, A. M.

H. B. Silva, J. A. Toledo, K. Silveira Filho, G. Weber, A. M. de Paula, “Near-field scanning optical transmission images in Pseudomonas aeruginosa,” Acta Microscopica 10, 99–102 (2001).

Denk, W.

D. W. Pohl, W. Denk, M. Lanz, “Optical stethoscopy: image recording with resolution λ/20,” Appl. Phys. Lett. 44, 651–653 (1984).
[CrossRef]

Drotleff, E.

A. Penzkofer, E. Drotleff, W. Holzer, “Optical constants measurement of single-layer thin films on transparent substrates,” Opt. Commun. 158, 221–230 (1998).
[CrossRef]

Dürig, U.

U. Dürig, D. W. Pohl, F. Rohner, “Near-field optical-scanning microscopy,” J. Appl. Phys. 59, 3318–3327 (1986).
[CrossRef]

Enderle, T.

T. Enderle, T. Ha, D. S. Chemla, S. Weiss, “Near-field fluorescence microscopy of cells,” Ultramicroscopy 71, 303–309 (1998).
[CrossRef] [PubMed]

Finn, P. L.

E. Betzig, P. L. Finn, J. S. Weiner, “Combined shear force and near-field scanning optical microscopy,” Appl. Phys. Lett. 60, 2484–2486 (1992).
[CrossRef]

Greffet, J.-J.

R. Carminati, A. Madrazo, M. Nieto-Vesperinas, J.-J. Greffet, “Optical content and resolution of near-field optical images: influence of the operating mode,” J. Appl. Phys. 82, 501–509 (1997).
[CrossRef]

Ha, T.

T. Enderle, T. Ha, D. S. Chemla, S. Weiss, “Near-field fluorescence microscopy of cells,” Ultramicroscopy 71, 303–309 (1998).
[CrossRef] [PubMed]

Harris, T. D.

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, R. L. Kostelak, “Breaking the diffraction barrier: optical microscopy on a nanometric scale,” Science 251, 1468–1470 (1991).
[CrossRef] [PubMed]

Holzer, W.

A. Penzkofer, E. Drotleff, W. Holzer, “Optical constants measurement of single-layer thin films on transparent substrates,” Opt. Commun. 158, 221–230 (1998).
[CrossRef]

Kadurugamuwa, J. L.

J. L. Kadurugamuwa, T. J. Beveridge, “Virulence factors are released from Pseudomonas aeruginosa in association with membrane vesicles during normal growth and exposure to gentamicin: a novel mechanism of enzyme secretion,” J. Bacteriol. 177, 3998–4008 (1995).
[PubMed]

Kostelak, R. L.

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, R. L. Kostelak, “Breaking the diffraction barrier: optical microscopy on a nanometric scale,” Science 251, 1468–1470 (1991).
[CrossRef] [PubMed]

Lanz, M.

D. W. Pohl, W. Denk, M. Lanz, “Optical stethoscopy: image recording with resolution λ/20,” Appl. Phys. Lett. 44, 651–653 (1984).
[CrossRef]

Lewis, A.

N. Ben-Ami, A. Radko, U. Ben-Ami, K. Lieberman, Z. Rothman, I. Rabin, A. Lewis, “Near-field optical imaging of unstained bacteria: comparison with normal atomic force and far-field optical microscopy in air and aqueous media,” Ultramicroscopy 71, 321–325 (1998).
[CrossRef] [PubMed]

Lieberman, K.

N. Ben-Ami, A. Radko, U. Ben-Ami, K. Lieberman, Z. Rothman, I. Rabin, A. Lewis, “Near-field optical imaging of unstained bacteria: comparison with normal atomic force and far-field optical microscopy in air and aqueous media,” Ultramicroscopy 71, 321–325 (1998).
[CrossRef] [PubMed]

Madrazo, A.

R. Carminati, A. Madrazo, M. Nieto-Vesperinas, J.-J. Greffet, “Optical content and resolution of near-field optical images: influence of the operating mode,” J. Appl. Phys. 82, 501–509 (1997).
[CrossRef]

Nieto-Vesperinas, M.

R. Carminati, A. Madrazo, M. Nieto-Vesperinas, J.-J. Greffet, “Optical content and resolution of near-field optical images: influence of the operating mode,” J. Appl. Phys. 82, 501–509 (1997).
[CrossRef]

Penzkofer, A.

A. Penzkofer, E. Drotleff, W. Holzer, “Optical constants measurement of single-layer thin films on transparent substrates,” Opt. Commun. 158, 221–230 (1998).
[CrossRef]

Pohl, D. W.

U. Dürig, D. W. Pohl, F. Rohner, “Near-field optical-scanning microscopy,” J. Appl. Phys. 59, 3318–3327 (1986).
[CrossRef]

D. W. Pohl, W. Denk, M. Lanz, “Optical stethoscopy: image recording with resolution λ/20,” Appl. Phys. Lett. 44, 651–653 (1984).
[CrossRef]

Rabin, I.

N. Ben-Ami, A. Radko, U. Ben-Ami, K. Lieberman, Z. Rothman, I. Rabin, A. Lewis, “Near-field optical imaging of unstained bacteria: comparison with normal atomic force and far-field optical microscopy in air and aqueous media,” Ultramicroscopy 71, 321–325 (1998).
[CrossRef] [PubMed]

Radko, A.

N. Ben-Ami, A. Radko, U. Ben-Ami, K. Lieberman, Z. Rothman, I. Rabin, A. Lewis, “Near-field optical imaging of unstained bacteria: comparison with normal atomic force and far-field optical microscopy in air and aqueous media,” Ultramicroscopy 71, 321–325 (1998).
[CrossRef] [PubMed]

Richards, D.

R. Stevenson, D. Richards, “The use of a near-field probe for the study of semiconductor heterostructures,” Semicond. Sci. Technol. 13, 882–886 (1998).
[CrossRef]

Rohner, F.

U. Dürig, D. W. Pohl, F. Rohner, “Near-field optical-scanning microscopy,” J. Appl. Phys. 59, 3318–3327 (1986).
[CrossRef]

Rothman, Z.

N. Ben-Ami, A. Radko, U. Ben-Ami, K. Lieberman, Z. Rothman, I. Rabin, A. Lewis, “Near-field optical imaging of unstained bacteria: comparison with normal atomic force and far-field optical microscopy in air and aqueous media,” Ultramicroscopy 71, 321–325 (1998).
[CrossRef] [PubMed]

Silva, H. B.

H. B. Silva, J. A. Toledo, K. Silveira Filho, G. Weber, A. M. de Paula, “Near-field scanning optical transmission images in Pseudomonas aeruginosa,” Acta Microscopica 10, 99–102 (2001).

Silveira Filho, K.

H. B. Silva, J. A. Toledo, K. Silveira Filho, G. Weber, A. M. de Paula, “Near-field scanning optical transmission images in Pseudomonas aeruginosa,” Acta Microscopica 10, 99–102 (2001).

Stevenson, R.

R. Stevenson, D. Richards, “The use of a near-field probe for the study of semiconductor heterostructures,” Semicond. Sci. Technol. 13, 882–886 (1998).
[CrossRef]

Toledo, J. A.

H. B. Silva, J. A. Toledo, K. Silveira Filho, G. Weber, A. M. de Paula, “Near-field scanning optical transmission images in Pseudomonas aeruginosa,” Acta Microscopica 10, 99–102 (2001).

Trautman, J. K.

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, R. L. Kostelak, “Breaking the diffraction barrier: optical microscopy on a nanometric scale,” Science 251, 1468–1470 (1991).
[CrossRef] [PubMed]

Weber, G.

H. B. Silva, J. A. Toledo, K. Silveira Filho, G. Weber, A. M. de Paula, “Near-field scanning optical transmission images in Pseudomonas aeruginosa,” Acta Microscopica 10, 99–102 (2001).

Weiner, J. S.

E. Betzig, P. L. Finn, J. S. Weiner, “Combined shear force and near-field scanning optical microscopy,” Appl. Phys. Lett. 60, 2484–2486 (1992).
[CrossRef]

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, R. L. Kostelak, “Breaking the diffraction barrier: optical microscopy on a nanometric scale,” Science 251, 1468–1470 (1991).
[CrossRef] [PubMed]

Weiss, S.

T. Enderle, T. Ha, D. S. Chemla, S. Weiss, “Near-field fluorescence microscopy of cells,” Ultramicroscopy 71, 303–309 (1998).
[CrossRef] [PubMed]

Acta Microscopica (1)

H. B. Silva, J. A. Toledo, K. Silveira Filho, G. Weber, A. M. de Paula, “Near-field scanning optical transmission images in Pseudomonas aeruginosa,” Acta Microscopica 10, 99–102 (2001).

Appl. Phys. Lett. (3)

D. W. Pohl, W. Denk, M. Lanz, “Optical stethoscopy: image recording with resolution λ/20,” Appl. Phys. Lett. 44, 651–653 (1984).
[CrossRef]

E. Betzig, P. L. Finn, J. S. Weiner, “Combined shear force and near-field scanning optical microscopy,” Appl. Phys. Lett. 60, 2484–2486 (1992).
[CrossRef]

G. W. Bryant, “Probing quantum nanostructures with near-field optical microscopy and vice versa,” Appl. Phys. Lett. 72, 768–770 (1998).
[CrossRef]

J. Appl. Phys. (2)

R. Carminati, A. Madrazo, M. Nieto-Vesperinas, J.-J. Greffet, “Optical content and resolution of near-field optical images: influence of the operating mode,” J. Appl. Phys. 82, 501–509 (1997).
[CrossRef]

U. Dürig, D. W. Pohl, F. Rohner, “Near-field optical-scanning microscopy,” J. Appl. Phys. 59, 3318–3327 (1986).
[CrossRef]

J. Bacteriol. (1)

J. L. Kadurugamuwa, T. J. Beveridge, “Virulence factors are released from Pseudomonas aeruginosa in association with membrane vesicles during normal growth and exposure to gentamicin: a novel mechanism of enzyme secretion,” J. Bacteriol. 177, 3998–4008 (1995).
[PubMed]

Opt. Commun. (1)

A. Penzkofer, E. Drotleff, W. Holzer, “Optical constants measurement of single-layer thin films on transparent substrates,” Opt. Commun. 158, 221–230 (1998).
[CrossRef]

Science (1)

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, R. L. Kostelak, “Breaking the diffraction barrier: optical microscopy on a nanometric scale,” Science 251, 1468–1470 (1991).
[CrossRef] [PubMed]

Semicond. Sci. Technol. (1)

R. Stevenson, D. Richards, “The use of a near-field probe for the study of semiconductor heterostructures,” Semicond. Sci. Technol. 13, 882–886 (1998).
[CrossRef]

Ultramicroscopy (2)

T. Enderle, T. Ha, D. S. Chemla, S. Weiss, “Near-field fluorescence microscopy of cells,” Ultramicroscopy 71, 303–309 (1998).
[CrossRef] [PubMed]

N. Ben-Ami, A. Radko, U. Ben-Ami, K. Lieberman, Z. Rothman, I. Rabin, A. Lewis, “Near-field optical imaging of unstained bacteria: comparison with normal atomic force and far-field optical microscopy in air and aqueous media,” Ultramicroscopy 71, 321–325 (1998).
[CrossRef] [PubMed]

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Figures (3)

Fig. 1
Fig. 1

Near-field images for rhodamine 6G: topography (top), optical transmission (center), and calculated absorption coefficient (bottom). The curves indicate the profiles along the indicated lines.

Fig. 2
Fig. 2

Near-field images for K. pneumoniae: topography (top), optical transmission (center), and calculated absorption coefficient (bottom). The curves indicate the profiles along the indicated lines.

Fig. 3
Fig. 3

Near-field images for P. aeruginosa: topography (top), optical transmission (center), and calculated absorption coefficient (bottom). The curves indicate the profiles along the indicated lines.

Equations (2)

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Iz=I0 exp-αz,
αx, y=- lnIx, y, z/I0Tx, y,

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