Fast determination of saturation intensity and maximum emission rate by single-emitter imaging

doi:10.1364/OE.14.009350

Fast determination of saturation intensity and maximum emission rate by single-emitter imaging

J. Y. P. Butter and B. Hecht »View Author Affiliations

Optics Express, Vol. 14, Issue 20, pp. 9350-9357 (2006)
http://dx.doi.org/10.1364/OE.14.009350

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Abstract

We investigate the dependence of the spot size in single-emitter confocal imaging on the degree of saturation. We show that single-emitter spots are broadened and flattened significantly already at excitation intensities well below saturation. The resulting single-emitter spot shapes thus deviate significantly from the excitation point spread function. We show and support by Monte Carlo simulations that fitting of a single spot is sufficient to extract the saturation intensity and the maximum emission rate of a single emitter with high accuracy. Our results will be of interest in all areas of single-emitter studies.

OCIS Codes
(180.1790) Microscopy : Confocal microscopy
(260.2510) Physical optics : Fluorescence
(270.0270) Quantum optics : Quantum optics
(300.6280) Spectroscopy : Spectroscopy, fluorescence and luminescence

ToC Category:
Microscopy

History
Original Manuscript: July 25, 2006
Revised Manuscript: September 11, 2006
Manuscript Accepted: September 11, 2006
Published: October 2, 2006

Virtual Issues
Vol. 1, Iss. 11 Virtual Journal for Biomedical Optics

Citation
J. Y. P. Butter and B. Hecht, "Fast determination of saturation intensity and maximum emission rate by single-emitter imaging," Opt. Express 14, 9350-9357 (2006)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-14-20-9350

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References

T. Basché, W. E. Moerner, M. Orrit, and U. P. Wild, eds., Single-Molecule Optical Detection, Imaging and Spectroscopy (VCH Verlagsgesellschaft, Weinheim, 1997).
W. P. Ambrose, T. Basché, and W. E. Moerner, "Detection and spectroscopy of single pentacene molecules in a p-terphenyl crystal by means of fluorescence excitation," J. Chem. Phys. 95, 7150-7163 (1991). [CrossRef]
L. Novotny and B. Hecht, Principles of Nano-Optics (Cambridge University Press, 2006).
N. Bobroff, "Position Measurement with a Resolution and Noise-Limited Instrument," Rev. Sci. Instrum. 57, 1152-1157 (1986). [CrossRef]
T. Schmidt, G. J. Schütz,W. Baumgartner, H. J. Gruber, and H. Schindler, "Imaging of single molecule diffusion," Proc. Natl. Acad. Sci. USA 93, 2926-2929 (1996). [CrossRef] [PubMed]
A. Bloeß, Y. Durand, M. Matsushita, H. van der Meer, G. J. Brakenhoff, and J. Schmidt, "Optical far-field microscopy of single molecules with 3.4 nm lateral resolution," J. Microsc. 205, 76-85 (2002). [CrossRef] [PubMed]
A. M. van Oijen, J. K¨ohler, J. Schmidt, M. Müller, and G. J. Brakenhoff, "3-Dimensional super-resolution by spectrally selective imaging," Chem. Phys. Lett. 292, 183-187 (1998). [CrossRef]
A. Yildiz, J. N. Forkey, S. A. McKinney, T. Ha, Y. E. Goldman, and P. R. Selvin, "Myosin V Walks Hand-Over- Hand: Single Fluorophore Imaging with 1.5-nm Localization," Science 300, 2061-2065 (2003). [CrossRef] [PubMed]
J.-M. Segura, A. Renn, and B. Hecht, "A sample-scanning confocal optical microscope for cryogenic operation," Rev. Sci. Instrum. 71, 1706-1711 (2000). [CrossRef]
S. Kummer, F. Kulzer, R. Kettner, T. Basché, C. Tietz, C. Glowatz, and C. Kryschi, "Absorption, excitation and emission spectroscopy of terrylene in p-terphenyl: Bulk measurements and single molecule studies," J. Chem. Phys. 107, 7673-7684 (1997). [CrossRef]
S. Kummer, T. Basché, and C. Bräuchle, "Terrylene in p-terphenyl: a novel single crystalline system for single molecule spectroscopy at low temperatures," Chem. Phys. Lett. 229, 309-316 (1994). [CrossRef]
P. Bordat and R. Brown, "Molecular mechanisms of photo-induced spectral diffusion of single terrylene molecules in p-terphenyl," J. Chem. Phys. 116, 229-236 (2002). [CrossRef]
J. Y. P. Butter and B. Hecht, "Aperture scanning near-field optical microscopy and spectroscopy of single terrylene molecules at 1.8 K," Nanotechnology 17, 1547-1550 (2006). [CrossRef]
T. Plakhotnik, W. E. Moerner, V. Palm, and U. P. Wild, "Single molecule spectroscopy: maximum emission rate and saturation intensity," Opt. Commun. 114, 83-88 (1995). [CrossRef]
J. Bernard, L. Fleury, H. Talon, and M. Orrit, "Photon bunching in the fluorescence from single molecules: A probe for intersystem crossing," J. Chem. Phys. 98, 850-859 (1993). [CrossRef]
H. de Vries and D. A. Wiersma, "Photophysical and photochemical molecular hole burning theory," J. Chem. Phys. 72, 1851-1863 (1980). [CrossRef]
P. Brémaud, An introdcution to probablistic modeling (Springer-Verlag, 1994).
P. R. Bevington and D. K. Robinson, Data reduction and error analysis for the physical sciences, 2nd ed. (WCB/McGraw-Hill, 1992).

Ambrose, W. P.
Basché, T.
Baumgartner, W.
Bernard, J.
Bloeß, A.
Bobroff, N.
Bordat, P.
Brakenhoff, G. J.
Bräuchle, C.
Brown, R.
Butter, J. Y. P.
de Vries, H.
Durand, Y.
Fleury, L.
Forkey, J. N.
Glowatz, C.
Goldman, Y. E.
Gruber, H. J.
Ha, T.
Hecht, B.
Kettner, R.
Kryschi, C.
Kulzer, F.
Kummer, S.
Matsushita, M.
McKinney, S. A.
Moerner, W. E.
Orrit, M.
Palm, V.
Plakhotnik, T.
Renn, A.
Schindler, H.
Schmidt, J.
Schmidt, T.
Schütz, G. J.
Segura, J.-M.
Selvin, P. R.
Talon, H.
Tietz, C.
van der Meer, H.
van Oijen, A. M.
Wiersma, D. A.
Wild, U. P.
Yildiz, A.

Chem. Phys. Lett.

A. M. van Oijen, J. K¨ohler, J. Schmidt, M. Müller, and G. J. Brakenhoff, "3-Dimensional super-resolution by spectrally selective imaging," Chem. Phys. Lett. 292, 183-187 (1998). [CrossRef]
S. Kummer, T. Basché, and C. Bräuchle, "Terrylene in p-terphenyl: a novel single crystalline system for single molecule spectroscopy at low temperatures," Chem. Phys. Lett. 229, 309-316 (1994). [CrossRef]

J. Chem. Phys.

P. Bordat and R. Brown, "Molecular mechanisms of photo-induced spectral diffusion of single terrylene molecules in p-terphenyl," J. Chem. Phys. 116, 229-236 (2002). [CrossRef]
J. Bernard, L. Fleury, H. Talon, and M. Orrit, "Photon bunching in the fluorescence from single molecules: A probe for intersystem crossing," J. Chem. Phys. 98, 850-859 (1993). [CrossRef]
H. de Vries and D. A. Wiersma, "Photophysical and photochemical molecular hole burning theory," J. Chem. Phys. 72, 1851-1863 (1980). [CrossRef]
W. P. Ambrose, T. Basché, and W. E. Moerner, "Detection and spectroscopy of single pentacene molecules in a p-terphenyl crystal by means of fluorescence excitation," J. Chem. Phys. 95, 7150-7163 (1991). [CrossRef]
S. Kummer, F. Kulzer, R. Kettner, T. Basché, C. Tietz, C. Glowatz, and C. Kryschi, "Absorption, excitation and emission spectroscopy of terrylene in p-terphenyl: Bulk measurements and single molecule studies," J. Chem. Phys. 107, 7673-7684 (1997). [CrossRef]

J. Microsc.

A. Bloeß, Y. Durand, M. Matsushita, H. van der Meer, G. J. Brakenhoff, and J. Schmidt, "Optical far-field microscopy of single molecules with 3.4 nm lateral resolution," J. Microsc. 205, 76-85 (2002). [CrossRef] [PubMed]

Nanotechnology

J. Y. P. Butter and B. Hecht, "Aperture scanning near-field optical microscopy and spectroscopy of single terrylene molecules at 1.8 K," Nanotechnology 17, 1547-1550 (2006). [CrossRef]

Opt. Commun.

T. Plakhotnik, W. E. Moerner, V. Palm, and U. P. Wild, "Single molecule spectroscopy: maximum emission rate and saturation intensity," Opt. Commun. 114, 83-88 (1995). [CrossRef]

Proc. Natl. Acad. Sci. USA

T. Schmidt, G. J. Schütz,W. Baumgartner, H. J. Gruber, and H. Schindler, "Imaging of single molecule diffusion," Proc. Natl. Acad. Sci. USA 93, 2926-2929 (1996). [CrossRef] [PubMed]

Rev. Sci. Instrum.

N. Bobroff, "Position Measurement with a Resolution and Noise-Limited Instrument," Rev. Sci. Instrum. 57, 1152-1157 (1986). [CrossRef]
J.-M. Segura, A. Renn, and B. Hecht, "A sample-scanning confocal optical microscope for cryogenic operation," Rev. Sci. Instrum. 71, 1706-1711 (2000). [CrossRef]

Science

A. Yildiz, J. N. Forkey, S. A. McKinney, T. Ha, Y. E. Goldman, and P. R. Selvin, "Myosin V Walks Hand-Over- Hand: Single Fluorophore Imaging with 1.5-nm Localization," Science 300, 2061-2065 (2003). [CrossRef] [PubMed]

Other

T. Basché, W. E. Moerner, M. Orrit, and U. P. Wild, eds., Single-Molecule Optical Detection, Imaging and Spectroscopy (VCH Verlagsgesellschaft, Weinheim, 1997).
L. Novotny and B. Hecht, Principles of Nano-Optics (Cambridge University Press, 2006).
P. Brémaud, An introdcution to probablistic modeling (Springer-Verlag, 1994).
P. R. Bevington and D. K. Robinson, Data reduction and error analysis for the physical sciences, 2nd ed. (WCB/McGraw-Hill, 1992).

2006, Butter, Nanotechnology
2003, Yildiz, Science
2002, Bloeß, J. Microsc.
2002, Bordat, J. Chem. Phys.
2000, Segura, Rev. Sci. Instrum.
1998, van Oijen, Chem. Phys. Lett.
1997, Kummer, J. Chem. Phys.
1996, Schmidt, Proc. Natl. Acad. Sci. USA
1995, Plakhotnik, Opt. Commun.
1994, Kummer, Chem. Phys. Lett.
1993, Bernard, J. Chem. Phys.
1991, Ambrose, J. Chem. Phys.
1986, Bobroff, Rev. Sci. Instrum.
1980, de Vries, J. Chem. Phys.

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[1] T. Basché, W. E. Moerner, M. Orrit, and U. P. Wild, eds., Single-Molecule Optical Detection, Imaging and Spectroscopy (VCH Verlagsgesellschaft, Weinheim, 1997).

[2] W. P. Ambrose, T. Basché, and W. E. Moerner, "Detection and spectroscopy of single pentacene molecules in a p-terphenyl crystal by means of fluorescence excitation," J. Chem. Phys. 95, 7150-7163 (1991). [CrossRef]

[3] L. Novotny and B. Hecht, Principles of Nano-Optics (Cambridge University Press, 2006).

[4] N. Bobroff, "Position Measurement with a Resolution and Noise-Limited Instrument," Rev. Sci. Instrum. 57, 1152-1157 (1986). [CrossRef]

[5] T. Schmidt, G. J. Schütz,W. Baumgartner, H. J. Gruber, and H. Schindler, "Imaging of single molecule diffusion," Proc. Natl. Acad. Sci. USA 93, 2926-2929 (1996). [CrossRef] [PubMed]

[6] A. Bloeß, Y. Durand, M. Matsushita, H. van der Meer, G. J. Brakenhoff, and J. Schmidt, "Optical far-field microscopy of single molecules with 3.4 nm lateral resolution," J. Microsc. 205, 76-85 (2002). [CrossRef] [PubMed]

[7] A. M. van Oijen, J. K¨ohler, J. Schmidt, M. Müller, and G. J. Brakenhoff, "3-Dimensional super-resolution by spectrally selective imaging," Chem. Phys. Lett. 292, 183-187 (1998). [CrossRef]

[8] A. Yildiz, J. N. Forkey, S. A. McKinney, T. Ha, Y. E. Goldman, and P. R. Selvin, "Myosin V Walks Hand-Over- Hand: Single Fluorophore Imaging with 1.5-nm Localization," Science 300, 2061-2065 (2003). [CrossRef] [PubMed]

[9] J.-M. Segura, A. Renn, and B. Hecht, "A sample-scanning confocal optical microscope for cryogenic operation," Rev. Sci. Instrum. 71, 1706-1711 (2000). [CrossRef]

[10] S. Kummer, F. Kulzer, R. Kettner, T. Basché, C. Tietz, C. Glowatz, and C. Kryschi, "Absorption, excitation and emission spectroscopy of terrylene in p-terphenyl: Bulk measurements and single molecule studies," J. Chem. Phys. 107, 7673-7684 (1997). [CrossRef]

[11] S. Kummer, T. Basché, and C. Bräuchle, "Terrylene in p-terphenyl: a novel single crystalline system for single molecule spectroscopy at low temperatures," Chem. Phys. Lett. 229, 309-316 (1994). [CrossRef]

[12] P. Bordat and R. Brown, "Molecular mechanisms of photo-induced spectral diffusion of single terrylene molecules in p-terphenyl," J. Chem. Phys. 116, 229-236 (2002). [CrossRef]

[13] J. Y. P. Butter and B. Hecht, "Aperture scanning near-field optical microscopy and spectroscopy of single terrylene molecules at 1.8 K," Nanotechnology 17, 1547-1550 (2006). [CrossRef]

[14] T. Plakhotnik, W. E. Moerner, V. Palm, and U. P. Wild, "Single molecule spectroscopy: maximum emission rate and saturation intensity," Opt. Commun. 114, 83-88 (1995). [CrossRef]

[15] J. Bernard, L. Fleury, H. Talon, and M. Orrit, "Photon bunching in the fluorescence from single molecules: A probe for intersystem crossing," J. Chem. Phys. 98, 850-859 (1993). [CrossRef]

[16] H. de Vries and D. A. Wiersma, "Photophysical and photochemical molecular hole burning theory," J. Chem. Phys. 72, 1851-1863 (1980). [CrossRef]

[17] P. Brémaud, An introdcution to probablistic modeling (Springer-Verlag, 1994).

[18] P. R. Bevington and D. K. Robinson, Data reduction and error analysis for the physical sciences, 2nd ed. (WCB/McGraw-Hill, 1992).

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Fast determination of saturation intensity and maximum emission rate by single-emitter imaging

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Abstract

References

Chem. Phys. Lett.

J. Chem. Phys.

J. Microsc.

Nanotechnology

Opt. Commun.

Proc. Natl. Acad. Sci. USA

Rev. Sci. Instrum.

Science

Other

Cited By

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