Abstract

Fluorescence correlation spectroscopy (FCS) is an important spectroscopic technique which can be used for measuring the diffusion and thus size of fluorescing molecules at pico- to nanomolar concentrations. Recently, we introduced an extension of conventional FCS, which is called dual-focus FCS (2fFCS) and allows absolute diffusion measurements with high precision and repeatability. It was shown experimentally that the method is robust against most optical and sample artefacts which are troubling conventional FCS measurements, and is furthermore able to yield absolute values of diffusion coefficients without referencing against known standards. However, a thorough theoretical treatment of the performance of 2fFCS is still missing. The present paper aims at filling this gap. Here, we have systematically studied the performance of 2fFCS with respect to the most important optical and photophysical factors such as cover slide thick-ness, refractive index of the sample, laser beam geometry, and optical satu-ration. We show that 2fFCS has indeed a superior performance when com-pared with conventional FCS, being mostly insensitive to most potential ab-errations when working under optimized conditions.

© 2008 Optical Society of America

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    [CrossRef]
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    [CrossRef]
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    [CrossRef]
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    [CrossRef]

2008 (5)

Z. Petrasek and P. Schwille, "Precise measurement of diffusion coefficients using scanning fluorescence correlation spectroscopy," Biophys. J. 94, 1437-1448 (2008).
[CrossRef]

A. Loman, T. Dertinger, F. Koberling, and J. Enderlein, "Comparison of optical saturation effects in conventional and dual-focus fluorescence correlation spectroscopy," Chem. Phys. Lett. 459, 18-21 (2008).
[CrossRef]

C. B. Müller, A. Loman, V. Pacheco, F. Koberling, D. Willbold, W. Richtering, and J. Enderlein, "Precise Measurement of Diffusion by Multi-Color Dual-Focus Fluorescence Correlation Spectroscopy," Eur. Phys. Lett. 83, 46001 (2008).
[CrossRef]

C. B. Müller, K. Wei??, W. Richtering, A. Loman, and J. Enderlein, "Calibrating Differential Interference Contrast Microscopy with dual-focus Fluorescence Correlation Spectroscopy," Opt. Express 16, 4322-9 (2008).
[CrossRef]

M. Leutenegger and T. Lasser, "Detection efficiency in total internal reflection fluorescence microscopy," Opt. Express 16, 8519-8531 (2008).
[CrossRef]

2007 (2)

G. Donnert, C. Eggeling, and S. W Hell, "Major signal increase in fluorescence microscopy through dark-state relaxation," Nature Meth. 4, 81-86 (2007).
[CrossRef]

T. Dertinger, V. Pacheco, I. von der Hocht, R. Hartmann, I. Gregor, and J. Enderlein, "Two-focus fluorescence correlation spectroscopy: A new tool for accurate and absolute diffusion measurements," ChemPhysChem 8, 433-443 (2007).
[CrossRef]

2006 (3)

J. Humpolicková, E. Gielen, A. Benda, V. Fagulova, J. Vercammen, M. VandeVen, M. Hof, M. Ameloot, and Y. Engelborghs, "Probing Diffusion Laws within Cellular Membranes by Z-Scan Fluorescence Correlation Spectroscopy," Biophys. J. Biophys. Lett. 91, L23-L25 (2006).

J. Ries and P. Schwille, "Studying Slow Membrane Dynamics with Continuous Wave Scanning Fluorescence Correlation Spectroscopy," Biophys. J. 91, 1915-1924 (2006).
[CrossRef]

M. Leutenegger, R. Rao, R. A. Leitgeb, and T. Lasser, "Fast focus field calculations," Opt. Express 14, 11277-11291 (2006).
[CrossRef]

2005 (6)

P. R. T. Munro and P. Török, "Vectorial, high numerical aperture study of Nomarski's differential interference contrast microscope," Opt. Express 13, 6833-47 (2005).
[CrossRef]

I. Gregor and J. Enderlein "Focusing astigmatic Gaussian beams through optical systems with a high numerical aperture," Opt. Lett. 30, 2527-9 (2005).
[CrossRef]

J. Enderlein, I. Gregor, D. Patra, T. Dertinger, and U. B. Kaupp, "Performance of Fluorescence Correlation Spectroscopy for Measuring Diffusion and Concentration," ChemPhysChem 6, 2324-2336 (2005).
[CrossRef]

I. Gregor, D. Patra, and J. Enderlein, "Optical Saturation in Fluorescence Correlation Spectroscopy under Continuous-Wave and Pulsed Excitation," ChemPhysChem 6, 164-70 (2005).
[CrossRef]

I. Gregor, D. Patra, and J. Enderlein, "Optical Saturation in Fluorescence Correlation Spectroscopy under Continuous-Wave and Pulsed Excitation," ChemPhysChem 6, 164-170 (2005).
[CrossRef]

B. K. Müller, E. Zaychikov, C. Bräuchle, and D. C. Lamb, "Pulsed interleaved excitation," Biophys. J. 89, 3508-3522 (2005).
[CrossRef]

2004 (2)

O. Haeberlé, "Focusing of light through a stratified medium: a practical approach for computing microscope point spread functions. Part II: confocal and multiphoton microscopy," Opt. Commun. 235, 1 (2004).
[CrossRef]

G. Nishimura and M. Kinjo "Systematic error in fluorescence correlation measurements identified by a simple saturation model of fluorescence," Anal. Chem. 76, 1963-1970 (2004).
[CrossRef]

2003 (4)

2001 (1)

M. Böhmer, F. Pampaloni, M. Wahl, H. J. Rahn, R. Erdmann, and J. Enderlein, "Time-resolved confocal scanning device for ultrasensitive fluorescence detection," Rev. Sci. Instrum. 72, 4145-4152 (2001).
[CrossRef]

2000 (1)

1999 (1)

P. D. Higdon, P. Török, and T. Wilson, "Imaging properties of high aperture multiphoton fluorescence scanning optical microscopes," J. Microsc. 193, 127-141 (1999).
[CrossRef]

1998 (2)

P. Török, P. D. Higdon, and T. Wilson, "Theory for confocal and conventional microscopes imaging small dielectric scatterers," J. Mod. Opt. 45, 1681-1698 (1998).

A. Egner, M. Schrader, and S. W. Hell, "Refractive index mismatch induced intensity and phase variations in fluorescence confocal, multiphoton and 4Pi-microscopy," Opt. Commun. 153, 211 (1998).
[CrossRef]

1997 (1)

1995 (1)

1974 (2)

E. L. Elson and D. Magde "Fluorescence Corelation Spectroscopy I. Conceptual Basis and Theory," Bioploymers 13, 1-27 (1974).
[CrossRef]

D. Magde, E. Elson, and W. W. Webb "Fluorescence Corelation Spectroscopy II. An Experimental Realization," Biopolymers 13, 29-61 (1974).
[CrossRef]

1972 (1)

D. Magde, E. Elson, and W. W. Webb "Thermodynamic fluctuations in a reacting system - measurement by fluorescence correlation spectroscopy," Phys. Rev. Lett. 29, 705-708, (1972).
[CrossRef]

1970 (1)

G. Nomarski, "Interference Microscopy - State of Art and Its Future," J. Opt. Soc. Am. 60, 1575-1575 (1970).

1959 (2)

E. Wolf, "Electromagnetic diffraction in optical systems I. An integral representation of the image field," Proc. Roy. Soc. London A 253, 349-357 (1959).

B. Richards and E. Wolf, "Electromagnetic diffraction in optical systems II. Structure of the image field in an aplanatic system," Proc. Roy. Soc. London A 253, 358-379 (1959).

Ameloot, M.

J. Humpolicková, E. Gielen, A. Benda, V. Fagulova, J. Vercammen, M. VandeVen, M. Hof, M. Ameloot, and Y. Engelborghs, "Probing Diffusion Laws within Cellular Membranes by Z-Scan Fluorescence Correlation Spectroscopy," Biophys. J. Biophys. Lett. 91, L23-L25 (2006).

Ammar, M.

Benda, A.

J. Humpolicková, E. Gielen, A. Benda, V. Fagulova, J. Vercammen, M. VandeVen, M. Hof, M. Ameloot, and Y. Engelborghs, "Probing Diffusion Laws within Cellular Membranes by Z-Scan Fluorescence Correlation Spectroscopy," Biophys. J. Biophys. Lett. 91, L23-L25 (2006).

A. Benda, M. Benes, V. Marecek, A. Lhotsky, W.T. Hermens, and M. Hof, "How To Determine Diffusion Coefficients in Planar Phospholipid Systems by Confocal Fluorescence Correlation Spectroscopy," Langmuir 19, 4120-4126 (2003).
[CrossRef]

Benes, M.

A. Benda, M. Benes, V. Marecek, A. Lhotsky, W.T. Hermens, and M. Hof, "How To Determine Diffusion Coefficients in Planar Phospholipid Systems by Confocal Fluorescence Correlation Spectroscopy," Langmuir 19, 4120-4126 (2003).
[CrossRef]

Berland, K.

Böhmer, M.

M. Böhmer, F. Pampaloni, M. Wahl, H. J. Rahn, R. Erdmann, and J. Enderlein, "Time-resolved confocal scanning device for ultrasensitive fluorescence detection," Rev. Sci. Instrum. 72, 4145-4152 (2001).
[CrossRef]

Booker, J.

Bräuchle, C.

B. K. Müller, E. Zaychikov, C. Bräuchle, and D. C. Lamb, "Pulsed interleaved excitation," Biophys. J. 89, 3508-3522 (2005).
[CrossRef]

Dertinger, T.

A. Loman, T. Dertinger, F. Koberling, and J. Enderlein, "Comparison of optical saturation effects in conventional and dual-focus fluorescence correlation spectroscopy," Chem. Phys. Lett. 459, 18-21 (2008).
[CrossRef]

T. Dertinger, V. Pacheco, I. von der Hocht, R. Hartmann, I. Gregor, and J. Enderlein, "Two-focus fluorescence correlation spectroscopy: A new tool for accurate and absolute diffusion measurements," ChemPhysChem 8, 433-443 (2007).
[CrossRef]

J. Enderlein, I. Gregor, D. Patra, T. Dertinger, and U. B. Kaupp, "Performance of Fluorescence Correlation Spectroscopy for Measuring Diffusion and Concentration," ChemPhysChem 6, 2324-2336 (2005).
[CrossRef]

Donnert, G.

G. Donnert, C. Eggeling, and S. W Hell, "Major signal increase in fluorescence microscopy through dark-state relaxation," Nature Meth. 4, 81-86 (2007).
[CrossRef]

Eggeling, C.

G. Donnert, C. Eggeling, and S. W Hell, "Major signal increase in fluorescence microscopy through dark-state relaxation," Nature Meth. 4, 81-86 (2007).
[CrossRef]

Egner, A.

A. Egner, M. Schrader, and S. W. Hell, "Refractive index mismatch induced intensity and phase variations in fluorescence confocal, multiphoton and 4Pi-microscopy," Opt. Commun. 153, 211 (1998).
[CrossRef]

Elson, E.

D. Magde, E. Elson, and W. W. Webb "Fluorescence Corelation Spectroscopy II. An Experimental Realization," Biopolymers 13, 29-61 (1974).
[CrossRef]

D. Magde, E. Elson, and W. W. Webb "Thermodynamic fluctuations in a reacting system - measurement by fluorescence correlation spectroscopy," Phys. Rev. Lett. 29, 705-708, (1972).
[CrossRef]

Elson, E. L.

E. L. Elson and D. Magde "Fluorescence Corelation Spectroscopy I. Conceptual Basis and Theory," Bioploymers 13, 1-27 (1974).
[CrossRef]

Enderlein, J.

C. B. Müller, K. Wei??, W. Richtering, A. Loman, and J. Enderlein, "Calibrating Differential Interference Contrast Microscopy with dual-focus Fluorescence Correlation Spectroscopy," Opt. Express 16, 4322-9 (2008).
[CrossRef]

C. B. Müller, A. Loman, V. Pacheco, F. Koberling, D. Willbold, W. Richtering, and J. Enderlein, "Precise Measurement of Diffusion by Multi-Color Dual-Focus Fluorescence Correlation Spectroscopy," Eur. Phys. Lett. 83, 46001 (2008).
[CrossRef]

A. Loman, T. Dertinger, F. Koberling, and J. Enderlein, "Comparison of optical saturation effects in conventional and dual-focus fluorescence correlation spectroscopy," Chem. Phys. Lett. 459, 18-21 (2008).
[CrossRef]

T. Dertinger, V. Pacheco, I. von der Hocht, R. Hartmann, I. Gregor, and J. Enderlein, "Two-focus fluorescence correlation spectroscopy: A new tool for accurate and absolute diffusion measurements," ChemPhysChem 8, 433-443 (2007).
[CrossRef]

J. Enderlein, I. Gregor, D. Patra, T. Dertinger, and U. B. Kaupp, "Performance of Fluorescence Correlation Spectroscopy for Measuring Diffusion and Concentration," ChemPhysChem 6, 2324-2336 (2005).
[CrossRef]

I. Gregor and J. Enderlein "Focusing astigmatic Gaussian beams through optical systems with a high numerical aperture," Opt. Lett. 30, 2527-9 (2005).
[CrossRef]

I. Gregor, D. Patra, and J. Enderlein, "Optical Saturation in Fluorescence Correlation Spectroscopy under Continuous-Wave and Pulsed Excitation," ChemPhysChem 6, 164-70 (2005).
[CrossRef]

I. Gregor, D. Patra, and J. Enderlein, "Optical Saturation in Fluorescence Correlation Spectroscopy under Continuous-Wave and Pulsed Excitation," ChemPhysChem 6, 164-170 (2005).
[CrossRef]

M. Wahl, I. Gregor, M. Patting, and J. Enderlein, "Fast calculation of fluorescence correlation data with asynchronous time-correlated single-photon counting," Opt. Express 11, 3583-91 (2003).

M. Böhmer, F. Pampaloni, M. Wahl, H. J. Rahn, R. Erdmann, and J. Enderlein, "Time-resolved confocal scanning device for ultrasensitive fluorescence detection," Rev. Sci. Instrum. 72, 4145-4152 (2001).
[CrossRef]

Engelborghs, Y.

J. Humpolicková, E. Gielen, A. Benda, V. Fagulova, J. Vercammen, M. VandeVen, M. Hof, M. Ameloot, and Y. Engelborghs, "Probing Diffusion Laws within Cellular Membranes by Z-Scan Fluorescence Correlation Spectroscopy," Biophys. J. Biophys. Lett. 91, L23-L25 (2006).

Erdmann, R.

M. Böhmer, F. Pampaloni, M. Wahl, H. J. Rahn, R. Erdmann, and J. Enderlein, "Time-resolved confocal scanning device for ultrasensitive fluorescence detection," Rev. Sci. Instrum. 72, 4145-4152 (2001).
[CrossRef]

Fagulova, V.

J. Humpolicková, E. Gielen, A. Benda, V. Fagulova, J. Vercammen, M. VandeVen, M. Hof, M. Ameloot, and Y. Engelborghs, "Probing Diffusion Laws within Cellular Membranes by Z-Scan Fluorescence Correlation Spectroscopy," Biophys. J. Biophys. Lett. 91, L23-L25 (2006).

Furukawa, H.

Gielen, E.

J. Humpolicková, E. Gielen, A. Benda, V. Fagulova, J. Vercammen, M. VandeVen, M. Hof, M. Ameloot, and Y. Engelborghs, "Probing Diffusion Laws within Cellular Membranes by Z-Scan Fluorescence Correlation Spectroscopy," Biophys. J. Biophys. Lett. 91, L23-L25 (2006).

Gregor, I.

T. Dertinger, V. Pacheco, I. von der Hocht, R. Hartmann, I. Gregor, and J. Enderlein, "Two-focus fluorescence correlation spectroscopy: A new tool for accurate and absolute diffusion measurements," ChemPhysChem 8, 433-443 (2007).
[CrossRef]

J. Enderlein, I. Gregor, D. Patra, T. Dertinger, and U. B. Kaupp, "Performance of Fluorescence Correlation Spectroscopy for Measuring Diffusion and Concentration," ChemPhysChem 6, 2324-2336 (2005).
[CrossRef]

I. Gregor and J. Enderlein "Focusing astigmatic Gaussian beams through optical systems with a high numerical aperture," Opt. Lett. 30, 2527-9 (2005).
[CrossRef]

I. Gregor, D. Patra, and J. Enderlein, "Optical Saturation in Fluorescence Correlation Spectroscopy under Continuous-Wave and Pulsed Excitation," ChemPhysChem 6, 164-170 (2005).
[CrossRef]

I. Gregor, D. Patra, and J. Enderlein, "Optical Saturation in Fluorescence Correlation Spectroscopy under Continuous-Wave and Pulsed Excitation," ChemPhysChem 6, 164-70 (2005).
[CrossRef]

M. Wahl, I. Gregor, M. Patting, and J. Enderlein, "Fast calculation of fluorescence correlation data with asynchronous time-correlated single-photon counting," Opt. Express 11, 3583-91 (2003).

Haeberlé, O.

O. Haeberlé, "Focusing of light through a stratified medium: a practical approach for computing microscope point spread functions. Part II: confocal and multiphoton microscopy," Opt. Commun. 235, 1 (2004).
[CrossRef]

O. Haeberlé, M. Ammar, H. Furukawa, K. Tenjimbayashi, and P. Török, "The point spread function of optical microscopes imaging through stratified media," Opt. Express 11, 2964 (2003).

Hartmann, R.

T. Dertinger, V. Pacheco, I. von der Hocht, R. Hartmann, I. Gregor, and J. Enderlein, "Two-focus fluorescence correlation spectroscopy: A new tool for accurate and absolute diffusion measurements," ChemPhysChem 8, 433-443 (2007).
[CrossRef]

Hell, S. W

G. Donnert, C. Eggeling, and S. W Hell, "Major signal increase in fluorescence microscopy through dark-state relaxation," Nature Meth. 4, 81-86 (2007).
[CrossRef]

Hell, S. W.

A. Egner, M. Schrader, and S. W. Hell, "Refractive index mismatch induced intensity and phase variations in fluorescence confocal, multiphoton and 4Pi-microscopy," Opt. Commun. 153, 211 (1998).
[CrossRef]

Hermens, W.T.

A. Benda, M. Benes, V. Marecek, A. Lhotsky, W.T. Hermens, and M. Hof, "How To Determine Diffusion Coefficients in Planar Phospholipid Systems by Confocal Fluorescence Correlation Spectroscopy," Langmuir 19, 4120-4126 (2003).
[CrossRef]

Higdon, P. D.

P. D. Higdon, P. Török, and T. Wilson, "Imaging properties of high aperture multiphoton fluorescence scanning optical microscopes," J. Microsc. 193, 127-141 (1999).
[CrossRef]

P. Török, P. D. Higdon, and T. Wilson, "Theory for confocal and conventional microscopes imaging small dielectric scatterers," J. Mod. Opt. 45, 1681-1698 (1998).

Hof, M.

J. Humpolicková, E. Gielen, A. Benda, V. Fagulova, J. Vercammen, M. VandeVen, M. Hof, M. Ameloot, and Y. Engelborghs, "Probing Diffusion Laws within Cellular Membranes by Z-Scan Fluorescence Correlation Spectroscopy," Biophys. J. Biophys. Lett. 91, L23-L25 (2006).

A. Benda, M. Benes, V. Marecek, A. Lhotsky, W.T. Hermens, and M. Hof, "How To Determine Diffusion Coefficients in Planar Phospholipid Systems by Confocal Fluorescence Correlation Spectroscopy," Langmuir 19, 4120-4126 (2003).
[CrossRef]

Humpolicková, J.

J. Humpolicková, E. Gielen, A. Benda, V. Fagulova, J. Vercammen, M. VandeVen, M. Hof, M. Ameloot, and Y. Engelborghs, "Probing Diffusion Laws within Cellular Membranes by Z-Scan Fluorescence Correlation Spectroscopy," Biophys. J. Biophys. Lett. 91, L23-L25 (2006).

Kaupp, U. B.

J. Enderlein, I. Gregor, D. Patra, T. Dertinger, and U. B. Kaupp, "Performance of Fluorescence Correlation Spectroscopy for Measuring Diffusion and Concentration," ChemPhysChem 6, 2324-2336 (2005).
[CrossRef]

Kinjo, M.

G. Nishimura and M. Kinjo "Systematic error in fluorescence correlation measurements identified by a simple saturation model of fluorescence," Anal. Chem. 76, 1963-1970 (2004).
[CrossRef]

Koberling, F.

C. B. Müller, A. Loman, V. Pacheco, F. Koberling, D. Willbold, W. Richtering, and J. Enderlein, "Precise Measurement of Diffusion by Multi-Color Dual-Focus Fluorescence Correlation Spectroscopy," Eur. Phys. Lett. 83, 46001 (2008).
[CrossRef]

A. Loman, T. Dertinger, F. Koberling, and J. Enderlein, "Comparison of optical saturation effects in conventional and dual-focus fluorescence correlation spectroscopy," Chem. Phys. Lett. 459, 18-21 (2008).
[CrossRef]

Laczik, G. R.

Lamb, D. C.

B. K. Müller, E. Zaychikov, C. Bräuchle, and D. C. Lamb, "Pulsed interleaved excitation," Biophys. J. 89, 3508-3522 (2005).
[CrossRef]

Lasser, T.

Leitgeb, R. A.

Leutenegger, M.

Lhotsky, A.

A. Benda, M. Benes, V. Marecek, A. Lhotsky, W.T. Hermens, and M. Hof, "How To Determine Diffusion Coefficients in Planar Phospholipid Systems by Confocal Fluorescence Correlation Spectroscopy," Langmuir 19, 4120-4126 (2003).
[CrossRef]

Loman, A.

C. B. Müller, K. Wei??, W. Richtering, A. Loman, and J. Enderlein, "Calibrating Differential Interference Contrast Microscopy with dual-focus Fluorescence Correlation Spectroscopy," Opt. Express 16, 4322-9 (2008).
[CrossRef]

A. Loman, T. Dertinger, F. Koberling, and J. Enderlein, "Comparison of optical saturation effects in conventional and dual-focus fluorescence correlation spectroscopy," Chem. Phys. Lett. 459, 18-21 (2008).
[CrossRef]

C. B. Müller, A. Loman, V. Pacheco, F. Koberling, D. Willbold, W. Richtering, and J. Enderlein, "Precise Measurement of Diffusion by Multi-Color Dual-Focus Fluorescence Correlation Spectroscopy," Eur. Phys. Lett. 83, 46001 (2008).
[CrossRef]

Magde, D.

D. Magde, E. Elson, and W. W. Webb "Fluorescence Corelation Spectroscopy II. An Experimental Realization," Biopolymers 13, 29-61 (1974).
[CrossRef]

E. L. Elson and D. Magde "Fluorescence Corelation Spectroscopy I. Conceptual Basis and Theory," Bioploymers 13, 1-27 (1974).
[CrossRef]

D. Magde, E. Elson, and W. W. Webb "Thermodynamic fluctuations in a reacting system - measurement by fluorescence correlation spectroscopy," Phys. Rev. Lett. 29, 705-708, (1972).
[CrossRef]

Marecek, V.

A. Benda, M. Benes, V. Marecek, A. Lhotsky, W.T. Hermens, and M. Hof, "How To Determine Diffusion Coefficients in Planar Phospholipid Systems by Confocal Fluorescence Correlation Spectroscopy," Langmuir 19, 4120-4126 (2003).
[CrossRef]

Müller, B. K.

B. K. Müller, E. Zaychikov, C. Bräuchle, and D. C. Lamb, "Pulsed interleaved excitation," Biophys. J. 89, 3508-3522 (2005).
[CrossRef]

Müller, C. B.

C. B. Müller, K. Wei??, W. Richtering, A. Loman, and J. Enderlein, "Calibrating Differential Interference Contrast Microscopy with dual-focus Fluorescence Correlation Spectroscopy," Opt. Express 16, 4322-9 (2008).
[CrossRef]

C. B. Müller, A. Loman, V. Pacheco, F. Koberling, D. Willbold, W. Richtering, and J. Enderlein, "Precise Measurement of Diffusion by Multi-Color Dual-Focus Fluorescence Correlation Spectroscopy," Eur. Phys. Lett. 83, 46001 (2008).
[CrossRef]

Munro, P. R. T.

Nishimura, G.

G. Nishimura and M. Kinjo "Systematic error in fluorescence correlation measurements identified by a simple saturation model of fluorescence," Anal. Chem. 76, 1963-1970 (2004).
[CrossRef]

Nomarski, G.

G. Nomarski, "Interference Microscopy - State of Art and Its Future," J. Opt. Soc. Am. 60, 1575-1575 (1970).

Pacheco, V.

C. B. Müller, A. Loman, V. Pacheco, F. Koberling, D. Willbold, W. Richtering, and J. Enderlein, "Precise Measurement of Diffusion by Multi-Color Dual-Focus Fluorescence Correlation Spectroscopy," Eur. Phys. Lett. 83, 46001 (2008).
[CrossRef]

T. Dertinger, V. Pacheco, I. von der Hocht, R. Hartmann, I. Gregor, and J. Enderlein, "Two-focus fluorescence correlation spectroscopy: A new tool for accurate and absolute diffusion measurements," ChemPhysChem 8, 433-443 (2007).
[CrossRef]

Pampaloni, F.

M. Böhmer, F. Pampaloni, M. Wahl, H. J. Rahn, R. Erdmann, and J. Enderlein, "Time-resolved confocal scanning device for ultrasensitive fluorescence detection," Rev. Sci. Instrum. 72, 4145-4152 (2001).
[CrossRef]

Patra, D.

I. Gregor, D. Patra, and J. Enderlein, "Optical Saturation in Fluorescence Correlation Spectroscopy under Continuous-Wave and Pulsed Excitation," ChemPhysChem 6, 164-70 (2005).
[CrossRef]

I. Gregor, D. Patra, and J. Enderlein, "Optical Saturation in Fluorescence Correlation Spectroscopy under Continuous-Wave and Pulsed Excitation," ChemPhysChem 6, 164-170 (2005).
[CrossRef]

J. Enderlein, I. Gregor, D. Patra, T. Dertinger, and U. B. Kaupp, "Performance of Fluorescence Correlation Spectroscopy for Measuring Diffusion and Concentration," ChemPhysChem 6, 2324-2336 (2005).
[CrossRef]

Patting, M.

Petrasek, Z.

Z. Petrasek and P. Schwille, "Precise measurement of diffusion coefficients using scanning fluorescence correlation spectroscopy," Biophys. J. 94, 1437-1448 (2008).
[CrossRef]

Rahn, H. J.

M. Böhmer, F. Pampaloni, M. Wahl, H. J. Rahn, R. Erdmann, and J. Enderlein, "Time-resolved confocal scanning device for ultrasensitive fluorescence detection," Rev. Sci. Instrum. 72, 4145-4152 (2001).
[CrossRef]

Rao, R.

Richards, B.

B. Richards and E. Wolf, "Electromagnetic diffraction in optical systems II. Structure of the image field in an aplanatic system," Proc. Roy. Soc. London A 253, 358-379 (1959).

Richtering, W.

C. B. Müller, A. Loman, V. Pacheco, F. Koberling, D. Willbold, W. Richtering, and J. Enderlein, "Precise Measurement of Diffusion by Multi-Color Dual-Focus Fluorescence Correlation Spectroscopy," Eur. Phys. Lett. 83, 46001 (2008).
[CrossRef]

C. B. Müller, K. Wei??, W. Richtering, A. Loman, and J. Enderlein, "Calibrating Differential Interference Contrast Microscopy with dual-focus Fluorescence Correlation Spectroscopy," Opt. Express 16, 4322-9 (2008).
[CrossRef]

Ries, J.

J. Ries and P. Schwille, "Studying Slow Membrane Dynamics with Continuous Wave Scanning Fluorescence Correlation Spectroscopy," Biophys. J. 91, 1915-1924 (2006).
[CrossRef]

Schrader, M.

A. Egner, M. Schrader, and S. W. Hell, "Refractive index mismatch induced intensity and phase variations in fluorescence confocal, multiphoton and 4Pi-microscopy," Opt. Commun. 153, 211 (1998).
[CrossRef]

Schwille, P.

Z. Petrasek and P. Schwille, "Precise measurement of diffusion coefficients using scanning fluorescence correlation spectroscopy," Biophys. J. 94, 1437-1448 (2008).
[CrossRef]

J. Ries and P. Schwille, "Studying Slow Membrane Dynamics with Continuous Wave Scanning Fluorescence Correlation Spectroscopy," Biophys. J. 91, 1915-1924 (2006).
[CrossRef]

Shen, G.

Tenjimbayashi, K.

Török, P.

VandeVen, M.

J. Humpolicková, E. Gielen, A. Benda, V. Fagulova, J. Vercammen, M. VandeVen, M. Hof, M. Ameloot, and Y. Engelborghs, "Probing Diffusion Laws within Cellular Membranes by Z-Scan Fluorescence Correlation Spectroscopy," Biophys. J. Biophys. Lett. 91, L23-L25 (2006).

Varga, P.

Varga, Z.

Vercammen, J.

J. Humpolicková, E. Gielen, A. Benda, V. Fagulova, J. Vercammen, M. VandeVen, M. Hof, M. Ameloot, and Y. Engelborghs, "Probing Diffusion Laws within Cellular Membranes by Z-Scan Fluorescence Correlation Spectroscopy," Biophys. J. Biophys. Lett. 91, L23-L25 (2006).

von der Hocht, I.

T. Dertinger, V. Pacheco, I. von der Hocht, R. Hartmann, I. Gregor, and J. Enderlein, "Two-focus fluorescence correlation spectroscopy: A new tool for accurate and absolute diffusion measurements," ChemPhysChem 8, 433-443 (2007).
[CrossRef]

Wahl, M.

M. Wahl, I. Gregor, M. Patting, and J. Enderlein, "Fast calculation of fluorescence correlation data with asynchronous time-correlated single-photon counting," Opt. Express 11, 3583-91 (2003).

M. Böhmer, F. Pampaloni, M. Wahl, H. J. Rahn, R. Erdmann, and J. Enderlein, "Time-resolved confocal scanning device for ultrasensitive fluorescence detection," Rev. Sci. Instrum. 72, 4145-4152 (2001).
[CrossRef]

Webb, W. W.

D. Magde, E. Elson, and W. W. Webb "Fluorescence Corelation Spectroscopy II. An Experimental Realization," Biopolymers 13, 29-61 (1974).
[CrossRef]

D. Magde, E. Elson, and W. W. Webb "Thermodynamic fluctuations in a reacting system - measurement by fluorescence correlation spectroscopy," Phys. Rev. Lett. 29, 705-708, (1972).
[CrossRef]

Wei??, K.

Willbold, D.

C. B. Müller, A. Loman, V. Pacheco, F. Koberling, D. Willbold, W. Richtering, and J. Enderlein, "Precise Measurement of Diffusion by Multi-Color Dual-Focus Fluorescence Correlation Spectroscopy," Eur. Phys. Lett. 83, 46001 (2008).
[CrossRef]

Wilson, T.

P. D. Higdon, P. Török, and T. Wilson, "Imaging properties of high aperture multiphoton fluorescence scanning optical microscopes," J. Microsc. 193, 127-141 (1999).
[CrossRef]

P. Török, P. D. Higdon, and T. Wilson, "Theory for confocal and conventional microscopes imaging small dielectric scatterers," J. Mod. Opt. 45, 1681-1698 (1998).

Wolf, E.

E. Wolf, "Electromagnetic diffraction in optical systems I. An integral representation of the image field," Proc. Roy. Soc. London A 253, 349-357 (1959).

B. Richards and E. Wolf, "Electromagnetic diffraction in optical systems II. Structure of the image field in an aplanatic system," Proc. Roy. Soc. London A 253, 358-379 (1959).

Zaychikov, E.

B. K. Müller, E. Zaychikov, C. Bräuchle, and D. C. Lamb, "Pulsed interleaved excitation," Biophys. J. 89, 3508-3522 (2005).
[CrossRef]

Anal. Chem. (1)

G. Nishimura and M. Kinjo "Systematic error in fluorescence correlation measurements identified by a simple saturation model of fluorescence," Anal. Chem. 76, 1963-1970 (2004).
[CrossRef]

Appl. Opt. (2)

Biophys. J. (3)

J. Ries and P. Schwille, "Studying Slow Membrane Dynamics with Continuous Wave Scanning Fluorescence Correlation Spectroscopy," Biophys. J. 91, 1915-1924 (2006).
[CrossRef]

Z. Petrasek and P. Schwille, "Precise measurement of diffusion coefficients using scanning fluorescence correlation spectroscopy," Biophys. J. 94, 1437-1448 (2008).
[CrossRef]

B. K. Müller, E. Zaychikov, C. Bräuchle, and D. C. Lamb, "Pulsed interleaved excitation," Biophys. J. 89, 3508-3522 (2005).
[CrossRef]

Biophys. J. Biophys. Lett. (1)

J. Humpolicková, E. Gielen, A. Benda, V. Fagulova, J. Vercammen, M. VandeVen, M. Hof, M. Ameloot, and Y. Engelborghs, "Probing Diffusion Laws within Cellular Membranes by Z-Scan Fluorescence Correlation Spectroscopy," Biophys. J. Biophys. Lett. 91, L23-L25 (2006).

Bioploymers (1)

E. L. Elson and D. Magde "Fluorescence Corelation Spectroscopy I. Conceptual Basis and Theory," Bioploymers 13, 1-27 (1974).
[CrossRef]

Biopolymers (1)

D. Magde, E. Elson, and W. W. Webb "Fluorescence Corelation Spectroscopy II. An Experimental Realization," Biopolymers 13, 29-61 (1974).
[CrossRef]

Chem. Phys. Lett. (1)

A. Loman, T. Dertinger, F. Koberling, and J. Enderlein, "Comparison of optical saturation effects in conventional and dual-focus fluorescence correlation spectroscopy," Chem. Phys. Lett. 459, 18-21 (2008).
[CrossRef]

ChemPhysChem (4)

T. Dertinger, V. Pacheco, I. von der Hocht, R. Hartmann, I. Gregor, and J. Enderlein, "Two-focus fluorescence correlation spectroscopy: A new tool for accurate and absolute diffusion measurements," ChemPhysChem 8, 433-443 (2007).
[CrossRef]

J. Enderlein, I. Gregor, D. Patra, T. Dertinger, and U. B. Kaupp, "Performance of Fluorescence Correlation Spectroscopy for Measuring Diffusion and Concentration," ChemPhysChem 6, 2324-2336 (2005).
[CrossRef]

I. Gregor, D. Patra, and J. Enderlein, "Optical Saturation in Fluorescence Correlation Spectroscopy under Continuous-Wave and Pulsed Excitation," ChemPhysChem 6, 164-70 (2005).
[CrossRef]

I. Gregor, D. Patra, and J. Enderlein, "Optical Saturation in Fluorescence Correlation Spectroscopy under Continuous-Wave and Pulsed Excitation," ChemPhysChem 6, 164-170 (2005).
[CrossRef]

Eur. Phys. Lett. (1)

C. B. Müller, A. Loman, V. Pacheco, F. Koberling, D. Willbold, W. Richtering, and J. Enderlein, "Precise Measurement of Diffusion by Multi-Color Dual-Focus Fluorescence Correlation Spectroscopy," Eur. Phys. Lett. 83, 46001 (2008).
[CrossRef]

J. Microsc. (1)

P. D. Higdon, P. Török, and T. Wilson, "Imaging properties of high aperture multiphoton fluorescence scanning optical microscopes," J. Microsc. 193, 127-141 (1999).
[CrossRef]

J. Mod. Opt. (1)

P. Török, P. D. Higdon, and T. Wilson, "Theory for confocal and conventional microscopes imaging small dielectric scatterers," J. Mod. Opt. 45, 1681-1698 (1998).

J. Opt. Soc. Am. (1)

G. Nomarski, "Interference Microscopy - State of Art and Its Future," J. Opt. Soc. Am. 60, 1575-1575 (1970).

J. Opt. Soc. Am. A (1)

Langmuir (1)

A. Benda, M. Benes, V. Marecek, A. Lhotsky, W.T. Hermens, and M. Hof, "How To Determine Diffusion Coefficients in Planar Phospholipid Systems by Confocal Fluorescence Correlation Spectroscopy," Langmuir 19, 4120-4126 (2003).
[CrossRef]

Nature Meth. (1)

G. Donnert, C. Eggeling, and S. W Hell, "Major signal increase in fluorescence microscopy through dark-state relaxation," Nature Meth. 4, 81-86 (2007).
[CrossRef]

Opt. Commun. (2)

A. Egner, M. Schrader, and S. W. Hell, "Refractive index mismatch induced intensity and phase variations in fluorescence confocal, multiphoton and 4Pi-microscopy," Opt. Commun. 153, 211 (1998).
[CrossRef]

O. Haeberlé, "Focusing of light through a stratified medium: a practical approach for computing microscope point spread functions. Part II: confocal and multiphoton microscopy," Opt. Commun. 235, 1 (2004).
[CrossRef]

Opt. Express (6)

Opt. Lett. (2)

Phys. Rev. Lett. (1)

D. Magde, E. Elson, and W. W. Webb "Thermodynamic fluctuations in a reacting system - measurement by fluorescence correlation spectroscopy," Phys. Rev. Lett. 29, 705-708, (1972).
[CrossRef]

Proc. Roy. Soc. London A (2)

E. Wolf, "Electromagnetic diffraction in optical systems I. An integral representation of the image field," Proc. Roy. Soc. London A 253, 349-357 (1959).

B. Richards and E. Wolf, "Electromagnetic diffraction in optical systems II. Structure of the image field in an aplanatic system," Proc. Roy. Soc. London A 253, 358-379 (1959).

Rev. Sci. Instrum. (1)

M. Böhmer, F. Pampaloni, M. Wahl, H. J. Rahn, R. Erdmann, and J. Enderlein, "Time-resolved confocal scanning device for ultrasensitive fluorescence detection," Rev. Sci. Instrum. 72, 4145-4152 (2001).
[CrossRef]

Other (2)

J. Widengren and ??. Mets, Single-Molecule Detection in Solution - Methods and Applications, Eds. C. Zander, J. Enderlein, and R. A. Keller (Wiley-VCH, 2002) pp. 69-95.

R. Rigler and E. Elson, Eds. Fluorescence Correlation Spectroscopy (Springer, 2001).

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