Abstract

Three-dimensional (3D) spatial information can be encoded in two-dimensional images of fluorescent nanoparticles by astigmatic imaging. We combined this method with light sheet microscopy for high contrast single particle imaging up to 200 µm deep within living tissue and real-time image analysis to determine 3D particle localizations with nanometer precision and millisecond temporal resolution. Axial information was instantly directed to the sample stage to keep a moving particle within the focal plane in an active feedback loop. We demonstrated 3D tracking of nanoparticles at an unprecedented depth throughout large cell nuclei over several thousand frames and a range of more than 10 µm in each spatial dimension, while simultaneously acquiring optically sectioned wide field images. We conclude that this 3D particle tracking technique employing light sheet microscopy presents a valuable extension to the nanoscopy toolbox.

© 2012 OSA

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  1. M. A. Thompson, J. M. Casolari, M. Badieirostami, P. O. Brown, and W. E. Moerner, “Three-dimensional tracking of single mRNA particles in Saccharomyces cerevisiae using a double-helix point spread function,” Proc. Natl. Acad. Sci. U.S.A.107(42), 17864–17871 (2010).
    [CrossRef] [PubMed]
  2. J. P. Siebrasse, R. Veith, A. Dobay, H. Leonhardt, B. Daneholt, and U. Kubitscheck, “Discontinuous movement of mRNP particles in nucleoplasmic regions devoid of chromatin,” Proc. Natl. Acad. Sci. U.S.A.105(51), 20291–20296 (2008).
    [CrossRef] [PubMed]
  3. N. Arhel, A. Genovesio, K.-A. Kim, S. Miko, E. Perret, J.-C. Olivo-Marin, S. Shorte, and P. Charneau, “Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes,” Nat. Methods3(10), 817–824 (2006).
    [CrossRef] [PubMed]
  4. M. J. Saxton and K. Jacobson, “Single-particle tracking: applications to membrane dynamics,” Annu. Rev. Biophys. Biomol. Struct.26(1), 373–399 (1997).
    [CrossRef] [PubMed]
  5. M. A. Thompson, M. D. Lew, M. Badieirostami, and W. E. Moerner, “Localizing and tracking single nanoscale emitters in three dimensions with high spatiotemporal resolution using a double-helix point spread function,” Nano Lett.10(1), 211–218 (2010).
    [CrossRef] [PubMed]
  6. H. P. Kao and A. S. Verkman, “Tracking of single fluorescent particles in three dimensions: use of cylindrical optics to encode particle position,” Biophys. J.67(3), 1291–1300 (1994).
    [CrossRef] [PubMed]
  7. Y. Katayama, O. Burkacky, M. Meyer, C. Bräuchle, E. Gratton, and D. C. Lamb, “Real-time nanomicroscopy via three-dimensional single-particle tracking,” ChemPhysChem10(14), 2458–2464 (2009).
    [CrossRef] [PubMed]
  8. G. A. Lessard, P. M. Goodwin, and J. H. Werner, “Three-dimensional tracking of individual quantum dots,ˮ,” Appl. Phys. Lett.91(22), 224106 (2007).
    [CrossRef]
  9. K. McHale, A. J. Berglund, and H. Mabuchi, “Quantum dot photon statistics measured by three-dimensional particle tracking,” Nano Lett.7(11), 3535–3539 (2007).
    [CrossRef] [PubMed]
  10. N. P. Wells, G. A. Lessard, P. M. Goodwin, M. E. Phipps, P. J. Cutler, D. S. Lidke, B. S. Wilson, and J. H. Werner, “Time-resolved three-dimensional molecular tracking in live cells,” Nano Lett.10(11), 4732–4737 (2010).
    [CrossRef] [PubMed]
  11. M. D. McMahon, A. J. Berglund, P. Carmichael, J. J. McClelland, and J. A. Liddle, “3D particle trajectories observed by orthogonal tracking microscopy,” ACS Nano3(3), 609–614 (2009).
    [CrossRef] [PubMed]
  12. E. Toprak, H. Balci, B. H. Blehm, and P. R. Selvin, “Three-dimensional particle tracking via bifocal imaging,” Nano Lett.7(7), 2043–2045 (2007).
    [CrossRef] [PubMed]
  13. S. Ram, P. Prabhat, J. Chao, E. S. Ward, and R. J. Ober, “High accuracy 3D quantum dot tracking with multifocal plane microscopy for the study of fast intracellular dynamics in live cells,” Biophys. J.95(12), 6025–6043 (2008).
    [CrossRef] [PubMed]
  14. Y. Sun, J. D. McKenna, J. M. Murray, E. M. Ostap, and Y. E. Goldman, “Parallax: high accuracy three-dimensional single molecule tracking using split images,” Nano Lett.9(7), 2676–2682 (2009).
    [CrossRef] [PubMed]
  15. G. Bolognesi, S. Bianchi, and R. Di Leonardo, “Digital holographic tracking of microprobes for multipoint viscosity measurements,” Opt. Express19(20), 19245–19254 (2011).
    [CrossRef] [PubMed]
  16. L. Holtzer, T. Meckel, and T. Schmidt, “Nanometric three-dimensional tracking of individual quantum dots in cells,ˮ,” Appl. Phys. Lett.90(5), 053902 (2007).
    [CrossRef]
  17. D. Ernst, S. Hain, and J. Köhler, “Setup for single-particle orbit tracking: artifacts and corrections,” J. Opt. Soc. Am. A29(7), 1277–1287 (2012).
    [CrossRef] [PubMed]
  18. M. F. Juette and J. Bewersdorf, “Three-dimensional tracking of single fluorescent particles with submillisecond temporal resolution,” Nano Lett.10(11), 4657–4663 (2010).
    [CrossRef] [PubMed]
  19. J. G. Ritter, R. Veith, A. Veenendaal, J. P. Siebrasse, and U. Kubitscheck, “Light sheet microscopy for single molecule tracking in living tissue,” PLoS ONE5(7), e11639 (2010).
    [CrossRef] [PubMed]
  20. J. P. Siebrasse, T. Kaminski, and U. Kubitscheck, “Nuclear export of single native mRNA molecules observed by light sheet fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.109(24), 9426–9431 (2012).
    [CrossRef] [PubMed]
  21. J. G. Ritter, R. Veith, J.-P. Siebrasse, and U. Kubitscheck, “High-contrast single-particle tracking by selective focal plane illumination microscopy,” Opt. Express16(10), 7142–7152 (2008).
    [CrossRef] [PubMed]
  22. Y. Deng and J. W. Shaevitz, “Effect of aberration on height calibration in three-dimensional localization-based microscopy and particle tracking,” Appl. Opt.48(10), 1886–1890 (2009).
    [CrossRef] [PubMed]
  23. J. G. Ritter, J.-H. Spille, T. Kaminski, and U. Kubitscheck, “A cylindrical zoom lens unit for adjustable optical sectioning in light sheet microscopy,” Biomed. Opt. Express2(1), 185–193 (2011).
    [CrossRef] [PubMed]
  24. J. Huisken and D. Y. Stainier, “Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM),” Opt. Lett.32(17), 2608–2610 (2007).
    [CrossRef] [PubMed]
  25. M. K. Cheezum, W. F. Walker, and W. H. Guilford, “Quantitative comparison of algorithms for tracking single fluorescent particles,” Biophys. J.81(4), 2378–2388 (2001).
    [CrossRef] [PubMed]
  26. I. Izeddin, M. El Beheiry, J. Andilla, D. Ciepielewski, X. Darzacq, and M. Dahan, “PSF shaping using adaptive optics for three-dimensional single-molecule super-resolution imaging and tracking,” Opt. Express20(5), 4957–4967 (2012).
    [CrossRef] [PubMed]
  27. H. Deschout, K. Neyts, and K. Braeckmans, “The influence of movement on the localization precision of sub-resolution particles in fluorescence microscopy,” J Biophotonics5(1), 97–109 (2012).
    [CrossRef] [PubMed]
  28. M. J. Mlodzianoski, M. F. Juette, G. L. Beane, and J. Bewersdorf, “Experimental characterization of 3D localization techniques for particle-tracking and super-resolution microscopy,” Opt. Express17(10), 8264–8277 (2009).
    [CrossRef] [PubMed]
  29. C. von Middendorff, A. Egner, C. Geisler, S. W. Hell, and A. Schönle, “Isotropic 3D nanoscopy based on single emitter switching,” Opt. Express16(25), 20774–20788 (2008).
    [CrossRef] [PubMed]
  30. R. Veith, T. Sorkalla, E. Baumgart, J. Anzt, H. Häberlein, S. Tyagi, J. P. Siebrasse, and U. Kubitscheck, “Balbiani ring mRNPs diffuse through and bind to clusters of large intranuclear molecular structures,” Biophys. J.99(8), 2676–2685 (2010).
    [CrossRef] [PubMed]
  31. D. Ernst, M. Hellmann, J. Kohler, and M. Weiss, “Fractional Brownian motion in crowded fluids,ˮ,” Soft Matter8(18), 4886–4889 (2012).
    [CrossRef]
  32. F. Cella Zanacchi, Z. Lavagnino, M. Perrone Donnorso, A. Del Bue, L. Furia, M. Faretta, and A. Diaspro, “Live-cell 3D super-resolution imaging in thick biological samples,” Nat. Methods8(12), 1047–1049 (2011).
    [CrossRef] [PubMed]
  33. A. P. Fields and A. E. Cohen, “Electrokinetic trapping at the one nanometer limit,” Proc. Natl. Acad. Sci. U.S.A.108(22), 8937–8942 (2011).
    [CrossRef] [PubMed]

2012

J. P. Siebrasse, T. Kaminski, and U. Kubitscheck, “Nuclear export of single native mRNA molecules observed by light sheet fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.109(24), 9426–9431 (2012).
[CrossRef] [PubMed]

D. Ernst, M. Hellmann, J. Kohler, and M. Weiss, “Fractional Brownian motion in crowded fluids,ˮ,” Soft Matter8(18), 4886–4889 (2012).
[CrossRef]

H. Deschout, K. Neyts, and K. Braeckmans, “The influence of movement on the localization precision of sub-resolution particles in fluorescence microscopy,” J Biophotonics5(1), 97–109 (2012).
[CrossRef] [PubMed]

I. Izeddin, M. El Beheiry, J. Andilla, D. Ciepielewski, X. Darzacq, and M. Dahan, “PSF shaping using adaptive optics for three-dimensional single-molecule super-resolution imaging and tracking,” Opt. Express20(5), 4957–4967 (2012).
[CrossRef] [PubMed]

D. Ernst, S. Hain, and J. Köhler, “Setup for single-particle orbit tracking: artifacts and corrections,” J. Opt. Soc. Am. A29(7), 1277–1287 (2012).
[CrossRef] [PubMed]

2011

J. G. Ritter, J.-H. Spille, T. Kaminski, and U. Kubitscheck, “A cylindrical zoom lens unit for adjustable optical sectioning in light sheet microscopy,” Biomed. Opt. Express2(1), 185–193 (2011).
[CrossRef] [PubMed]

G. Bolognesi, S. Bianchi, and R. Di Leonardo, “Digital holographic tracking of microprobes for multipoint viscosity measurements,” Opt. Express19(20), 19245–19254 (2011).
[CrossRef] [PubMed]

F. Cella Zanacchi, Z. Lavagnino, M. Perrone Donnorso, A. Del Bue, L. Furia, M. Faretta, and A. Diaspro, “Live-cell 3D super-resolution imaging in thick biological samples,” Nat. Methods8(12), 1047–1049 (2011).
[CrossRef] [PubMed]

A. P. Fields and A. E. Cohen, “Electrokinetic trapping at the one nanometer limit,” Proc. Natl. Acad. Sci. U.S.A.108(22), 8937–8942 (2011).
[CrossRef] [PubMed]

2010

R. Veith, T. Sorkalla, E. Baumgart, J. Anzt, H. Häberlein, S. Tyagi, J. P. Siebrasse, and U. Kubitscheck, “Balbiani ring mRNPs diffuse through and bind to clusters of large intranuclear molecular structures,” Biophys. J.99(8), 2676–2685 (2010).
[CrossRef] [PubMed]

M. F. Juette and J. Bewersdorf, “Three-dimensional tracking of single fluorescent particles with submillisecond temporal resolution,” Nano Lett.10(11), 4657–4663 (2010).
[CrossRef] [PubMed]

J. G. Ritter, R. Veith, A. Veenendaal, J. P. Siebrasse, and U. Kubitscheck, “Light sheet microscopy for single molecule tracking in living tissue,” PLoS ONE5(7), e11639 (2010).
[CrossRef] [PubMed]

N. P. Wells, G. A. Lessard, P. M. Goodwin, M. E. Phipps, P. J. Cutler, D. S. Lidke, B. S. Wilson, and J. H. Werner, “Time-resolved three-dimensional molecular tracking in live cells,” Nano Lett.10(11), 4732–4737 (2010).
[CrossRef] [PubMed]

M. A. Thompson, J. M. Casolari, M. Badieirostami, P. O. Brown, and W. E. Moerner, “Three-dimensional tracking of single mRNA particles in Saccharomyces cerevisiae using a double-helix point spread function,” Proc. Natl. Acad. Sci. U.S.A.107(42), 17864–17871 (2010).
[CrossRef] [PubMed]

M. A. Thompson, M. D. Lew, M. Badieirostami, and W. E. Moerner, “Localizing and tracking single nanoscale emitters in three dimensions with high spatiotemporal resolution using a double-helix point spread function,” Nano Lett.10(1), 211–218 (2010).
[CrossRef] [PubMed]

2009

Y. Katayama, O. Burkacky, M. Meyer, C. Bräuchle, E. Gratton, and D. C. Lamb, “Real-time nanomicroscopy via three-dimensional single-particle tracking,” ChemPhysChem10(14), 2458–2464 (2009).
[CrossRef] [PubMed]

M. D. McMahon, A. J. Berglund, P. Carmichael, J. J. McClelland, and J. A. Liddle, “3D particle trajectories observed by orthogonal tracking microscopy,” ACS Nano3(3), 609–614 (2009).
[CrossRef] [PubMed]

Y. Sun, J. D. McKenna, J. M. Murray, E. M. Ostap, and Y. E. Goldman, “Parallax: high accuracy three-dimensional single molecule tracking using split images,” Nano Lett.9(7), 2676–2682 (2009).
[CrossRef] [PubMed]

Y. Deng and J. W. Shaevitz, “Effect of aberration on height calibration in three-dimensional localization-based microscopy and particle tracking,” Appl. Opt.48(10), 1886–1890 (2009).
[CrossRef] [PubMed]

M. J. Mlodzianoski, M. F. Juette, G. L. Beane, and J. Bewersdorf, “Experimental characterization of 3D localization techniques for particle-tracking and super-resolution microscopy,” Opt. Express17(10), 8264–8277 (2009).
[CrossRef] [PubMed]

2008

J. G. Ritter, R. Veith, J.-P. Siebrasse, and U. Kubitscheck, “High-contrast single-particle tracking by selective focal plane illumination microscopy,” Opt. Express16(10), 7142–7152 (2008).
[CrossRef] [PubMed]

C. von Middendorff, A. Egner, C. Geisler, S. W. Hell, and A. Schönle, “Isotropic 3D nanoscopy based on single emitter switching,” Opt. Express16(25), 20774–20788 (2008).
[CrossRef] [PubMed]

S. Ram, P. Prabhat, J. Chao, E. S. Ward, and R. J. Ober, “High accuracy 3D quantum dot tracking with multifocal plane microscopy for the study of fast intracellular dynamics in live cells,” Biophys. J.95(12), 6025–6043 (2008).
[CrossRef] [PubMed]

J. P. Siebrasse, R. Veith, A. Dobay, H. Leonhardt, B. Daneholt, and U. Kubitscheck, “Discontinuous movement of mRNP particles in nucleoplasmic regions devoid of chromatin,” Proc. Natl. Acad. Sci. U.S.A.105(51), 20291–20296 (2008).
[CrossRef] [PubMed]

2007

G. A. Lessard, P. M. Goodwin, and J. H. Werner, “Three-dimensional tracking of individual quantum dots,ˮ,” Appl. Phys. Lett.91(22), 224106 (2007).
[CrossRef]

K. McHale, A. J. Berglund, and H. Mabuchi, “Quantum dot photon statistics measured by three-dimensional particle tracking,” Nano Lett.7(11), 3535–3539 (2007).
[CrossRef] [PubMed]

E. Toprak, H. Balci, B. H. Blehm, and P. R. Selvin, “Three-dimensional particle tracking via bifocal imaging,” Nano Lett.7(7), 2043–2045 (2007).
[CrossRef] [PubMed]

L. Holtzer, T. Meckel, and T. Schmidt, “Nanometric three-dimensional tracking of individual quantum dots in cells,ˮ,” Appl. Phys. Lett.90(5), 053902 (2007).
[CrossRef]

J. Huisken and D. Y. Stainier, “Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM),” Opt. Lett.32(17), 2608–2610 (2007).
[CrossRef] [PubMed]

2006

N. Arhel, A. Genovesio, K.-A. Kim, S. Miko, E. Perret, J.-C. Olivo-Marin, S. Shorte, and P. Charneau, “Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes,” Nat. Methods3(10), 817–824 (2006).
[CrossRef] [PubMed]

2001

M. K. Cheezum, W. F. Walker, and W. H. Guilford, “Quantitative comparison of algorithms for tracking single fluorescent particles,” Biophys. J.81(4), 2378–2388 (2001).
[CrossRef] [PubMed]

1997

M. J. Saxton and K. Jacobson, “Single-particle tracking: applications to membrane dynamics,” Annu. Rev. Biophys. Biomol. Struct.26(1), 373–399 (1997).
[CrossRef] [PubMed]

1994

H. P. Kao and A. S. Verkman, “Tracking of single fluorescent particles in three dimensions: use of cylindrical optics to encode particle position,” Biophys. J.67(3), 1291–1300 (1994).
[CrossRef] [PubMed]

Andilla, J.

Anzt, J.

R. Veith, T. Sorkalla, E. Baumgart, J. Anzt, H. Häberlein, S. Tyagi, J. P. Siebrasse, and U. Kubitscheck, “Balbiani ring mRNPs diffuse through and bind to clusters of large intranuclear molecular structures,” Biophys. J.99(8), 2676–2685 (2010).
[CrossRef] [PubMed]

Arhel, N.

N. Arhel, A. Genovesio, K.-A. Kim, S. Miko, E. Perret, J.-C. Olivo-Marin, S. Shorte, and P. Charneau, “Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes,” Nat. Methods3(10), 817–824 (2006).
[CrossRef] [PubMed]

Badieirostami, M.

M. A. Thompson, J. M. Casolari, M. Badieirostami, P. O. Brown, and W. E. Moerner, “Three-dimensional tracking of single mRNA particles in Saccharomyces cerevisiae using a double-helix point spread function,” Proc. Natl. Acad. Sci. U.S.A.107(42), 17864–17871 (2010).
[CrossRef] [PubMed]

M. A. Thompson, M. D. Lew, M. Badieirostami, and W. E. Moerner, “Localizing and tracking single nanoscale emitters in three dimensions with high spatiotemporal resolution using a double-helix point spread function,” Nano Lett.10(1), 211–218 (2010).
[CrossRef] [PubMed]

Balci, H.

E. Toprak, H. Balci, B. H. Blehm, and P. R. Selvin, “Three-dimensional particle tracking via bifocal imaging,” Nano Lett.7(7), 2043–2045 (2007).
[CrossRef] [PubMed]

Baumgart, E.

R. Veith, T. Sorkalla, E. Baumgart, J. Anzt, H. Häberlein, S. Tyagi, J. P. Siebrasse, and U. Kubitscheck, “Balbiani ring mRNPs diffuse through and bind to clusters of large intranuclear molecular structures,” Biophys. J.99(8), 2676–2685 (2010).
[CrossRef] [PubMed]

Beane, G. L.

Berglund, A. J.

M. D. McMahon, A. J. Berglund, P. Carmichael, J. J. McClelland, and J. A. Liddle, “3D particle trajectories observed by orthogonal tracking microscopy,” ACS Nano3(3), 609–614 (2009).
[CrossRef] [PubMed]

K. McHale, A. J. Berglund, and H. Mabuchi, “Quantum dot photon statistics measured by three-dimensional particle tracking,” Nano Lett.7(11), 3535–3539 (2007).
[CrossRef] [PubMed]

Bewersdorf, J.

M. F. Juette and J. Bewersdorf, “Three-dimensional tracking of single fluorescent particles with submillisecond temporal resolution,” Nano Lett.10(11), 4657–4663 (2010).
[CrossRef] [PubMed]

M. J. Mlodzianoski, M. F. Juette, G. L. Beane, and J. Bewersdorf, “Experimental characterization of 3D localization techniques for particle-tracking and super-resolution microscopy,” Opt. Express17(10), 8264–8277 (2009).
[CrossRef] [PubMed]

Bianchi, S.

Blehm, B. H.

E. Toprak, H. Balci, B. H. Blehm, and P. R. Selvin, “Three-dimensional particle tracking via bifocal imaging,” Nano Lett.7(7), 2043–2045 (2007).
[CrossRef] [PubMed]

Bolognesi, G.

Braeckmans, K.

H. Deschout, K. Neyts, and K. Braeckmans, “The influence of movement on the localization precision of sub-resolution particles in fluorescence microscopy,” J Biophotonics5(1), 97–109 (2012).
[CrossRef] [PubMed]

Bräuchle, C.

Y. Katayama, O. Burkacky, M. Meyer, C. Bräuchle, E. Gratton, and D. C. Lamb, “Real-time nanomicroscopy via three-dimensional single-particle tracking,” ChemPhysChem10(14), 2458–2464 (2009).
[CrossRef] [PubMed]

Brown, P. O.

M. A. Thompson, J. M. Casolari, M. Badieirostami, P. O. Brown, and W. E. Moerner, “Three-dimensional tracking of single mRNA particles in Saccharomyces cerevisiae using a double-helix point spread function,” Proc. Natl. Acad. Sci. U.S.A.107(42), 17864–17871 (2010).
[CrossRef] [PubMed]

Burkacky, O.

Y. Katayama, O. Burkacky, M. Meyer, C. Bräuchle, E. Gratton, and D. C. Lamb, “Real-time nanomicroscopy via three-dimensional single-particle tracking,” ChemPhysChem10(14), 2458–2464 (2009).
[CrossRef] [PubMed]

Carmichael, P.

M. D. McMahon, A. J. Berglund, P. Carmichael, J. J. McClelland, and J. A. Liddle, “3D particle trajectories observed by orthogonal tracking microscopy,” ACS Nano3(3), 609–614 (2009).
[CrossRef] [PubMed]

Casolari, J. M.

M. A. Thompson, J. M. Casolari, M. Badieirostami, P. O. Brown, and W. E. Moerner, “Three-dimensional tracking of single mRNA particles in Saccharomyces cerevisiae using a double-helix point spread function,” Proc. Natl. Acad. Sci. U.S.A.107(42), 17864–17871 (2010).
[CrossRef] [PubMed]

Cella Zanacchi, F.

F. Cella Zanacchi, Z. Lavagnino, M. Perrone Donnorso, A. Del Bue, L. Furia, M. Faretta, and A. Diaspro, “Live-cell 3D super-resolution imaging in thick biological samples,” Nat. Methods8(12), 1047–1049 (2011).
[CrossRef] [PubMed]

Chao, J.

S. Ram, P. Prabhat, J. Chao, E. S. Ward, and R. J. Ober, “High accuracy 3D quantum dot tracking with multifocal plane microscopy for the study of fast intracellular dynamics in live cells,” Biophys. J.95(12), 6025–6043 (2008).
[CrossRef] [PubMed]

Charneau, P.

N. Arhel, A. Genovesio, K.-A. Kim, S. Miko, E. Perret, J.-C. Olivo-Marin, S. Shorte, and P. Charneau, “Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes,” Nat. Methods3(10), 817–824 (2006).
[CrossRef] [PubMed]

Cheezum, M. K.

M. K. Cheezum, W. F. Walker, and W. H. Guilford, “Quantitative comparison of algorithms for tracking single fluorescent particles,” Biophys. J.81(4), 2378–2388 (2001).
[CrossRef] [PubMed]

Ciepielewski, D.

Cohen, A. E.

A. P. Fields and A. E. Cohen, “Electrokinetic trapping at the one nanometer limit,” Proc. Natl. Acad. Sci. U.S.A.108(22), 8937–8942 (2011).
[CrossRef] [PubMed]

Cutler, P. J.

N. P. Wells, G. A. Lessard, P. M. Goodwin, M. E. Phipps, P. J. Cutler, D. S. Lidke, B. S. Wilson, and J. H. Werner, “Time-resolved three-dimensional molecular tracking in live cells,” Nano Lett.10(11), 4732–4737 (2010).
[CrossRef] [PubMed]

Dahan, M.

Daneholt, B.

J. P. Siebrasse, R. Veith, A. Dobay, H. Leonhardt, B. Daneholt, and U. Kubitscheck, “Discontinuous movement of mRNP particles in nucleoplasmic regions devoid of chromatin,” Proc. Natl. Acad. Sci. U.S.A.105(51), 20291–20296 (2008).
[CrossRef] [PubMed]

Darzacq, X.

Del Bue, A.

F. Cella Zanacchi, Z. Lavagnino, M. Perrone Donnorso, A. Del Bue, L. Furia, M. Faretta, and A. Diaspro, “Live-cell 3D super-resolution imaging in thick biological samples,” Nat. Methods8(12), 1047–1049 (2011).
[CrossRef] [PubMed]

Deng, Y.

Deschout, H.

H. Deschout, K. Neyts, and K. Braeckmans, “The influence of movement on the localization precision of sub-resolution particles in fluorescence microscopy,” J Biophotonics5(1), 97–109 (2012).
[CrossRef] [PubMed]

Di Leonardo, R.

Diaspro, A.

F. Cella Zanacchi, Z. Lavagnino, M. Perrone Donnorso, A. Del Bue, L. Furia, M. Faretta, and A. Diaspro, “Live-cell 3D super-resolution imaging in thick biological samples,” Nat. Methods8(12), 1047–1049 (2011).
[CrossRef] [PubMed]

Dobay, A.

J. P. Siebrasse, R. Veith, A. Dobay, H. Leonhardt, B. Daneholt, and U. Kubitscheck, “Discontinuous movement of mRNP particles in nucleoplasmic regions devoid of chromatin,” Proc. Natl. Acad. Sci. U.S.A.105(51), 20291–20296 (2008).
[CrossRef] [PubMed]

Egner, A.

El Beheiry, M.

Ernst, D.

D. Ernst, S. Hain, and J. Köhler, “Setup for single-particle orbit tracking: artifacts and corrections,” J. Opt. Soc. Am. A29(7), 1277–1287 (2012).
[CrossRef] [PubMed]

D. Ernst, M. Hellmann, J. Kohler, and M. Weiss, “Fractional Brownian motion in crowded fluids,ˮ,” Soft Matter8(18), 4886–4889 (2012).
[CrossRef]

Faretta, M.

F. Cella Zanacchi, Z. Lavagnino, M. Perrone Donnorso, A. Del Bue, L. Furia, M. Faretta, and A. Diaspro, “Live-cell 3D super-resolution imaging in thick biological samples,” Nat. Methods8(12), 1047–1049 (2011).
[CrossRef] [PubMed]

Fields, A. P.

A. P. Fields and A. E. Cohen, “Electrokinetic trapping at the one nanometer limit,” Proc. Natl. Acad. Sci. U.S.A.108(22), 8937–8942 (2011).
[CrossRef] [PubMed]

Furia, L.

F. Cella Zanacchi, Z. Lavagnino, M. Perrone Donnorso, A. Del Bue, L. Furia, M. Faretta, and A. Diaspro, “Live-cell 3D super-resolution imaging in thick biological samples,” Nat. Methods8(12), 1047–1049 (2011).
[CrossRef] [PubMed]

Geisler, C.

Genovesio, A.

N. Arhel, A. Genovesio, K.-A. Kim, S. Miko, E. Perret, J.-C. Olivo-Marin, S. Shorte, and P. Charneau, “Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes,” Nat. Methods3(10), 817–824 (2006).
[CrossRef] [PubMed]

Goldman, Y. E.

Y. Sun, J. D. McKenna, J. M. Murray, E. M. Ostap, and Y. E. Goldman, “Parallax: high accuracy three-dimensional single molecule tracking using split images,” Nano Lett.9(7), 2676–2682 (2009).
[CrossRef] [PubMed]

Goodwin, P. M.

N. P. Wells, G. A. Lessard, P. M. Goodwin, M. E. Phipps, P. J. Cutler, D. S. Lidke, B. S. Wilson, and J. H. Werner, “Time-resolved three-dimensional molecular tracking in live cells,” Nano Lett.10(11), 4732–4737 (2010).
[CrossRef] [PubMed]

G. A. Lessard, P. M. Goodwin, and J. H. Werner, “Three-dimensional tracking of individual quantum dots,ˮ,” Appl. Phys. Lett.91(22), 224106 (2007).
[CrossRef]

Gratton, E.

Y. Katayama, O. Burkacky, M. Meyer, C. Bräuchle, E. Gratton, and D. C. Lamb, “Real-time nanomicroscopy via three-dimensional single-particle tracking,” ChemPhysChem10(14), 2458–2464 (2009).
[CrossRef] [PubMed]

Guilford, W. H.

M. K. Cheezum, W. F. Walker, and W. H. Guilford, “Quantitative comparison of algorithms for tracking single fluorescent particles,” Biophys. J.81(4), 2378–2388 (2001).
[CrossRef] [PubMed]

Häberlein, H.

R. Veith, T. Sorkalla, E. Baumgart, J. Anzt, H. Häberlein, S. Tyagi, J. P. Siebrasse, and U. Kubitscheck, “Balbiani ring mRNPs diffuse through and bind to clusters of large intranuclear molecular structures,” Biophys. J.99(8), 2676–2685 (2010).
[CrossRef] [PubMed]

Hain, S.

Hell, S. W.

Hellmann, M.

D. Ernst, M. Hellmann, J. Kohler, and M. Weiss, “Fractional Brownian motion in crowded fluids,ˮ,” Soft Matter8(18), 4886–4889 (2012).
[CrossRef]

Holtzer, L.

L. Holtzer, T. Meckel, and T. Schmidt, “Nanometric three-dimensional tracking of individual quantum dots in cells,ˮ,” Appl. Phys. Lett.90(5), 053902 (2007).
[CrossRef]

Huisken, J.

Izeddin, I.

Jacobson, K.

M. J. Saxton and K. Jacobson, “Single-particle tracking: applications to membrane dynamics,” Annu. Rev. Biophys. Biomol. Struct.26(1), 373–399 (1997).
[CrossRef] [PubMed]

Juette, M. F.

M. F. Juette and J. Bewersdorf, “Three-dimensional tracking of single fluorescent particles with submillisecond temporal resolution,” Nano Lett.10(11), 4657–4663 (2010).
[CrossRef] [PubMed]

M. J. Mlodzianoski, M. F. Juette, G. L. Beane, and J. Bewersdorf, “Experimental characterization of 3D localization techniques for particle-tracking and super-resolution microscopy,” Opt. Express17(10), 8264–8277 (2009).
[CrossRef] [PubMed]

Kaminski, T.

J. P. Siebrasse, T. Kaminski, and U. Kubitscheck, “Nuclear export of single native mRNA molecules observed by light sheet fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.109(24), 9426–9431 (2012).
[CrossRef] [PubMed]

J. G. Ritter, J.-H. Spille, T. Kaminski, and U. Kubitscheck, “A cylindrical zoom lens unit for adjustable optical sectioning in light sheet microscopy,” Biomed. Opt. Express2(1), 185–193 (2011).
[CrossRef] [PubMed]

Kao, H. P.

H. P. Kao and A. S. Verkman, “Tracking of single fluorescent particles in three dimensions: use of cylindrical optics to encode particle position,” Biophys. J.67(3), 1291–1300 (1994).
[CrossRef] [PubMed]

Katayama, Y.

Y. Katayama, O. Burkacky, M. Meyer, C. Bräuchle, E. Gratton, and D. C. Lamb, “Real-time nanomicroscopy via three-dimensional single-particle tracking,” ChemPhysChem10(14), 2458–2464 (2009).
[CrossRef] [PubMed]

Kim, K.-A.

N. Arhel, A. Genovesio, K.-A. Kim, S. Miko, E. Perret, J.-C. Olivo-Marin, S. Shorte, and P. Charneau, “Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes,” Nat. Methods3(10), 817–824 (2006).
[CrossRef] [PubMed]

Kohler, J.

D. Ernst, M. Hellmann, J. Kohler, and M. Weiss, “Fractional Brownian motion in crowded fluids,ˮ,” Soft Matter8(18), 4886–4889 (2012).
[CrossRef]

Köhler, J.

Kubitscheck, U.

J. P. Siebrasse, T. Kaminski, and U. Kubitscheck, “Nuclear export of single native mRNA molecules observed by light sheet fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.109(24), 9426–9431 (2012).
[CrossRef] [PubMed]

J. G. Ritter, J.-H. Spille, T. Kaminski, and U. Kubitscheck, “A cylindrical zoom lens unit for adjustable optical sectioning in light sheet microscopy,” Biomed. Opt. Express2(1), 185–193 (2011).
[CrossRef] [PubMed]

R. Veith, T. Sorkalla, E. Baumgart, J. Anzt, H. Häberlein, S. Tyagi, J. P. Siebrasse, and U. Kubitscheck, “Balbiani ring mRNPs diffuse through and bind to clusters of large intranuclear molecular structures,” Biophys. J.99(8), 2676–2685 (2010).
[CrossRef] [PubMed]

J. G. Ritter, R. Veith, A. Veenendaal, J. P. Siebrasse, and U. Kubitscheck, “Light sheet microscopy for single molecule tracking in living tissue,” PLoS ONE5(7), e11639 (2010).
[CrossRef] [PubMed]

J. P. Siebrasse, R. Veith, A. Dobay, H. Leonhardt, B. Daneholt, and U. Kubitscheck, “Discontinuous movement of mRNP particles in nucleoplasmic regions devoid of chromatin,” Proc. Natl. Acad. Sci. U.S.A.105(51), 20291–20296 (2008).
[CrossRef] [PubMed]

J. G. Ritter, R. Veith, J.-P. Siebrasse, and U. Kubitscheck, “High-contrast single-particle tracking by selective focal plane illumination microscopy,” Opt. Express16(10), 7142–7152 (2008).
[CrossRef] [PubMed]

Lamb, D. C.

Y. Katayama, O. Burkacky, M. Meyer, C. Bräuchle, E. Gratton, and D. C. Lamb, “Real-time nanomicroscopy via three-dimensional single-particle tracking,” ChemPhysChem10(14), 2458–2464 (2009).
[CrossRef] [PubMed]

Lavagnino, Z.

F. Cella Zanacchi, Z. Lavagnino, M. Perrone Donnorso, A. Del Bue, L. Furia, M. Faretta, and A. Diaspro, “Live-cell 3D super-resolution imaging in thick biological samples,” Nat. Methods8(12), 1047–1049 (2011).
[CrossRef] [PubMed]

Leonhardt, H.

J. P. Siebrasse, R. Veith, A. Dobay, H. Leonhardt, B. Daneholt, and U. Kubitscheck, “Discontinuous movement of mRNP particles in nucleoplasmic regions devoid of chromatin,” Proc. Natl. Acad. Sci. U.S.A.105(51), 20291–20296 (2008).
[CrossRef] [PubMed]

Lessard, G. A.

N. P. Wells, G. A. Lessard, P. M. Goodwin, M. E. Phipps, P. J. Cutler, D. S. Lidke, B. S. Wilson, and J. H. Werner, “Time-resolved three-dimensional molecular tracking in live cells,” Nano Lett.10(11), 4732–4737 (2010).
[CrossRef] [PubMed]

G. A. Lessard, P. M. Goodwin, and J. H. Werner, “Three-dimensional tracking of individual quantum dots,ˮ,” Appl. Phys. Lett.91(22), 224106 (2007).
[CrossRef]

Lew, M. D.

M. A. Thompson, M. D. Lew, M. Badieirostami, and W. E. Moerner, “Localizing and tracking single nanoscale emitters in three dimensions with high spatiotemporal resolution using a double-helix point spread function,” Nano Lett.10(1), 211–218 (2010).
[CrossRef] [PubMed]

Liddle, J. A.

M. D. McMahon, A. J. Berglund, P. Carmichael, J. J. McClelland, and J. A. Liddle, “3D particle trajectories observed by orthogonal tracking microscopy,” ACS Nano3(3), 609–614 (2009).
[CrossRef] [PubMed]

Lidke, D. S.

N. P. Wells, G. A. Lessard, P. M. Goodwin, M. E. Phipps, P. J. Cutler, D. S. Lidke, B. S. Wilson, and J. H. Werner, “Time-resolved three-dimensional molecular tracking in live cells,” Nano Lett.10(11), 4732–4737 (2010).
[CrossRef] [PubMed]

Mabuchi, H.

K. McHale, A. J. Berglund, and H. Mabuchi, “Quantum dot photon statistics measured by three-dimensional particle tracking,” Nano Lett.7(11), 3535–3539 (2007).
[CrossRef] [PubMed]

McClelland, J. J.

M. D. McMahon, A. J. Berglund, P. Carmichael, J. J. McClelland, and J. A. Liddle, “3D particle trajectories observed by orthogonal tracking microscopy,” ACS Nano3(3), 609–614 (2009).
[CrossRef] [PubMed]

McHale, K.

K. McHale, A. J. Berglund, and H. Mabuchi, “Quantum dot photon statistics measured by three-dimensional particle tracking,” Nano Lett.7(11), 3535–3539 (2007).
[CrossRef] [PubMed]

McKenna, J. D.

Y. Sun, J. D. McKenna, J. M. Murray, E. M. Ostap, and Y. E. Goldman, “Parallax: high accuracy three-dimensional single molecule tracking using split images,” Nano Lett.9(7), 2676–2682 (2009).
[CrossRef] [PubMed]

McMahon, M. D.

M. D. McMahon, A. J. Berglund, P. Carmichael, J. J. McClelland, and J. A. Liddle, “3D particle trajectories observed by orthogonal tracking microscopy,” ACS Nano3(3), 609–614 (2009).
[CrossRef] [PubMed]

Meckel, T.

L. Holtzer, T. Meckel, and T. Schmidt, “Nanometric three-dimensional tracking of individual quantum dots in cells,ˮ,” Appl. Phys. Lett.90(5), 053902 (2007).
[CrossRef]

Meyer, M.

Y. Katayama, O. Burkacky, M. Meyer, C. Bräuchle, E. Gratton, and D. C. Lamb, “Real-time nanomicroscopy via three-dimensional single-particle tracking,” ChemPhysChem10(14), 2458–2464 (2009).
[CrossRef] [PubMed]

Miko, S.

N. Arhel, A. Genovesio, K.-A. Kim, S. Miko, E. Perret, J.-C. Olivo-Marin, S. Shorte, and P. Charneau, “Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes,” Nat. Methods3(10), 817–824 (2006).
[CrossRef] [PubMed]

Mlodzianoski, M. J.

Moerner, W. E.

M. A. Thompson, J. M. Casolari, M. Badieirostami, P. O. Brown, and W. E. Moerner, “Three-dimensional tracking of single mRNA particles in Saccharomyces cerevisiae using a double-helix point spread function,” Proc. Natl. Acad. Sci. U.S.A.107(42), 17864–17871 (2010).
[CrossRef] [PubMed]

M. A. Thompson, M. D. Lew, M. Badieirostami, and W. E. Moerner, “Localizing and tracking single nanoscale emitters in three dimensions with high spatiotemporal resolution using a double-helix point spread function,” Nano Lett.10(1), 211–218 (2010).
[CrossRef] [PubMed]

Murray, J. M.

Y. Sun, J. D. McKenna, J. M. Murray, E. M. Ostap, and Y. E. Goldman, “Parallax: high accuracy three-dimensional single molecule tracking using split images,” Nano Lett.9(7), 2676–2682 (2009).
[CrossRef] [PubMed]

Neyts, K.

H. Deschout, K. Neyts, and K. Braeckmans, “The influence of movement on the localization precision of sub-resolution particles in fluorescence microscopy,” J Biophotonics5(1), 97–109 (2012).
[CrossRef] [PubMed]

Ober, R. J.

S. Ram, P. Prabhat, J. Chao, E. S. Ward, and R. J. Ober, “High accuracy 3D quantum dot tracking with multifocal plane microscopy for the study of fast intracellular dynamics in live cells,” Biophys. J.95(12), 6025–6043 (2008).
[CrossRef] [PubMed]

Olivo-Marin, J.-C.

N. Arhel, A. Genovesio, K.-A. Kim, S. Miko, E. Perret, J.-C. Olivo-Marin, S. Shorte, and P. Charneau, “Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes,” Nat. Methods3(10), 817–824 (2006).
[CrossRef] [PubMed]

Ostap, E. M.

Y. Sun, J. D. McKenna, J. M. Murray, E. M. Ostap, and Y. E. Goldman, “Parallax: high accuracy three-dimensional single molecule tracking using split images,” Nano Lett.9(7), 2676–2682 (2009).
[CrossRef] [PubMed]

Perret, E.

N. Arhel, A. Genovesio, K.-A. Kim, S. Miko, E. Perret, J.-C. Olivo-Marin, S. Shorte, and P. Charneau, “Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes,” Nat. Methods3(10), 817–824 (2006).
[CrossRef] [PubMed]

Perrone Donnorso, M.

F. Cella Zanacchi, Z. Lavagnino, M. Perrone Donnorso, A. Del Bue, L. Furia, M. Faretta, and A. Diaspro, “Live-cell 3D super-resolution imaging in thick biological samples,” Nat. Methods8(12), 1047–1049 (2011).
[CrossRef] [PubMed]

Phipps, M. E.

N. P. Wells, G. A. Lessard, P. M. Goodwin, M. E. Phipps, P. J. Cutler, D. S. Lidke, B. S. Wilson, and J. H. Werner, “Time-resolved three-dimensional molecular tracking in live cells,” Nano Lett.10(11), 4732–4737 (2010).
[CrossRef] [PubMed]

Prabhat, P.

S. Ram, P. Prabhat, J. Chao, E. S. Ward, and R. J. Ober, “High accuracy 3D quantum dot tracking with multifocal plane microscopy for the study of fast intracellular dynamics in live cells,” Biophys. J.95(12), 6025–6043 (2008).
[CrossRef] [PubMed]

Ram, S.

S. Ram, P. Prabhat, J. Chao, E. S. Ward, and R. J. Ober, “High accuracy 3D quantum dot tracking with multifocal plane microscopy for the study of fast intracellular dynamics in live cells,” Biophys. J.95(12), 6025–6043 (2008).
[CrossRef] [PubMed]

Ritter, J. G.

Saxton, M. J.

M. J. Saxton and K. Jacobson, “Single-particle tracking: applications to membrane dynamics,” Annu. Rev. Biophys. Biomol. Struct.26(1), 373–399 (1997).
[CrossRef] [PubMed]

Schmidt, T.

L. Holtzer, T. Meckel, and T. Schmidt, “Nanometric three-dimensional tracking of individual quantum dots in cells,ˮ,” Appl. Phys. Lett.90(5), 053902 (2007).
[CrossRef]

Schönle, A.

Selvin, P. R.

E. Toprak, H. Balci, B. H. Blehm, and P. R. Selvin, “Three-dimensional particle tracking via bifocal imaging,” Nano Lett.7(7), 2043–2045 (2007).
[CrossRef] [PubMed]

Shaevitz, J. W.

Shorte, S.

N. Arhel, A. Genovesio, K.-A. Kim, S. Miko, E. Perret, J.-C. Olivo-Marin, S. Shorte, and P. Charneau, “Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes,” Nat. Methods3(10), 817–824 (2006).
[CrossRef] [PubMed]

Siebrasse, J. P.

J. P. Siebrasse, T. Kaminski, and U. Kubitscheck, “Nuclear export of single native mRNA molecules observed by light sheet fluorescence microscopy,” Proc. Natl. Acad. Sci. U.S.A.109(24), 9426–9431 (2012).
[CrossRef] [PubMed]

R. Veith, T. Sorkalla, E. Baumgart, J. Anzt, H. Häberlein, S. Tyagi, J. P. Siebrasse, and U. Kubitscheck, “Balbiani ring mRNPs diffuse through and bind to clusters of large intranuclear molecular structures,” Biophys. J.99(8), 2676–2685 (2010).
[CrossRef] [PubMed]

J. G. Ritter, R. Veith, A. Veenendaal, J. P. Siebrasse, and U. Kubitscheck, “Light sheet microscopy for single molecule tracking in living tissue,” PLoS ONE5(7), e11639 (2010).
[CrossRef] [PubMed]

J. P. Siebrasse, R. Veith, A. Dobay, H. Leonhardt, B. Daneholt, and U. Kubitscheck, “Discontinuous movement of mRNP particles in nucleoplasmic regions devoid of chromatin,” Proc. Natl. Acad. Sci. U.S.A.105(51), 20291–20296 (2008).
[CrossRef] [PubMed]

Siebrasse, J.-P.

Sorkalla, T.

R. Veith, T. Sorkalla, E. Baumgart, J. Anzt, H. Häberlein, S. Tyagi, J. P. Siebrasse, and U. Kubitscheck, “Balbiani ring mRNPs diffuse through and bind to clusters of large intranuclear molecular structures,” Biophys. J.99(8), 2676–2685 (2010).
[CrossRef] [PubMed]

Spille, J.-H.

Stainier, D. Y.

Sun, Y.

Y. Sun, J. D. McKenna, J. M. Murray, E. M. Ostap, and Y. E. Goldman, “Parallax: high accuracy three-dimensional single molecule tracking using split images,” Nano Lett.9(7), 2676–2682 (2009).
[CrossRef] [PubMed]

Thompson, M. A.

M. A. Thompson, M. D. Lew, M. Badieirostami, and W. E. Moerner, “Localizing and tracking single nanoscale emitters in three dimensions with high spatiotemporal resolution using a double-helix point spread function,” Nano Lett.10(1), 211–218 (2010).
[CrossRef] [PubMed]

M. A. Thompson, J. M. Casolari, M. Badieirostami, P. O. Brown, and W. E. Moerner, “Three-dimensional tracking of single mRNA particles in Saccharomyces cerevisiae using a double-helix point spread function,” Proc. Natl. Acad. Sci. U.S.A.107(42), 17864–17871 (2010).
[CrossRef] [PubMed]

Toprak, E.

E. Toprak, H. Balci, B. H. Blehm, and P. R. Selvin, “Three-dimensional particle tracking via bifocal imaging,” Nano Lett.7(7), 2043–2045 (2007).
[CrossRef] [PubMed]

Tyagi, S.

R. Veith, T. Sorkalla, E. Baumgart, J. Anzt, H. Häberlein, S. Tyagi, J. P. Siebrasse, and U. Kubitscheck, “Balbiani ring mRNPs diffuse through and bind to clusters of large intranuclear molecular structures,” Biophys. J.99(8), 2676–2685 (2010).
[CrossRef] [PubMed]

Veenendaal, A.

J. G. Ritter, R. Veith, A. Veenendaal, J. P. Siebrasse, and U. Kubitscheck, “Light sheet microscopy for single molecule tracking in living tissue,” PLoS ONE5(7), e11639 (2010).
[CrossRef] [PubMed]

Veith, R.

J. G. Ritter, R. Veith, A. Veenendaal, J. P. Siebrasse, and U. Kubitscheck, “Light sheet microscopy for single molecule tracking in living tissue,” PLoS ONE5(7), e11639 (2010).
[CrossRef] [PubMed]

R. Veith, T. Sorkalla, E. Baumgart, J. Anzt, H. Häberlein, S. Tyagi, J. P. Siebrasse, and U. Kubitscheck, “Balbiani ring mRNPs diffuse through and bind to clusters of large intranuclear molecular structures,” Biophys. J.99(8), 2676–2685 (2010).
[CrossRef] [PubMed]

J. G. Ritter, R. Veith, J.-P. Siebrasse, and U. Kubitscheck, “High-contrast single-particle tracking by selective focal plane illumination microscopy,” Opt. Express16(10), 7142–7152 (2008).
[CrossRef] [PubMed]

J. P. Siebrasse, R. Veith, A. Dobay, H. Leonhardt, B. Daneholt, and U. Kubitscheck, “Discontinuous movement of mRNP particles in nucleoplasmic regions devoid of chromatin,” Proc. Natl. Acad. Sci. U.S.A.105(51), 20291–20296 (2008).
[CrossRef] [PubMed]

Verkman, A. S.

H. P. Kao and A. S. Verkman, “Tracking of single fluorescent particles in three dimensions: use of cylindrical optics to encode particle position,” Biophys. J.67(3), 1291–1300 (1994).
[CrossRef] [PubMed]

von Middendorff, C.

Walker, W. F.

M. K. Cheezum, W. F. Walker, and W. H. Guilford, “Quantitative comparison of algorithms for tracking single fluorescent particles,” Biophys. J.81(4), 2378–2388 (2001).
[CrossRef] [PubMed]

Ward, E. S.

S. Ram, P. Prabhat, J. Chao, E. S. Ward, and R. J. Ober, “High accuracy 3D quantum dot tracking with multifocal plane microscopy for the study of fast intracellular dynamics in live cells,” Biophys. J.95(12), 6025–6043 (2008).
[CrossRef] [PubMed]

Weiss, M.

D. Ernst, M. Hellmann, J. Kohler, and M. Weiss, “Fractional Brownian motion in crowded fluids,ˮ,” Soft Matter8(18), 4886–4889 (2012).
[CrossRef]

Wells, N. P.

N. P. Wells, G. A. Lessard, P. M. Goodwin, M. E. Phipps, P. J. Cutler, D. S. Lidke, B. S. Wilson, and J. H. Werner, “Time-resolved three-dimensional molecular tracking in live cells,” Nano Lett.10(11), 4732–4737 (2010).
[CrossRef] [PubMed]

Werner, J. H.

N. P. Wells, G. A. Lessard, P. M. Goodwin, M. E. Phipps, P. J. Cutler, D. S. Lidke, B. S. Wilson, and J. H. Werner, “Time-resolved three-dimensional molecular tracking in live cells,” Nano Lett.10(11), 4732–4737 (2010).
[CrossRef] [PubMed]

G. A. Lessard, P. M. Goodwin, and J. H. Werner, “Three-dimensional tracking of individual quantum dots,ˮ,” Appl. Phys. Lett.91(22), 224106 (2007).
[CrossRef]

Wilson, B. S.

N. P. Wells, G. A. Lessard, P. M. Goodwin, M. E. Phipps, P. J. Cutler, D. S. Lidke, B. S. Wilson, and J. H. Werner, “Time-resolved three-dimensional molecular tracking in live cells,” Nano Lett.10(11), 4732–4737 (2010).
[CrossRef] [PubMed]

ACS Nano

M. D. McMahon, A. J. Berglund, P. Carmichael, J. J. McClelland, and J. A. Liddle, “3D particle trajectories observed by orthogonal tracking microscopy,” ACS Nano3(3), 609–614 (2009).
[CrossRef] [PubMed]

Annu. Rev. Biophys. Biomol. Struct.

M. J. Saxton and K. Jacobson, “Single-particle tracking: applications to membrane dynamics,” Annu. Rev. Biophys. Biomol. Struct.26(1), 373–399 (1997).
[CrossRef] [PubMed]

Appl. Opt.

Appl. Phys. Lett.

G. A. Lessard, P. M. Goodwin, and J. H. Werner, “Three-dimensional tracking of individual quantum dots,ˮ,” Appl. Phys. Lett.91(22), 224106 (2007).
[CrossRef]

L. Holtzer, T. Meckel, and T. Schmidt, “Nanometric three-dimensional tracking of individual quantum dots in cells,ˮ,” Appl. Phys. Lett.90(5), 053902 (2007).
[CrossRef]

Biomed. Opt. Express

Biophys. J.

R. Veith, T. Sorkalla, E. Baumgart, J. Anzt, H. Häberlein, S. Tyagi, J. P. Siebrasse, and U. Kubitscheck, “Balbiani ring mRNPs diffuse through and bind to clusters of large intranuclear molecular structures,” Biophys. J.99(8), 2676–2685 (2010).
[CrossRef] [PubMed]

M. K. Cheezum, W. F. Walker, and W. H. Guilford, “Quantitative comparison of algorithms for tracking single fluorescent particles,” Biophys. J.81(4), 2378–2388 (2001).
[CrossRef] [PubMed]

S. Ram, P. Prabhat, J. Chao, E. S. Ward, and R. J. Ober, “High accuracy 3D quantum dot tracking with multifocal plane microscopy for the study of fast intracellular dynamics in live cells,” Biophys. J.95(12), 6025–6043 (2008).
[CrossRef] [PubMed]

H. P. Kao and A. S. Verkman, “Tracking of single fluorescent particles in three dimensions: use of cylindrical optics to encode particle position,” Biophys. J.67(3), 1291–1300 (1994).
[CrossRef] [PubMed]

ChemPhysChem

Y. Katayama, O. Burkacky, M. Meyer, C. Bräuchle, E. Gratton, and D. C. Lamb, “Real-time nanomicroscopy via three-dimensional single-particle tracking,” ChemPhysChem10(14), 2458–2464 (2009).
[CrossRef] [PubMed]

J Biophotonics

H. Deschout, K. Neyts, and K. Braeckmans, “The influence of movement on the localization precision of sub-resolution particles in fluorescence microscopy,” J Biophotonics5(1), 97–109 (2012).
[CrossRef] [PubMed]

J. Opt. Soc. Am. A

Nano Lett.

E. Toprak, H. Balci, B. H. Blehm, and P. R. Selvin, “Three-dimensional particle tracking via bifocal imaging,” Nano Lett.7(7), 2043–2045 (2007).
[CrossRef] [PubMed]

Y. Sun, J. D. McKenna, J. M. Murray, E. M. Ostap, and Y. E. Goldman, “Parallax: high accuracy three-dimensional single molecule tracking using split images,” Nano Lett.9(7), 2676–2682 (2009).
[CrossRef] [PubMed]

M. F. Juette and J. Bewersdorf, “Three-dimensional tracking of single fluorescent particles with submillisecond temporal resolution,” Nano Lett.10(11), 4657–4663 (2010).
[CrossRef] [PubMed]

K. McHale, A. J. Berglund, and H. Mabuchi, “Quantum dot photon statistics measured by three-dimensional particle tracking,” Nano Lett.7(11), 3535–3539 (2007).
[CrossRef] [PubMed]

N. P. Wells, G. A. Lessard, P. M. Goodwin, M. E. Phipps, P. J. Cutler, D. S. Lidke, B. S. Wilson, and J. H. Werner, “Time-resolved three-dimensional molecular tracking in live cells,” Nano Lett.10(11), 4732–4737 (2010).
[CrossRef] [PubMed]

M. A. Thompson, M. D. Lew, M. Badieirostami, and W. E. Moerner, “Localizing and tracking single nanoscale emitters in three dimensions with high spatiotemporal resolution using a double-helix point spread function,” Nano Lett.10(1), 211–218 (2010).
[CrossRef] [PubMed]

Nat. Methods

F. Cella Zanacchi, Z. Lavagnino, M. Perrone Donnorso, A. Del Bue, L. Furia, M. Faretta, and A. Diaspro, “Live-cell 3D super-resolution imaging in thick biological samples,” Nat. Methods8(12), 1047–1049 (2011).
[CrossRef] [PubMed]

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Supplementary Material (3)

» Media 1: MPG (3538 KB)     
» Media 2: MPG (5842 KB)     
» Media 3: MPG (4618 KB)     

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Figures (4)

Fig. 1
Fig. 1

(a) Schematic representation of the instrument. Illumination light was focused into the sample chamber by a 10x apochromat objective to form a thin light sheet in the focal plane (z0). The sample could be positioned coarsely by a 3-axes translation stage (X, Y, Z) and more accurately by an additional z piezo stage (Zp). Fluorescence was collected by a high NA 40x water-immersion objective, cleared up by notch and bandpass filters (NF, BF) and detected with a camera (EMCCD). A cylindrical lens (C) was used to shape the PSF for 3D localization. Fast image analysis was used to determine this information in real-time and feed it back to the z piezo stage in an active feedback loop to keep a particle near the focal plane. (b) Detailed view of the specimen within the sample chamber. Fluorescent particles located below (1) or above (3) the focal plane resulted in an elliptical PSF as observed on the camera chip (z0).

Fig. 2
Fig. 2

(a) x-y-cross sections of the astigmatic PSF at different axial positions. Scale bar 1 µm. (b) The spread of the astigmatic PSF, wx (squares) and wy (circles), was determined by least squares fitting of an elliptical Gaussian and the results were fit by 3rd order polynomials (solid lines) yielding unambiguous information on the axial localization over an axial range > 1 µm. (c) For a smaller range (> 0.6 µm), a linear approximation of the ellipticity wx / wy represented a fast estimator for axial localization, which required only the slope as a parameter, since per definition the intercept ϵ(z = 0) = 1.

Fig. 3
Fig. 3

(a) An immobilized fluorescent bead was moved stepwise through the focal plane to determine the experimental localization precision. The bead position was determined in 100 frames per step (full black line), the mean position was marked in light gray. (b) Histograms of all localizations relative to the mean value in one plane were fit by a Gaussian function (gray) with the standard deviation σ quantifying the localization precision: σx = 4.5 nm, σy = 4.4 nm and σz = 19 nm. (c) A 100 nm nanoparticle diluted in PBS was dynamically tracked for almost 13 s (Supplementary Media 1). The x (light gray), y (dark gray) and z (black) coordinates of the bead were used to reconstruct the 3D trajectory (d, black line) with about 800 positions spanning 10 µm x 10 µm x 60 µm. 2D projections were shown to exhibit the 3D coordinates of the trajectory.

Fig. 4
Fig. 4

(a) 3D rendering of the nuclear envelope of a C. tentans salivary gland cell nucleus (green) and real-time trajectories of 20 nm fluorescent beads. Each trajectory was shown in a different color (Supplementary Media 2). (b) Individual trajectories span more than 4500 frames and (10 µm)3 (Supplementary Media 3). View point rotated and x-y-axes inverted with respect to (a) for better visibility. (c) 3D mean square displacement analysis of 5 trajectories with more than 1000 positions. (d) Jump distance and mean square displacement (inset) analysis for the trajectory displayed in (b), yielding an unimodal JD histogram and similar MSD plots in x, y and z.

Equations (2)

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( x c , y c )= | i x 0 |5; | j y 0 |5 ( I(i,j) I bg ) ×( i,j ) / | i x 0 |5; | j y 0 |5 ( I(i,j) I bg ) ,
( w x , w y ) 2 = | i x c |5; | j y c |5 ( I(i,j) I bg ) × ( i,j ) 2 / | i x c |5; | j y c |5 ( I(i,j) I bg ) .

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