Abstract

Nanoscale particle tracking in three dimensions is crucial to directly observe dynamics of molecules and nanoparticles in living cells. Here we present a three-dimensional particle tracking method based on temporally focused two-photon excitation. Multiple particles are imaged at 30 frames/s in volume up to 180 × 180 × 100 µm3. The spatial localization precision can reach 50 nm. We demonstrate its capability of tracking fast swimming microbes at speed of ~200 µm/s. Two-photon dual-color tracking is achieved by simultaneously exciting two kinds of fluorescent beads at 800 nm to demonstrate its potential in molecular interaction studies. Our method provides a simple wide-field fluorescence imaging approach for deep multiple-particle tracking.

© 2016 Optical Society of America

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References

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2015 (6)

Y. Shechtman, L. E. Weiss, A. S. Backer, S. J. Sahl, and W. E. Moerner, “Precise Three-Dimensional Scan-Free Multiple-Particle Tracking over Large Axial Ranges with Tetrapod Point Spread Functions,” Nano Lett. 15(6), 4194–4199 (2015).
[Crossref] [PubMed]

E. P. Perillo, Y.-L. Liu, K. Huynh, C. Liu, C.-K. Chou, M.-C. Hung, H.-C. Yeh, and A. K. Dunn, “Deep and high-resolution three-dimensional tracking of single particles using nonlinear and multiplexed illumination,” Nat. Commun. 6, 7874 (2015).
[Crossref] [PubMed]

A. Huhle, D. Klaue, H. Brutzer, P. Daldrop, S. Joo, O. Otto, U. F. Keyser, and R. Seidel, “Camera-based three-dimensional real-time particle tracking at kHz rates and Ångström accuracy,” Nat. Commun. 6, 5885 (2015).
[Crossref] [PubMed]

K. M. Taute, S. Gude, S. J. Tans, and T. S. Shimizu, “High-throughput 3D tracking of bacteria on a standard phase contrast microscope,” Nat. Commun. 6, 8776 (2015).
[Crossref] [PubMed]

K. Son, D. R. Brumley, and R. Stocker, “Live from under the lens: exploring microbial motility with dynamic imaging and microfluidics,” Nat. Rev. Microbiol. 13(12), 761–775 (2015).
[Crossref] [PubMed]

R. Spesyvtsev, H. A. Rendall, and K. Dholakia, “Wide-field three-dimensional optical imaging using temporal focusing for holographically trapped microparticles,” Opt. Lett. 40(21), 4847–4850 (2015).
[Crossref] [PubMed]

2014 (8)

C.-H. Lien, C.-Y. Lin, S.-J. Chen, and F.-C. Chien, “Dynamic particle tracking via temporal focusing multiphoton microscopy with astigmatism imaging,” Opt. Express 22(22), 27290–27299 (2014).
[Crossref] [PubMed]

A. Small and S. Stahlheber, “Fluorophore localization algorithms for super-resolution microscopy,” Nat. Methods 11(3), 267–279 (2014).
[Crossref] [PubMed]

S. Jia, J. C. Vaughan, and X. Zhuang, “Isotropic three-dimensional super-resolution imaging with a self-bending point spread function,” Nat. Photonics 8(4), 302–306 (2014).
[Crossref] [PubMed]

Y. Shechtman, S. J. Sahl, A. S. Backer, and W. E. Moerner, “Optimal Point Spread Function Design for 3D Imaging,” Phys. Rev. Lett. 113(13), 133902 (2014).
[Crossref] [PubMed]

H. Deschout, F. Cella Zanacchi, M. Mlodzianoski, A. Diaspro, J. Bewersdorf, S. T. Hess, and K. Braeckmans, “Precisely and accurately localizing single emitters in fluorescence microscopy,” Nat. Methods 11(3), 253–266 (2014).
[Crossref] [PubMed]

A. Kusumi, T. A. Tsunoyama, K. M. Hirosawa, R. S. Kasai, and T. K. Fujiwara, “Tracking single molecules at work in living cells,” Nat. Chem. Biol. 10(7), 524–532 (2014).
[Crossref] [PubMed]

H. Y. Park, H. Lim, Y. J. Yoon, A. Follenzi, C. Nwokafor, M. Lopez-Jones, X. Meng, and R. H. Singer, “Visualization of Dynamics of Single Endogenous mRNA Labeled in Live Mouse,” Science 343(6169), 422–424 (2014).
[Crossref] [PubMed]

W. Wang and N. Tao, “Detection, counting, and imaging of single nanoparticles,” Anal. Chem. 86(1), 2–14 (2014).
[Crossref] [PubMed]

2013 (3)

2012 (2)

2011 (3)

2010 (1)

E. Papagiakoumou, F. Anselmi, A. Bègue, V. de Sars, J. Glückstad, E. Y. Isacoff, and V. Emiliani, “Scanless two-photon excitation of channelrhodopsin-2,” Nat. Methods 7(10), 848–854 (2010).
[Crossref] [PubMed]

2009 (4)

J. Yu, C. Wu, S. P. Sahu, L. P. Fernando, C. Szymanski, and J. McNeill, “Nanoscale 3D Tracking with Conjugated Polymer Nanoparticles,” J. Am. Chem. Soc. 131(51), 18410–18414 (2009).
[Crossref] [PubMed]

R. Lindken, M. Rossi, S. Grosse, and J. Westerweel, “Micro-Particle Image Velocimetry (microPIV): recent developments, applications, and guidelines,” Lab Chip 9(17), 2551–2567 (2009).
[Crossref] [PubMed]

Y. Katayama, O. Burkacky, M. Meyer, C. Bräuchle, E. Gratton, and D. C. Lamb, “Real-Time Nanomicroscopy Via Three-Dimensional Single-Particle Tracking,” ChemPhysChem 10(14), 2458–2464 (2009).
[Crossref] [PubMed]

S. R. P. Pavani, M. A. Thompson, J. S. Biteen, S. J. Lord, N. Liu, R. J. Twieg, R. Piestun, and W. E. Moerner, “Three-dimensional, single-molecule fluorescence imaging beyond the diffraction limit by using a double-helix point spread function,” Proc. Natl. Acad. Sci. U.S.A. 106(9), 2995–2999 (2009).
[Crossref] [PubMed]

2008 (3)

B. Huang, W. Wang, M. Bates, and X. Zhuang, “Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy,” Science 319(5864), 810–813 (2008).
[Crossref] [PubMed]

M. E. Durst, G. Zhu, and C. Xu, “Simultaneous spatial and temporal focusing in nonlinear microscopy,” Opt. Commun. 281(7), 1796–1805 (2008).
[Crossref] [PubMed]

A. Vaziri, J. Tang, H. Shroff, and C. V. Shank, “Multilayer three-dimensional super resolution imaging of thick biological samples,” Proc. Natl. Acad. Sci. U.S.A. 105(51), 20221–20226 (2008).
[Crossref] [PubMed]

2007 (2)

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]

C. Li, W. Wagner, M. Ciocca, and W. S. Warren, “Multiphoton femtosecond phase-coherent two-dimensional electronic spectroscopy,” J. Chem. Phys. 126(16), 164307 (2007).
[Crossref] [PubMed]

2006 (2)

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging Intracellular Fluorescent Proteins at Nanometer Resolution,” Science 313(5793), 1642–1645 (2006).
[Crossref] [PubMed]

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006).
[Crossref] [PubMed]

2005 (5)

V. Levi, Q. Ruan, and E. Gratton, “3-D particle tracking in a two-photon microscope: application to the study of molecular dynamics in cells,” Biophys. J. 88(4), 2919–2928 (2005).
[Crossref] [PubMed]

M. Wu, J. W. Roberts, and M. Buckley, “Three-dimensional fluorescent particle tracking at micron-scale using a single camera,” Exp. Fluids 38(4), 461–465 (2005).
[Crossref]

V. Levi, Q. Ruan, M. Plutz, A. S. Belmont, and E. Gratton, “Chromatin Dynamics in Interphase Cells Revealed by Tracking in a Two-Photon Excitation Microscope,” Biophys. J. 89(6), 4275–4285 (2005).
[Crossref] [PubMed]

D. Oron, E. Tal, and Y. Silberberg, “Scanningless depth-resolved microscopy,” Opt. Express 13(5), 1468–1476 (2005).
[Crossref] [PubMed]

G. Zhu, J. van Howe, M. Durst, W. Zipfel, and C. Xu, “Simultaneous spatial and temporal focusing of femtosecond pulses,” Opt. Express 13(6), 2153–2159 (2005).
[Crossref] [PubMed]

2003 (2)

M. Speidel, A. Jonás, and E.-L. Florin, “Three-dimensional tracking of fluorescent nanoparticles with subnanometer precision by use of off-focus imaging,” Opt. Lett. 28(2), 69–71 (2003).
[Crossref] [PubMed]

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[Crossref] [PubMed]

2000 (2)

A. M. Weiner, “Femtosecond pulse shaping using spatial light modulators,” Rev. Sci. Instrum. 71(5), 1929–1960 (2000).
[Crossref]

J. Boenigk and H. Arndt, “Particle handling during interception feeding by four species of heterotrophic nanoflagellates,” J. Eukaryot. Microbiol. 47(4), 350–358 (2000).
[Crossref] [PubMed]

1998 (1)

I. M. Peters, B. G. de Grooth, J. M. Schins, C. G. Figdor, and J. Greve, “Three dimensional single-particle tracking with nanometer resolution,” Rev. Sci. Instrum. 69(7), 2762–2766 (1998).
[Crossref]

1997 (1)

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]

1990 (1)

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248(4951), 73–76 (1990).
[Crossref] [PubMed]

1989 (1)

1988 (1)

T. Fenchel, “Marine Plankton Food Chains,” Annu. Rev. Ecol. Syst. 19(1), 19–38 (1988).
[Crossref]

Ammar, D. A.

Anselmi, F.

E. Papagiakoumou, F. Anselmi, A. Bègue, V. de Sars, J. Glückstad, E. Y. Isacoff, and V. Emiliani, “Scanless two-photon excitation of channelrhodopsin-2,” Nat. Methods 7(10), 848–854 (2010).
[Crossref] [PubMed]

Arndt, H.

J. Boenigk and H. Arndt, “Particle handling during interception feeding by four species of heterotrophic nanoflagellates,” J. Eukaryot. Microbiol. 47(4), 350–358 (2000).
[Crossref] [PubMed]

Aubé, B.

Aumayr, K.

T. Schrödel, R. Prevedel, K. Aumayr, M. Zimmer, and A. Vaziri, “Brain-wide 3D imaging of neuronal activity in Caenorhabditis elegans with sculpted light,” Nat. Methods 10(10), 1013–1020 (2013).
[Crossref] [PubMed]

Backer, A. S.

Y. Shechtman, L. E. Weiss, A. S. Backer, S. J. Sahl, and W. E. Moerner, “Precise Three-Dimensional Scan-Free Multiple-Particle Tracking over Large Axial Ranges with Tetrapod Point Spread Functions,” Nano Lett. 15(6), 4194–4199 (2015).
[Crossref] [PubMed]

Y. Shechtman, S. J. Sahl, A. S. Backer, and W. E. Moerner, “Optimal Point Spread Function Design for 3D Imaging,” Phys. Rev. Lett. 113(13), 133902 (2014).
[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]

Bates, M.

B. Huang, W. Wang, M. Bates, and X. Zhuang, “Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy,” Science 319(5864), 810–813 (2008).
[Crossref] [PubMed]

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006).
[Crossref] [PubMed]

Bègue, A.

E. Papagiakoumou, F. Anselmi, A. Bègue, V. de Sars, J. Glückstad, E. Y. Isacoff, and V. Emiliani, “Scanless two-photon excitation of channelrhodopsin-2,” Nat. Methods 7(10), 848–854 (2010).
[Crossref] [PubMed]

Belmont, A. S.

V. Levi, Q. Ruan, M. Plutz, A. S. Belmont, and E. Gratton, “Chromatin Dynamics in Interphase Cells Revealed by Tracking in a Two-Photon Excitation Microscope,” Biophys. J. 89(6), 4275–4285 (2005).
[Crossref] [PubMed]

Betzig, E.

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging Intracellular Fluorescent Proteins at Nanometer Resolution,” Science 313(5793), 1642–1645 (2006).
[Crossref] [PubMed]

Bewersdorf, J.

H. Deschout, F. Cella Zanacchi, M. Mlodzianoski, A. Diaspro, J. Bewersdorf, S. T. Hess, and K. Braeckmans, “Precisely and accurately localizing single emitters in fluorescence microscopy,” Nat. Methods 11(3), 253–266 (2014).
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T. Schrödel, R. Prevedel, K. Aumayr, M. Zimmer, and A. Vaziri, “Brain-wide 3D imaging of neuronal activity in Caenorhabditis elegans with sculpted light,” Nat. Methods 10(10), 1013–1020 (2013).
[Crossref] [PubMed]

Rendall, H. A.

Roberts, J. W.

M. Wu, J. W. Roberts, and M. Buckley, “Three-dimensional fluorescent particle tracking at micron-scale using a single camera,” Exp. Fluids 38(4), 461–465 (2005).
[Crossref]

Rossi, M.

R. Lindken, M. Rossi, S. Grosse, and J. Westerweel, “Micro-Particle Image Velocimetry (microPIV): recent developments, applications, and guidelines,” Lab Chip 9(17), 2551–2567 (2009).
[Crossref] [PubMed]

Ruan, Q.

V. Levi, Q. Ruan, and E. Gratton, “3-D particle tracking in a two-photon microscope: application to the study of molecular dynamics in cells,” Biophys. J. 88(4), 2919–2928 (2005).
[Crossref] [PubMed]

V. Levi, Q. Ruan, M. Plutz, A. S. Belmont, and E. Gratton, “Chromatin Dynamics in Interphase Cells Revealed by Tracking in a Two-Photon Excitation Microscope,” Biophys. J. 89(6), 4275–4285 (2005).
[Crossref] [PubMed]

Rust, M. J.

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006).
[Crossref] [PubMed]

Ruthardt, N.

N. Ruthardt, D. C. Lamb, and C. Bräuchle, “Single-particle Tracking as a Quantitative Microscopy-based Approach to Unravel Cell Entry Mechanisms of Viruses and Pharmaceutical Nanoparticles,” Mol. Ther. 19(7), 1199–1211 (2011).
[Crossref] [PubMed]

Sahl, S. J.

Y. Shechtman, L. E. Weiss, A. S. Backer, S. J. Sahl, and W. E. Moerner, “Precise Three-Dimensional Scan-Free Multiple-Particle Tracking over Large Axial Ranges with Tetrapod Point Spread Functions,” Nano Lett. 15(6), 4194–4199 (2015).
[Crossref] [PubMed]

Y. Shechtman, S. J. Sahl, A. S. Backer, and W. E. Moerner, “Optimal Point Spread Function Design for 3D Imaging,” Phys. Rev. Lett. 113(13), 133902 (2014).
[Crossref] [PubMed]

Sahu, S. P.

J. Yu, C. Wu, S. P. Sahu, L. P. Fernando, C. Szymanski, and J. McNeill, “Nanoscale 3D Tracking with Conjugated Polymer Nanoparticles,” J. Am. Chem. Soc. 131(51), 18410–18414 (2009).
[Crossref] [PubMed]

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]

Schins, J. M.

I. M. Peters, B. G. de Grooth, J. M. Schins, C. G. Figdor, and J. Greve, “Three dimensional single-particle tracking with nanometer resolution,” Rev. Sci. Instrum. 69(7), 2762–2766 (1998).
[Crossref]

Schrödel, T.

T. Schrödel, R. Prevedel, K. Aumayr, M. Zimmer, and A. Vaziri, “Brain-wide 3D imaging of neuronal activity in Caenorhabditis elegans with sculpted light,” Nat. Methods 10(10), 1013–1020 (2013).
[Crossref] [PubMed]

Seidel, R.

A. Huhle, D. Klaue, H. Brutzer, P. Daldrop, S. Joo, O. Otto, U. F. Keyser, and R. Seidel, “Camera-based three-dimensional real-time particle tracking at kHz rates and Ångström accuracy,” Nat. Commun. 6, 5885 (2015).
[Crossref] [PubMed]

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]

Shank, C. V.

A. Vaziri, J. Tang, H. Shroff, and C. V. Shank, “Multilayer three-dimensional super resolution imaging of thick biological samples,” Proc. Natl. Acad. Sci. U.S.A. 105(51), 20221–20226 (2008).
[Crossref] [PubMed]

Shechtman, Y.

Y. Shechtman, L. E. Weiss, A. S. Backer, S. J. Sahl, and W. E. Moerner, “Precise Three-Dimensional Scan-Free Multiple-Particle Tracking over Large Axial Ranges with Tetrapod Point Spread Functions,” Nano Lett. 15(6), 4194–4199 (2015).
[Crossref] [PubMed]

Y. Shechtman, S. J. Sahl, A. S. Backer, and W. E. Moerner, “Optimal Point Spread Function Design for 3D Imaging,” Phys. Rev. Lett. 113(13), 133902 (2014).
[Crossref] [PubMed]

Sheppard, C. J. R.

Shimizu, T. S.

K. M. Taute, S. Gude, S. J. Tans, and T. S. Shimizu, “High-throughput 3D tracking of bacteria on a standard phase contrast microscope,” Nat. Commun. 6, 8776 (2015).
[Crossref] [PubMed]

Shroff, H.

A. Vaziri, J. Tang, H. Shroff, and C. V. Shank, “Multilayer three-dimensional super resolution imaging of thick biological samples,” Proc. Natl. Acad. Sci. U.S.A. 105(51), 20221–20226 (2008).
[Crossref] [PubMed]

Silberberg, Y.

Singer, R. H.

H. Y. Park, H. Lim, Y. J. Yoon, A. Follenzi, C. Nwokafor, M. Lopez-Jones, X. Meng, and R. H. Singer, “Visualization of Dynamics of Single Endogenous mRNA Labeled in Live Mouse,” Science 343(6169), 422–424 (2014).
[Crossref] [PubMed]

Small, A.

A. Small and S. Stahlheber, “Fluorophore localization algorithms for super-resolution microscopy,” Nat. Methods 11(3), 267–279 (2014).
[Crossref] [PubMed]

So, P. T. C.

Son, K.

K. Son, D. R. Brumley, and R. Stocker, “Live from under the lens: exploring microbial motility with dynamic imaging and microfluidics,” Nat. Rev. Microbiol. 13(12), 761–775 (2015).
[Crossref] [PubMed]

Sougrat, R.

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging Intracellular Fluorescent Proteins at Nanometer Resolution,” Science 313(5793), 1642–1645 (2006).
[Crossref] [PubMed]

Speidel, M.

Spesyvtsev, R.

Spille, J.-H.

Squier, J.

Stahlheber, S.

A. Small and S. Stahlheber, “Fluorophore localization algorithms for super-resolution microscopy,” Nat. Methods 11(3), 267–279 (2014).
[Crossref] [PubMed]

Stocker, R.

K. Son, D. R. Brumley, and R. Stocker, “Live from under the lens: exploring microbial motility with dynamic imaging and microfluidics,” Nat. Rev. Microbiol. 13(12), 761–775 (2015).
[Crossref] [PubMed]

Straub, A.

Strickler, J. H.

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248(4951), 73–76 (1990).
[Crossref] [PubMed]

Szymanski, C.

J. Yu, C. Wu, S. P. Sahu, L. P. Fernando, C. Szymanski, and J. McNeill, “Nanoscale 3D Tracking with Conjugated Polymer Nanoparticles,” J. Am. Chem. Soc. 131(51), 18410–18414 (2009).
[Crossref] [PubMed]

Tal, E.

Tang, J.

A. Vaziri, J. Tang, H. Shroff, and C. V. Shank, “Multilayer three-dimensional super resolution imaging of thick biological samples,” Proc. Natl. Acad. Sci. U.S.A. 105(51), 20221–20226 (2008).
[Crossref] [PubMed]

Tans, S. J.

K. M. Taute, S. Gude, S. J. Tans, and T. S. Shimizu, “High-throughput 3D tracking of bacteria on a standard phase contrast microscope,” Nat. Commun. 6, 8776 (2015).
[Crossref] [PubMed]

Tao, N.

W. Wang and N. Tao, “Detection, counting, and imaging of single nanoparticles,” Anal. Chem. 86(1), 2–14 (2014).
[Crossref] [PubMed]

Taute, K. M.

K. M. Taute, S. Gude, S. J. Tans, and T. S. Shimizu, “High-throughput 3D tracking of bacteria on a standard phase contrast microscope,” Nat. Commun. 6, 8776 (2015).
[Crossref] [PubMed]

Therrien, O. D.

Thompson, M. A.

S. R. P. Pavani, M. A. Thompson, J. S. Biteen, S. J. Lord, N. Liu, R. J. Twieg, R. Piestun, and W. E. Moerner, “Three-dimensional, single-molecule fluorescence imaging beyond the diffraction limit by using a double-helix point spread function,” Proc. Natl. Acad. Sci. U.S.A. 106(9), 2995–2999 (2009).
[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]

Tsunoyama, T. A.

A. Kusumi, T. A. Tsunoyama, K. M. Hirosawa, R. S. Kasai, and T. K. Fujiwara, “Tracking single molecules at work in living cells,” Nat. Chem. Biol. 10(7), 524–532 (2014).
[Crossref] [PubMed]

Twieg, R. J.

S. R. P. Pavani, M. A. Thompson, J. S. Biteen, S. J. Lord, N. Liu, R. J. Twieg, R. Piestun, and W. E. Moerner, “Three-dimensional, single-molecule fluorescence imaging beyond the diffraction limit by using a double-helix point spread function,” Proc. Natl. Acad. Sci. U.S.A. 106(9), 2995–2999 (2009).
[Crossref] [PubMed]

van Howe, J.

Vaughan, J. C.

S. Jia, J. C. Vaughan, and X. Zhuang, “Isotropic three-dimensional super-resolution imaging with a self-bending point spread function,” Nat. Photonics 8(4), 302–306 (2014).
[Crossref] [PubMed]

Vaziri, A.

T. Schrödel, R. Prevedel, K. Aumayr, M. Zimmer, and A. Vaziri, “Brain-wide 3D imaging of neuronal activity in Caenorhabditis elegans with sculpted light,” Nat. Methods 10(10), 1013–1020 (2013).
[Crossref] [PubMed]

A. Vaziri, J. Tang, H. Shroff, and C. V. Shank, “Multilayer three-dimensional super resolution imaging of thick biological samples,” Proc. Natl. Acad. Sci. U.S.A. 105(51), 20221–20226 (2008).
[Crossref] [PubMed]

Vitek, D.

Wagner, W.

C. Li, W. Wagner, M. Ciocca, and W. S. Warren, “Multiphoton femtosecond phase-coherent two-dimensional electronic spectroscopy,” J. Chem. Phys. 126(16), 164307 (2007).
[Crossref] [PubMed]

Wang, W.

W. Wang and N. Tao, “Detection, counting, and imaging of single nanoparticles,” Anal. Chem. 86(1), 2–14 (2014).
[Crossref] [PubMed]

B. Huang, W. Wang, M. Bates, and X. Zhuang, “Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy,” Science 319(5864), 810–813 (2008).
[Crossref] [PubMed]

Warren, W. S.

C. Li, W. Wagner, M. Ciocca, and W. S. Warren, “Multiphoton femtosecond phase-coherent two-dimensional electronic spectroscopy,” J. Chem. Phys. 126(16), 164307 (2007).
[Crossref] [PubMed]

Webb, W. W.

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[Crossref] [PubMed]

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248(4951), 73–76 (1990).
[Crossref] [PubMed]

Weiner, A. M.

A. M. Weiner, “Femtosecond pulse shaping using spatial light modulators,” Rev. Sci. Instrum. 71(5), 1929–1960 (2000).
[Crossref]

Weiss, L. E.

Y. Shechtman, L. E. Weiss, A. S. Backer, S. J. Sahl, and W. E. Moerner, “Precise Three-Dimensional Scan-Free Multiple-Particle Tracking over Large Axial Ranges with Tetrapod Point Spread Functions,” Nano Lett. 15(6), 4194–4199 (2015).
[Crossref] [PubMed]

Westerweel, J.

R. Lindken, M. Rossi, S. Grosse, and J. Westerweel, “Micro-Particle Image Velocimetry (microPIV): recent developments, applications, and guidelines,” Lab Chip 9(17), 2551–2567 (2009).
[Crossref] [PubMed]

Williams, R. M.

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[Crossref] [PubMed]

Wu, C.

J. Yu, C. Wu, S. P. Sahu, L. P. Fernando, C. Szymanski, and J. McNeill, “Nanoscale 3D Tracking with Conjugated Polymer Nanoparticles,” J. Am. Chem. Soc. 131(51), 18410–18414 (2009).
[Crossref] [PubMed]

Wu, M.

M. Wu, J. W. Roberts, and M. Buckley, “Three-dimensional fluorescent particle tracking at micron-scale using a single camera,” Exp. Fluids 38(4), 461–465 (2005).
[Crossref]

Xu, C.

Yeh, H.-C.

E. P. Perillo, Y.-L. Liu, K. Huynh, C. Liu, C.-K. Chou, M.-C. Hung, H.-C. Yeh, and A. K. Dunn, “Deep and high-resolution three-dimensional tracking of single particles using nonlinear and multiplexed illumination,” Nat. Commun. 6, 7874 (2015).
[Crossref] [PubMed]

Yen, W.-C.

Yew, E. Y. S.

Yoon, Y. J.

H. Y. Park, H. Lim, Y. J. Yoon, A. Follenzi, C. Nwokafor, M. Lopez-Jones, X. Meng, and R. H. Singer, “Visualization of Dynamics of Single Endogenous mRNA Labeled in Live Mouse,” Science 343(6169), 422–424 (2014).
[Crossref] [PubMed]

Yu, J.

J. Yu, C. Wu, S. P. Sahu, L. P. Fernando, C. Szymanski, and J. McNeill, “Nanoscale 3D Tracking with Conjugated Polymer Nanoparticles,” J. Am. Chem. Soc. 131(51), 18410–18414 (2009).
[Crossref] [PubMed]

Zhu, G.

M. E. Durst, G. Zhu, and C. Xu, “Simultaneous spatial and temporal focusing in nonlinear microscopy,” Opt. Commun. 281(7), 1796–1805 (2008).
[Crossref] [PubMed]

G. Zhu, J. van Howe, M. Durst, W. Zipfel, and C. Xu, “Simultaneous spatial and temporal focusing of femtosecond pulses,” Opt. Express 13(6), 2153–2159 (2005).
[Crossref] [PubMed]

Zhuang, X.

S. Jia, J. C. Vaughan, and X. Zhuang, “Isotropic three-dimensional super-resolution imaging with a self-bending point spread function,” Nat. Photonics 8(4), 302–306 (2014).
[Crossref] [PubMed]

B. Huang, W. Wang, M. Bates, and X. Zhuang, “Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy,” Science 319(5864), 810–813 (2008).
[Crossref] [PubMed]

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006).
[Crossref] [PubMed]

Zimmer, M.

T. Schrödel, R. Prevedel, K. Aumayr, M. Zimmer, and A. Vaziri, “Brain-wide 3D imaging of neuronal activity in Caenorhabditis elegans with sculpted light,” Nat. Methods 10(10), 1013–1020 (2013).
[Crossref] [PubMed]

Zipfel, W.

Zipfel, W. R.

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[Crossref] [PubMed]

Anal. Chem. (1)

W. Wang and N. Tao, “Detection, counting, and imaging of single nanoparticles,” Anal. Chem. 86(1), 2–14 (2014).
[Crossref] [PubMed]

Annu. Rev. Biophys. Biomol. Struct. (1)

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]

Annu. Rev. Ecol. Syst. (1)

T. Fenchel, “Marine Plankton Food Chains,” Annu. Rev. Ecol. Syst. 19(1), 19–38 (1988).
[Crossref]

Biomed. Opt. Express (4)

Biophys. J. (2)

V. Levi, Q. Ruan, M. Plutz, A. S. Belmont, and E. Gratton, “Chromatin Dynamics in Interphase Cells Revealed by Tracking in a Two-Photon Excitation Microscope,” Biophys. J. 89(6), 4275–4285 (2005).
[Crossref] [PubMed]

V. Levi, Q. Ruan, and E. Gratton, “3-D particle tracking in a two-photon microscope: application to the study of molecular dynamics in cells,” Biophys. J. 88(4), 2919–2928 (2005).
[Crossref] [PubMed]

ChemPhysChem (1)

Y. Katayama, O. Burkacky, M. Meyer, C. Bräuchle, E. Gratton, and D. C. Lamb, “Real-Time Nanomicroscopy Via Three-Dimensional Single-Particle Tracking,” ChemPhysChem 10(14), 2458–2464 (2009).
[Crossref] [PubMed]

Exp. Fluids (1)

M. Wu, J. W. Roberts, and M. Buckley, “Three-dimensional fluorescent particle tracking at micron-scale using a single camera,” Exp. Fluids 38(4), 461–465 (2005).
[Crossref]

J. Am. Chem. Soc. (1)

J. Yu, C. Wu, S. P. Sahu, L. P. Fernando, C. Szymanski, and J. McNeill, “Nanoscale 3D Tracking with Conjugated Polymer Nanoparticles,” J. Am. Chem. Soc. 131(51), 18410–18414 (2009).
[Crossref] [PubMed]

J. Chem. Phys. (1)

C. Li, W. Wagner, M. Ciocca, and W. S. Warren, “Multiphoton femtosecond phase-coherent two-dimensional electronic spectroscopy,” J. Chem. Phys. 126(16), 164307 (2007).
[Crossref] [PubMed]

J. Eukaryot. Microbiol. (1)

J. Boenigk and H. Arndt, “Particle handling during interception feeding by four species of heterotrophic nanoflagellates,” J. Eukaryot. Microbiol. 47(4), 350–358 (2000).
[Crossref] [PubMed]

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

Lab Chip (1)

R. Lindken, M. Rossi, S. Grosse, and J. Westerweel, “Micro-Particle Image Velocimetry (microPIV): recent developments, applications, and guidelines,” Lab Chip 9(17), 2551–2567 (2009).
[Crossref] [PubMed]

Mol. Ther. (1)

N. Ruthardt, D. C. Lamb, and C. Bräuchle, “Single-particle Tracking as a Quantitative Microscopy-based Approach to Unravel Cell Entry Mechanisms of Viruses and Pharmaceutical Nanoparticles,” Mol. Ther. 19(7), 1199–1211 (2011).
[Crossref] [PubMed]

Nano Lett. (2)

Y. Shechtman, L. E. Weiss, A. S. Backer, S. J. Sahl, and W. E. Moerner, “Precise Three-Dimensional Scan-Free Multiple-Particle Tracking over Large Axial Ranges with Tetrapod Point Spread Functions,” Nano Lett. 15(6), 4194–4199 (2015).
[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]

Nat. Biotechnol. (1)

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[Crossref] [PubMed]

Nat. Chem. Biol. (1)

A. Kusumi, T. A. Tsunoyama, K. M. Hirosawa, R. S. Kasai, and T. K. Fujiwara, “Tracking single molecules at work in living cells,” Nat. Chem. Biol. 10(7), 524–532 (2014).
[Crossref] [PubMed]

Nat. Commun. (3)

A. Huhle, D. Klaue, H. Brutzer, P. Daldrop, S. Joo, O. Otto, U. F. Keyser, and R. Seidel, “Camera-based three-dimensional real-time particle tracking at kHz rates and Ångström accuracy,” Nat. Commun. 6, 5885 (2015).
[Crossref] [PubMed]

K. M. Taute, S. Gude, S. J. Tans, and T. S. Shimizu, “High-throughput 3D tracking of bacteria on a standard phase contrast microscope,” Nat. Commun. 6, 8776 (2015).
[Crossref] [PubMed]

E. P. Perillo, Y.-L. Liu, K. Huynh, C. Liu, C.-K. Chou, M.-C. Hung, H.-C. Yeh, and A. K. Dunn, “Deep and high-resolution three-dimensional tracking of single particles using nonlinear and multiplexed illumination,” Nat. Commun. 6, 7874 (2015).
[Crossref] [PubMed]

Nat. Methods (5)

T. Schrödel, R. Prevedel, K. Aumayr, M. Zimmer, and A. Vaziri, “Brain-wide 3D imaging of neuronal activity in Caenorhabditis elegans with sculpted light,” Nat. Methods 10(10), 1013–1020 (2013).
[Crossref] [PubMed]

H. Deschout, F. Cella Zanacchi, M. Mlodzianoski, A. Diaspro, J. Bewersdorf, S. T. Hess, and K. Braeckmans, “Precisely and accurately localizing single emitters in fluorescence microscopy,” Nat. Methods 11(3), 253–266 (2014).
[Crossref] [PubMed]

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006).
[Crossref] [PubMed]

E. Papagiakoumou, F. Anselmi, A. Bègue, V. de Sars, J. Glückstad, E. Y. Isacoff, and V. Emiliani, “Scanless two-photon excitation of channelrhodopsin-2,” Nat. Methods 7(10), 848–854 (2010).
[Crossref] [PubMed]

A. Small and S. Stahlheber, “Fluorophore localization algorithms for super-resolution microscopy,” Nat. Methods 11(3), 267–279 (2014).
[Crossref] [PubMed]

Nat. Photonics (1)

S. Jia, J. C. Vaughan, and X. Zhuang, “Isotropic three-dimensional super-resolution imaging with a self-bending point spread function,” Nat. Photonics 8(4), 302–306 (2014).
[Crossref] [PubMed]

Nat. Rev. Microbiol. (1)

K. Son, D. R. Brumley, and R. Stocker, “Live from under the lens: exploring microbial motility with dynamic imaging and microfluidics,” Nat. Rev. Microbiol. 13(12), 761–775 (2015).
[Crossref] [PubMed]

Opt. Commun. (1)

M. E. Durst, G. Zhu, and C. Xu, “Simultaneous spatial and temporal focusing in nonlinear microscopy,” Opt. Commun. 281(7), 1796–1805 (2008).
[Crossref] [PubMed]

Opt. Express (5)

Opt. Lett. (2)

Phys. Rev. Lett. (1)

Y. Shechtman, S. J. Sahl, A. S. Backer, and W. E. Moerner, “Optimal Point Spread Function Design for 3D Imaging,” Phys. Rev. Lett. 113(13), 133902 (2014).
[Crossref] [PubMed]

Proc. Natl. Acad. Sci. U.S.A. (2)

S. R. P. Pavani, M. A. Thompson, J. S. Biteen, S. J. Lord, N. Liu, R. J. Twieg, R. Piestun, and W. E. Moerner, “Three-dimensional, single-molecule fluorescence imaging beyond the diffraction limit by using a double-helix point spread function,” Proc. Natl. Acad. Sci. U.S.A. 106(9), 2995–2999 (2009).
[Crossref] [PubMed]

A. Vaziri, J. Tang, H. Shroff, and C. V. Shank, “Multilayer three-dimensional super resolution imaging of thick biological samples,” Proc. Natl. Acad. Sci. U.S.A. 105(51), 20221–20226 (2008).
[Crossref] [PubMed]

Rev. Sci. Instrum. (2)

I. M. Peters, B. G. de Grooth, J. M. Schins, C. G. Figdor, and J. Greve, “Three dimensional single-particle tracking with nanometer resolution,” Rev. Sci. Instrum. 69(7), 2762–2766 (1998).
[Crossref]

A. M. Weiner, “Femtosecond pulse shaping using spatial light modulators,” Rev. Sci. Instrum. 71(5), 1929–1960 (2000).
[Crossref]

Science (4)

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging Intracellular Fluorescent Proteins at Nanometer Resolution,” Science 313(5793), 1642–1645 (2006).
[Crossref] [PubMed]

H. Y. Park, H. Lim, Y. J. Yoon, A. Follenzi, C. Nwokafor, M. Lopez-Jones, X. Meng, and R. H. Singer, “Visualization of Dynamics of Single Endogenous mRNA Labeled in Live Mouse,” Science 343(6169), 422–424 (2014).
[Crossref] [PubMed]

B. Huang, W. Wang, M. Bates, and X. Zhuang, “Three-Dimensional Super-Resolution Imaging by Stochastic Optical Reconstruction Microscopy,” Science 319(5864), 810–813 (2008).
[Crossref] [PubMed]

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248(4951), 73–76 (1990).
[Crossref] [PubMed]

Other (1)

“iXon Ultra 897 Specification,” http://www.andor.com/scientific-cameras/ixon-emccd-camera-series/ixon-ultra-897 .

Supplementary Material (1)

NameDescription
» Visualization 1: AVI (4356 KB)      Cafeteria swimming revised video

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Figures (5)

Fig. 1
Fig. 1 Experimental setup of the temporal focusing two-photon microscope with spatial light modulation. HWP: half-wave plate, AOM: acousto-optic modulator, AFG: arbitrary function generator, DM: dichroic mirror, L: lens, BP: bandpass filter.
Fig. 2
Fig. 2 Characterization of the temporal focusing two-photon microscope using a 100 nm fluorescent nanosphere. (a) PSF of a 100 nm bead. (b) Series of fluorescent images of nanosphere at different z positions (z = −10 to 60 µm). When this nanosphere moves closer to the objective lens ( + z), defocused rings start forming. (c) Series of fluorescent images of nanosphere at different z positions (z = −10 to 60 µm) after the temporal focal plane is shifted to z = 30 µm plane with applied GVD on AOM. (d) Maximum intensity plots along axial dimension for images in (b) and (c).
Fig. 3
Fig. 3 Particle 3D position calculation based on defocused images. (a) Calculated ring radius R vs. Z relationship in large axial scan range (ΔZ = 110 µm) is nonlinear. (b) The R vs. z relationship shows linearity in short scan range (ΔZ = 11 µm). (c) Within ultrashort range (ΔZ = 180 nm) the R vs. Z relationship shows overall linearity with measurement standard deviation of 2 nm. The large kinks in this curve is due to mechanical instability. (d) Mechanical stability measurement of one stationary sphere showing its 3D position (X, Y, Z) has fluctuations with standard deviation σX, σY and σZ around 20 nm.
Fig. 4
Fig. 4 Dual-color multiple-particle imaging. (a) Overlay of both green- and red- channel images that are collected simultaneously by two camera showing 8 green spheres and 6 red spheres and 8 green spheres (bar: 20 µm). Respective (b) green channel image, and (c) red channel image. (d) 3D projection of total 14 spheres in the volume of 180 × 180 × 100 µm3.
Fig. 5
Fig. 5 Dynamic tracking of microbe Cafeteria (Cro) swimming. (a) Frame by frame images (33 ms interval) of two Cafeteria with one near stationary on the left, and the other one fast swimming on the right (see Visualization 1). (b) 3D trajectories of these two Cafeteria (black lines). (c) Instantaneous speed of two Cafeteria with color graph showing one Cafeteria is moving at high speed 100-250 µm/s, and the other one with an initial speed of over 250 µm/s and slows down to rest.

Equations (1)

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I(x,y)=A exp(k ( ( x x 0 ) 2 + ( y y 0 ) 2 r 0 ) 2 )

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