Two-photon MINFLUX with doubled localization precision

eLight - 2022
Kaixuan Zhao1, Xinzhu Xu1, Weili Ren1, Dayong Jin2, Peng Xi2
1Department of Biomedical Engineering, College of Future Technology, Peking University, Beijing 100871, China
2UTS-SUStech Joint Research Centre for Biomedical Materials & Devices, Department of Biomedical Engineering, College of Engineering, Southern University of Science and Technology, Shenzhen, Guangdong, 518055, China

Tóm tắt

AbstractAchieving localization with molecular precision has been of great interest for extending fluorescence microscopy to nanoscopy. MINFLUX pioneers this transition through point spread function (PSF) engineering, yet its performance is primarily limited by the signal-to-background ratio. Here we demonstrate theoretically that two-photon MINFLUX (2p-MINFLUX) could double its localization precision through PSF engineering by nonlinear effect. Cramér-Rao Bound (CRB) is studied as the maximum localization precision, and CRB of two-photon MINFLUX is halved compared to single-photon MINFLUX (1p-MINFLUX) in all three dimensions. Meanwhile, in order to achieve same localization precision with 1p-MINFLUX, 2p-MINFLUX requires only 1/4 of fluorescence photons. Exploiting simultaneous two-photon excitation of multiple fluorophore species, 2p-MINFLUX may have the potential for registration-free nanoscopy and multicolor tracking.

Từ khóa


Tài liệu tham khảo

S.W. Hell, J. Wichmann, Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt. Lett. 19(11), 780–782 (1994)

E. Betzig, G.H. Patterson, R. Sougrat, O.W. Lindwasser, S. Olenych, J.S. Bonifacino, M.W. Davidson, J. Lippincott-Schwartz, H.F. Hess, Imaging intracellular fluorescent proteins at nanometer resolution. Science 313(5793), 1642–1645 (2006)

M.J. Rust, M. Bates, X. Zhuang, Stochastic optical reconstruction microscopy (storm) provides sub-diffraction-limit image resolution. Nat. Methods 3(10), 793 (2006)

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

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

M.P. Backlund, Y. Shechtman, R.L. Walsworth, Fundamental precision bounds for three-dimensional optical localization microscopy with poisson statistics. Phys. Rev. Lett. 121(2), 023904 (2018)

P. Fei, J. Nie, J. Lee, Y. Ding, S. Li, H. Zhang, M. Hagiwara, T. Yu, T. Segura, C.-M. Ho, D. Zhu, T.K. Hsiai, Subvoxel light-sheet microscopy for high-resolution high-throughput volumetric imaging of large biomedical specimens. Adv. Photonics 1(1), 016002 (2019)

E. Narimanov, Resolution limit of label-free far-field microscopy. Adv. Photonics 1(5), 056003 (2019)

F. Balzarotti, Y. Eilers, K.C. Gwosch, A.H. Gynnå, V. Westphal, F.D. Stefani, J. Elf, S.W. Hell, Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes. Science 355(6325), 606–612 (2017)

Y. Eilers, H. Ta, K.C. Gwosch, F. Balzarotti, S.W. Hell, Minflux monitors rapid molecular jumps with superior spatiotemporal resolution. Proc. Natl. Acad. Sci. 115(24), 6117–6122 (2018)

J.K. Pape, T. Stephan, F. Balzarotti, R. Büchner, F. Lange, D. Riedel, S. Jakobs, S.W. Hell, Multicolor 3d minflux nanoscopy of mitochondrial micos proteins. Proc. Natl. Acad. Sci. 117(34), 20607–20614 (2020)

K.C. Gwosch, J.K. Pape, F. Balzarotti, P. Hoess, J. Ellenberg, J. Ries, S.W. Hell, Minflux nanoscopy delivers 3d multicolor nanometer resolution in cells. Nat. Methods 17(2), 217–224 (2020)

L.A. Masullo, F. Steiner, J. Zähringer, L.F. Lopez, J. Bohlen, L. Richter, F. Cole, P. Tinnefeld, F.D. Stefani, Pulsed interleaved minflux. Nano Lett. 21(1), 840–846 (2021)

S.W. Hell, E.H. Stelzer, S. Lindek, C. Cremer, Confocal microscopy with an increased detection aperture: type-b 4pi confocal microscopy. Opt. Lett. 19(3), 222–224 (1994)

M.G. Gustafsson, D. Agard, J. Sedat, I5m: 3d widefield light microscopy with better than 100nm axial resolution. J. Microsc. 195(1), 10–16 (1999)

J. Bewersdorf, R. Schmidt, S.W. Hell, Comparison of i5m and 4pi-microscopy. J. Microsc. 222(2), 105–117 (2006)

J. Xiao, T. Ha, Flipping nanoscopy on its head. Science 355(6325), 582–584 (2017)

W.L. Peticolas, J.P. Goldsborough, K. Rieckhoff, Double photon excitation in organic crystals. Phys. Rev. Lett. 10(2), 43 (1963)

C.J.R. Sheppard, M. Gu, Image formation in two-photon fluorescence microscopy. Optik (Stuttgart) 86(3), 104–106 (1990)

W. Denk, J.H. Strickler, W.W. Webb, Two-photon laser scanning fluorescence microscopy. Science 248(4951), 73–76 (1990)

W. Denk, D.W. Piston, W.W. Webb, Multi-photon molecular excitation in laser-scanning microscopy. In: Handbook of Biological Confocal Microscopy (Springer, Boston, MA 2006), pp. 535–549

F. Bestvater, E. Spiess, G. Stobrawa, M. Hacker, T. Feurer, T. Porwol, U. Berchner-Pfannschmidt, C. Wotzlaw, H. Acker, Two-photon fluorescence absorption and emission spectra of dyes relevant for cell imaging. J. Microsc. 208(2), 108–115 (2002)

J. Mütze, V. Iyer, J.J. Macklin, J. Colonell, B. Karsh, Z. Petrášek, P. Schwille, L.L. Looger, L.D. Lavis, T.D. Harris, Excitation spectra and brightness optimization of two-photon excited probes. Biophys. J. 102(4), 934–944 (2012)

M.G.M. Velasco, E.S. Allgeyer, P. Yuan, J. Grutzendler, J. Bewersdorf, Absolute two-photon excitation spectra of red and far-red fluorescent probes. Opt. Lett. 40(21), 4915–4918 (2015)

K. Zhao, X. Xu, W. Ren, D. Jin, P. Xi, Code for 2p-MINFLUX simulation. github (2021), https://github.com/kunzhao1220/2pMINFLUX

D. Kobat, M.E. Durst, N. Nishimura, A.W. Wong, C.B. Schaffer, C. Xu, Deep tissue multiphoton microscopy using longer wavelength excitation. Opt. Express 17(16), 13354–13364 (2009)

P. Schwille, U. Haupts, S. Maiti, W.W. Webb, Molecular dynamics in living cells observed by fluorescence correlation spectroscopy with one- and two-photon excitation. Biophys. J. 77(4), 2251–2265 (1999)

K.G. Heinze, A. Koltermann, P. Schwille, Simultaneous two-photon excitation of distinct labels for dual-color fluorescence crosscorrelation analysis. Proc. Natl. Acad. Sci. 97(19), 10377–10382 (2000)

P.S. Dittrich, P. Schwille, Photobleaching and stabilization of fluorophores used for single-molecule analysis with one- and two-photon excitation. Appl. Phys. B 73(8), 829–837 (2001)

J. Mütze, Z. Petrášek, P. Schwille, Independence of maximum single molecule fluorescence count rate on the temporal and spectral laser pulse width in two-photon fcs. J. Fluoresc. 17(6), 805–810 (2007)