An alternative to MINFLUX that enables nanometer resolution in a confocal microscope

Luciano A. Masullo; Alan M. Szalai; Lucía F. Lopez; Mauricio Pilo-Pais; Guillermo P. Acuna; Fernando D. Stefani

doi:10.1038/s41377-022-00896-4

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An alternative to MINFLUX that enables nanometer resolution in a confocal microscope

Original Articles

- An alternative to MINFLUX that enables nanometer resolution in a confocal microscope
- An alternative to MINFLUX that enables nanometer resolution in a confocal microscope
- Light: Science & Applications 2022年11卷第9期页码：2077-2085
- Affiliations：
  
  1.Centro de Investigaciones en Bionanociencias (CIBION), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2390, C1425FQD Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
  2.Departamento de Física, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Güiraldes 2620, C1428EHA Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina
  3.Department of Physics, University of Fribourg, Chemin du Musée 3, Fribourg CH-1700, Switzerland
- Author bio：
  
  Fernando D. Stefani (fernando.stefani@df.uba.ar)
- Funds：
- DOI：10.1038/s41377-022-00896-4
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Luciano A. Masullo, Alan M. Szalai, Lucía F. Lopez, 等. An alternative to MINFLUX that enables nanometer resolution in a confocal microscope[J]. Light: Science & Applications, 2022,11(9):2077-2085.

Masullo, L. A. et al. An alternative to MINFLUX that enables nanometer resolution in a confocal microscope. Light: Science & Applications, 11, 2077-2085 (2022).
Luciano A. Masullo, Alan M. Szalai, Lucía F. Lopez, 等. An alternative to MINFLUX that enables nanometer resolution in a confocal microscope[J]. Light: Science & Applications, 2022,11(9):2077-2085. DOI： 10.1038/s41377-022-00896-4.

Masullo, L. A. et al. An alternative to MINFLUX that enables nanometer resolution in a confocal microscope. Light: Science & Applications, 11, 2077-2085 (2022). DOI： 10.1038/s41377-022-00896-4.

摘要

Abstract

Localization of single fluorescent emitters is key for physicochemical and biophysical measurements at the nanoscale and beyond ensemble averaging. Examples include single-molecule tracking and super-resolution imaging by single-molecule localization microscopy. Among the numerous localization methods available, MINFLUX outstands for achieving a ~10-fold improvement in resolution over wide-field camera-based approaches, reaching the molecular scale at moderate photon counts. Widespread application of MINFLUX and related methods has been hindered by the technical complexity of the setups. Here, we present RASTMIN, a single-molecule localization method based on raster scanning a light pattern comprising a minimum of intensity. RASTMIN delivers ~1–2 nm localization precision with usual fluorophores and is easily implementable on a standard confocal microscope with few modifications. We demonstrate the performance of RASTMIN in localization of single molecules and super-resolution imaging of DNA origami structures.

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references

M. Orrit , J. Bernard . Single pentacene molecules detected by fluorescence excitation in a p-terphenyl crystal . Phys. Rev. Lett. , 1990 . 65 2716 -2719 . DOI:10.1103/PhysRevLett.65.2716http://doi.org/10.1103/PhysRevLett.65.2716 .

L. Kador , D. E. Horne , W. E. Moerner . Optical detection and probing of single dopant molecules of pentacene in a p-terphenyl host crystal by means of absorption spectroscopy . J. Phys. Chem. , 1990 . 94 1237 -1248 . DOI:10.1021/j100367a011http://doi.org/10.1021/j100367a011 .

F. Kulzer , M. Orrit . Single-molecule optics . Annu. Rev. Phys. Chem. , 2004 . 55 585 -611 . DOI:10.1146/annurev.physchem.54.011002.103816http://doi.org/10.1146/annurev.physchem.54.011002.103816 .

M. Orrit , T. Ha , V. Sandoghdar . Single-molecule optical spectroscopy . Chem. Soc. Rev. , 2014 . 43 973 DOI:10.1039/c4cs90001dhttp://doi.org/10.1039/c4cs90001d .

D. Brinks , 等 . Ultrafast dynamics of single molecules . Chem. Soc. Rev. , 2014 . 43 2476 -2491 . DOI:10.1039/C3CS60269Ahttp://doi.org/10.1039/C3CS60269A .

P. Holzmeister , G. P. Acuna , D. Grohmann , P. Tinnefeld . Breaking the concentration limit of optical single-molecule detection . Chem. Soc. Rev. , 2014 . 43 1014 -1028 . DOI:10.1039/C3CS60207Ahttp://doi.org/10.1039/C3CS60207A .

S. Shashkova , M. C. Leake . Single-molecule fluorescence microscopy review: shedding new light on old problems . Biosci. Rep. , 2017 . 37 1 -19 . DOI:10.1042/BSR20170031http://doi.org/10.1042/BSR20170031 .

B. Hellenkamp , 等 . Precision and accuracy of single-molecule FRET measurements—a multi-laboratory benchmark study . Nat. Methods , 2018 . 15 669 -676 . DOI:10.1038/s41592-018-0085-0http://doi.org/10.1038/s41592-018-0085-0 .

S. van de Linde , 等 . Direct stochastic optical reconstruction microscopy with standard fluorescent probes . Nat. Protoc. , 2011 . 6 991 -1009 . DOI:10.1038/nprot.2011.336http://doi.org/10.1038/nprot.2011.336 .

J. Schnitzbauer , M. T. Strauss , T. Schlichthaerle , F. Schueder , R. Jungmann . Super-resolution microscopy with DNA-PAINT . Nat. Protoc. , 2017 . 12 1198 -1228 . DOI:10.1038/nprot.2017.024http://doi.org/10.1038/nprot.2017.024 .

M. Lelek , 等 . Single-molecule localization microscopy . Nat. Rev. Methods Prim. , 2021 . 1 39 DOI:10.1038/s43586-021-00038-xhttp://doi.org/10.1038/s43586-021-00038-x .

A. M. Van Oijen , 等 . Single-molecule kinetics of λ exonuclease reveal base dependence and dynamic disorder . Science , 2003 . 301 1235 -1238 . DOI:10.1126/science.1084387http://doi.org/10.1126/science.1084387 .

A. Yildiz , 等 . Myosin V walks hand-over-hand: single fluorophore imaging with 1.5-nm localization . Science , 2003 . 300 2061 -2065. .

B. P. English , 等 . Ever-fluctuating single enzyme molecules: Michaelis-Menten equation revisited . Nat. Chem. Biol. , 2006 . 2 87 -94 . DOI:10.1038/nchembio759http://doi.org/10.1038/nchembio759 .

I. I. Cisse , 等 . Real-time dynamics of RNA polymerase II clustering in live human cells . Science , 2013 . 341 664 -667 . DOI:10.1126/science.1239053http://doi.org/10.1126/science.1239053 .

P. Kosuri , B. D. Altheimer , M. Dai , P. Yin , X. Zhuang . Rotation tracking of genome-processing enzymes using DNA origami rotors . Nature , 2019 . 572 136 -140 . DOI:10.1038/s41586-019-1397-7http://doi.org/10.1038/s41586-019-1397-7 .

R. E. Thompson , D. R. Larson , W. W. Webb . Precise nanometer localization analysis for individual fluorescent probes . Biophys. J. , 2002 . 82 2775 -2783 . DOI:10.1016/S0006-3495(02)75618-Xhttp://doi.org/10.1016/S0006-3495(02)75618-X .

K. I. Mortensen , L. S. Churchman , J. A. Spudich , H. Flyvbjerg . Optimized localization analysis for single-molecule tracking and super-resolution microscopy . Nat. Methods , 2010 . 7 377 -381 . DOI:10.1038/nmeth.1447http://doi.org/10.1038/nmeth.1447 .

L. A. Masullo , L. F. Lopez , F. D. Stefani . A common framework for single-molecule localization using sequential structured illumination . Biophys. Rep. , 2022 . 2 100036 .

J. Enderlein . Tracking of fluorescent molecules diffusing within membranes . Appl. Phys. B , 2000 . 71 773 -777 . DOI:10.1007/s003400000409http://doi.org/10.1007/s003400000409 .

V. Levi , Q. Ruan , K. Kis-Petikova , E. Gratton . Scanning FCS, a novel method for three-dimensional particle tracking . Biochem. Soc. Trans. , 2003 . 31 997 -1000 . DOI:10.1042/bst0310997http://doi.org/10.1042/bst0310997 .

Davis, L. M. et al. Four-focus single-particle position determination in a confocal microscope. SPIE Proceedings 7571: Single Molecule Spectroscopy and Imaging III, 757112 (2010).

G. A. Lessard , P. M. Goodwin , J. H. Werner . Three-dimensional tracking of individual quantum dots . Appl. Phys. Lett. , 2007 . 91 224106 DOI:10.1063/1.2819074http://doi.org/10.1063/1.2819074 .

A. Dupont , 等 . Three-dimensional single-particle tracking in live cells: news from the third dimension . N. J. Phys. , 2013 . 15 075008 DOI:10.1088/1367-2630/15/7/075008http://doi.org/10.1088/1367-2630/15/7/075008 .

J. A. Germann , L. M. Davis . Three-dimensional tracking of a single fluorescent nanoparticle using four-focus excitation in a confocal microscope . Opt. Express , 2014 . 22 5641 DOI:10.1364/OE.22.005641http://doi.org/10.1364/OE.22.005641 .

F. Balzarotti , 等 . Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes . Science , 2017 . 355 606 -612 . DOI:10.1126/science.aak9913http://doi.org/10.1126/science.aak9913 .

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. USA , 2018 . 115 6117 -6122 . DOI:10.1073/pnas.1801672115http://doi.org/10.1073/pnas.1801672115 .

L. A. Masullo , 等 . Pulsed Interleaved MINFLUX . Nano Lett. , 2021 . 21 840 -846 . DOI:10.1021/acs.nanolett.0c04600http://doi.org/10.1021/acs.nanolett.0c04600 .

J. K. Pape , 等 . Multicolor 3D MINFLUX nanoscopy of mitochondrial MICOS proteins . Proc. Natl Acad. Sci. USA , 2020 . 117 20607 -20614 . DOI:10.1073/pnas.2009364117http://doi.org/10.1073/pnas.2009364117 .

K. C. Gwosch , 等 . MINFLUX nanoscopy delivers 3D multicolor nanometer resolution in cells . Nat. Methods , 2020 . 17 217 -224 . DOI:10.1038/s41592-019-0688-0http://doi.org/10.1038/s41592-019-0688-0 .

L. Gu , 等 . Molecular resolution imaging by repetitive optical selective exposure . Nat. Methods , 2019 . 16 1114 -1118 . DOI:10.1038/s41592-019-0544-2http://doi.org/10.1038/s41592-019-0544-2 .

L. Reymond , 等 . SIMPLE: Structured illumination based point localization estimator with enhanced precision . Optics Express , 2019 . 27 24578 -24590 . DOI:10.1364/OE.27.024578http://doi.org/10.1364/OE.27.024578 .

J. Cnossen , 等 . Localization microscopy at doubled precision with patterned illumination . Nat. Methods , 2020 . 17 59 -63 . DOI:10.1038/s41592-019-0657-7http://doi.org/10.1038/s41592-019-0657-7 .

P. Jouchet , 等 . Nanometric axial localization of single fluorescent molecules with modulated excitation . Nature Photonics , 2021 . 15 297 -304 . DOI:10.1038/s41566-020-00749-9http://doi.org/10.1038/s41566-020-00749-9 .

K. Zhao , X. Xu , W. Ren , D. Jin , P. Xi . Two-photon MINFLUX with doubled localization precision . eLight , 2022 . 2 5 DOI:10.1186/s43593-021-00011-xhttp://doi.org/10.1186/s43593-021-00011-x .

L. A. Masullo , F. D. Stefani . Multiphoton single-molecule localization by sequential excitation with light minima . Light Sci. Appl. , 2022 . 11 70 DOI:10.1038/s41377-022-00763-2http://doi.org/10.1038/s41377-022-00763-2 .

J. C. Thiele , 等 . Confocal fluorescence-lifetime single-molecule localization microscopy . ACS Nano , 2020 . 14 14190 -14200 . DOI:10.1021/acsnano.0c07322http://doi.org/10.1021/acsnano.0c07322 .

R. Schmidt , 等 . MINFLUX nanometer-scale 3D imaging and microsecond-range tracking on a common fluorescence microscope . Nat. Commun. , 2021 . 12 1478 DOI:10.1038/s41467-021-21652-zhttp://doi.org/10.1038/s41467-021-21652-z .

L. A. Masullo , A. M. Szalai , L. F. Lopez , F. D. Stefani . Fluorescence nanoscopy at the sub-10 nm scale . Biophys. Rev. , 2021 . 13 1101 -1112 . DOI:10.1007/s12551-021-00864-zhttp://doi.org/10.1007/s12551-021-00864-z .

L. A. Masullo , 等 . Enhanced photon collection enables four dimensional fluorescence nanoscopy of living systems . Nat. Commun. , 2018 . 9 3281 DOI:10.1038/s41467-018-05799-whttp://doi.org/10.1038/s41467-018-05799-w .

M. Castello , 等 . A robust and versatile platform for image scanning microscopy enabling super-resolution FLIM . Nat. Methods , 2019 . 16 175 -178 . DOI:10.1038/s41592-018-0291-9http://doi.org/10.1038/s41592-018-0291-9 .

A. M. Szalai , 等 . Three-dimensional total-internal reflection fluorescence nanoscopy with nanometric axial resolution by photometric localization of single molecules . Nat. Commun. , 2021 . 12 517 DOI:10.1038/s41467-020-20863-0http://doi.org/10.1038/s41467-020-20863-0 .

F. Caprile , L. A. Masullo , F. D. Stefani . PyFocus – a Python package for vectorial calculations of focused optical fields under realistic conditions. Application to toroidal foci . Comput. Phys. Commun. , 2022 . 275 108315 DOI:10.1016/j.cpc.2022.108315http://doi.org/10.1016/j.cpc.2022.108315 .

A. Ghosh , 等 . Graphene-based metal-induced energy transfer for sub-nanometre optical localization . Nat. Photonics , 2019 . 13 860 -865 . DOI:10.1038/s41566-019-0510-7http://doi.org/10.1038/s41566-019-0510-7 .

A. Ghosh , A. I. Chizhik , N. Karedla , J. Enderlein . Graphene- and metal-induced energy transfer for single-molecule imaging and live-cell nanoscopy with (sub)-nanometer axial resolution . Nat. Protoc. , 2021 . 16 3695 -3715 . DOI:10.1038/s41596-021-00558-6http://doi.org/10.1038/s41596-021-00558-6 .

I. Kaminska , 等 . Distance dependence of single-molecule energy transfer to graphene measured with DNA origami nanopositioners . Nano Lett. , 2019 . 19 4257 -4262 . DOI:10.1021/acs.nanolett.9b00172http://doi.org/10.1021/acs.nanolett.9b00172 .

I. Kamińska , 等 . Graphene energy transfer for single-molecule biophysics, biosensing, and super-resolution microscopy . Adv. Mater. , 2021 . 33 2101099 DOI:10.1002/adma.202101099http://doi.org/10.1002/adma.202101099 .

S. M. Douglas , 等 . Rapid prototyping of 3D DNA-origami shapes with caDNAno . Nucleic Acids Res. , 2009 . 37 5001 -5006 . DOI:10.1093/nar/gkp436http://doi.org/10.1093/nar/gkp436 .

E.-M. Roller , C. Argyropoulos , A. Högele , T. Liedl , M. Pilo-Pais . Plasmon–exciton coupling using DNA templates . Nano Lett. , 2016 . 16 5962 -5966 . DOI:10.1021/acs.nanolett.6b03015http://doi.org/10.1021/acs.nanolett.6b03015 .

N. G. Bastús , J. Comenge , V. Puntes . Kinetically controlled seeded growth synthesis of citrate-stabilized gold nanoparticles of up to 200 nm: size focusing versus Ostwald ripening . Langmuir , 2011 . 27 11098 -11105 . DOI:10.1021/la201938uhttp://doi.org/10.1021/la201938u .

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