[Spectroscopie et microscopie d'une molécule unique]
Des progrès récents sur le plan de la sensibilité des détecteurs et de la conception des expériences permettent d'étudier les propriétés photophysiques de molécules uniques avec une sensibilité et des résolutions spatiale et temporelle optimales. Les données spectroscopiques et temporelles permettent d'accéder à la structure moléculaire, aux changements conformationnels, aux interactions intermoléculaires et à la nature de l'environnement d'une molécule unique. L'extension au domaine de la microscopie permet d'étudier la colocalisation de molécules uniques in vitro et in vivo avec une résolution de l'ordre du nanomètre. Ces avancées bénéficient autant à la chimie qu'à la biologie ou à la physique.
Advances in detector sensitivity and improvements in instrument design have recently provided scientists with tools to probe single molecules with light, and monitor their photophysical properties with exquisite sensitivity and spatial as well as temporal resolution. Spectroscopic and temporal information is used to explore molecular structure, conformational dynamics, local environment and intermolecular interactions of individual species. High-resolution single-molecule microscopy allows these methods to be used for in vitro or in vivo molecular colocalization with nanometer precision. The collected data have generated a wealth of new information in domains ranging from chemistry and biology to solid state physics.
Accepté le :
Publié le :
Mot clés : molécule unique, spectroscopie, fluorescence, fluorophore, microscopie confocal, microscopie optique à balayage en champ proche, durée de vie, polarisation
Xavier Michalet 1 ; Shimon Weiss 1
@article{CRPHYS_2002__3_5_619_0, author = {Xavier Michalet and Shimon Weiss}, title = {Single-molecule spectroscopy and microscopy}, journal = {Comptes Rendus. Physique}, pages = {619--644}, publisher = {Elsevier}, volume = {3}, number = {5}, year = {2002}, doi = {10.1016/S1631-0705(02)01343-9}, language = {en}, }
Xavier Michalet; Shimon Weiss. Single-molecule spectroscopy and microscopy. Comptes Rendus. Physique, Volume 3 (2002) no. 5, pp. 619-644. doi : 10.1016/S1631-0705(02)01343-9. https://comptes-rendus.academie-sciences.fr/physique/articles/10.1016/S1631-0705(02)01343-9/
[1] Examining nanoenvironments in solids on the scale of a single, isolated impurity molecule, Science, Volume 265 (1994), pp. 46-53
[2] Illuminating single molecules in condensed matter, Science, Volume 283 (1999), pp. 1670-1676
[3] Role of rare sites in single molecule spectroscopy measurements of spectral diffusion, J. Chem. Phys., Volume 114 (2001), pp. 10479-10485
[4] Single-molecule studies of heterogeneous dynamics in polymer melts near the glass transition, Science, Volume 292 (2001), pp. 255-258
[5] Fluorescence spectroscopy of single biomolecules, Science, Volume 283 (1999), pp. 1676-1683
[6] Measuring conformational dynamics of biomolecules by single molecule fluorescence spectroscopy, Nature Struct. Biol., Volume 7 (2000), pp. 724-729
[7] Single molecule nanobioscience, Trends Biochem. Sci., Volume 26 (2001), pp. 438-444
[8] Fluorescent probes and bioconjugation chemistries for single-molecule fluorescence analysis of biomolecules, J. Chem. Phys. (2002) (to be published)
[9] Single photons on demand from a single molecule at room temperature, Nature, Volume 407 (2000), pp. 491-493
[10] Single-Molecule Optical Detection, Imaging and Spectroscopy (T. Basché; W.E. Moerner; M. Orrit; U.P. Wild, eds.), VCH, Weinheim, 1997
[11] Optical detection of single molecules, Annu. Rev. Biophys. Biomol. Struct., Volume 26 (1997), pp. 567-596
[12] Single molecule spectroscopy, Annu. Rev. Phys. Chem., Volume 48 (1997), pp. 181-212
[13] Optical studies of single molecules at room temperature, Annu. Rev. Phys. Chem., Volume 49 (1998), pp. 441-480
[14] Ten years of single-molecule spectroscopy, J. Phys. Chem. A, Volume 104 (2000), pp. 1-16
[15] Fluorescence Correlation Spectroscopy. Theory and Applications (R. Rigler; E.S. Elson, eds.), Springer, 2001
[16] Single Molecule Spectroscopy (R. Rigler; M. Orrit; T. Basche, eds.), Springer-Verlag, Stockholm, 2001
[17] La fluorescence, Ann. Phys., Volume 10 (1918), pp. 133-159
[18] Development of a 500 Å spatial resolution light microscope. I. Light is efficiently transmitted through λ/16 diameter apertures, Ultramicroscopy, Volume 13 (1984), pp. 227-230
[19] Optical stethoscopy: Image recording with resolution λ/20, Appl. Phys. Lett., Volume 42 (1984), pp. 651-653
[20] Near-field optics: Microscopy, spectroscopy, and surface modification beyond the diffraction limit, Science, Volume 257 (1992), pp. 189-195
[21] Single-channel currents recorded from membrane of denervated frog muscle fibres, Nature, Volume 260 (1976), pp. 799-802
[22] Surface studies by scanning tunneling microscopy, Phys. Rev. Lett., Volume 49 (1982), pp. 57-61
[23] Atomic force microscope, Phys. Rev. Lett., Volume 56 (1986), pp. 930-933
[24] A review of tunneling microscope and atomic force microscope imaging of large biological structures – Problem and prospects, Scanning Microscopy, Volume 5 (1991), pp. 907-918
[25] Properties of biomolecules measured from atomic force microscope images: A review, J. Struct. Biol., Volume 119 (1997), pp. 99-108
[26] Optical detection and spectroscopy of single molecules in a solid, Phys. Rev. Lett., Volume 62 (1989), pp. 2535-2538
[27] Single pentacene molecules detected by fluorescence excitation in a p-terphenyl crystal, Phys. Rev. Lett., Volume 65 (1990), pp. 2716-2719
[28] Single molecules observed by near-field scanning optical microscopy, Science, Volume 262 (1993), pp. 1422-1425
[29] Optical modification of a single impurity molecule in a solid, Nature, Volume 355 (1992), pp. 335-337
[30] Near-field spectroscopy of single molecules at room temperature, Science, Volume 369 (1994), pp. 40-42
[31] Fluorescence spectroscopy and spectral diffusion of single impurity molecules in a crystal, Nature, Volume 349 (1991), pp. 225-227
[32] Detection of single fluorescent molecules, Chem. Phys. Lett., Volume 174 (1990), pp. 553-557
[33] Imaging of single molecule diffusion, Proc. Nat. Acad. Sci. USA, Volume 93 (1996), pp. 2926-2929
[34] 3D Imaging of individual ion channels in live cells at 40 nm resolution, Single Molecules, Volume 1 (2000), pp. 25-31
[35] Single-molecule imaging of signaling molecules in living cells, Single Molecules, Volume 1 (2000), pp. 159-163
[36] High photo-stability of single molecules in an organic crystal at room temperature observed by scanning confocal microscopy, Mol. Phys., Volume 95 (1998), pp. 1333-1338
[37] Terrylene in p-terphenyl: single-molecule experiments at room temperature, Chem. Phys., Volume 247 (1999), pp. 23-34
[38] Optical microscopic observation of single small molecules, Appl. Opt., Volume 15 (1976), pp. 2965-2966
[39] Detection of single molecules of phycoerythrin in hydrodynamically focused flows by laser-induced fluorescence, Anal. Chem., Volume 59 (1987), pp. 2158-2161
[40] Probing individual molecules with confocal fluorescence microscopy, Science, Volume 266 (1994), pp. 1018-1021
[41] Fluorescence correlation spectroscopy as a tool to investigate chemical reactions in solutions and on cell surfaces, Cell. Mol. Biol., Volume 44 (1998), pp. 857-879
[42] Fluorescence correlation spectroscopy and its potential for intracellular applications, Cell Biochem. Biophys., Volume 34 (2001), pp. 383-408
[43] Imaging and time-resolved spectroscopy of single molecules at an interface, Science, Volume 272 (1996), pp. 255-258
[44] Single-pair fluorescence energy transfer on freely diffusing molecules: Observation of Förster distance dependence and subpopulations, Proc. Nat. Acad. Sci. USA, Volume 96 (1999), pp. 3670-3675
[45] Confocal fluorescence lifetime imaging microscopy (flim) at the single molecule level, Single Molecules, Volume 1 (2000), pp. 215-223
[46] 3-dimensional super-resolution by spectrally selective imaging, Chem. Phys. Lett., Volume 292 (1998), pp. 183-187
[47] Ultrahigh-resolution multicolor colocalization of single fluorescent probes, Proc. Nat. Acad. Sci. USA, Volume 97 (2000), pp. 9461-9466
[48] Three-dimensional orientation measurements of symmetric single chromophores using polarization microscopy, Nature, Volume 399 (1999), pp. 126-130
[49] Fluorescence intermittency in single cadmium selenide nanocrystals, Nature, Volume 383 (1996), pp. 802-804
[50] Correlation between fluorescence intermittency and spectral diffusion in single semiconductor quantum dots, Phys. Rev. Lett., Volume 85 (2000), pp. 3301-3304
[51] Photon antibunching in the fluorescence of a single dye molecule trapped in a solid, Phys. Rev. Lett., Volume 69 (1992), pp. 1516-1519
[52] Handbook of Biological Confocal Microscopy (J.B. Pawley, ed.), Plenum Press, New York, 1995
[53] Tracking single proteins within cells, Biophys. J., Volume 79 (2000), pp. 2188-2198
[54] Imaging of single fluorescent molecules and individual ATP turnovers by single myosin molecules in aqueous solution, Nature, Volume 374 (1995), pp. 555-559
[55] A comparison of through-the-objective total internal reflection microscopy and epifluorescence microscopy for single-molecule fluorescence imaging, Single Molecules, Volume 2 (2001), pp. 191-201
[56] Ultrafast microchannel plate photomultipliers, Appl. Opt., Volume 27 (1988), pp. 1170-1178
[57] Photon counting techniques with silicon avalanche photodiodes, Appl. Opt., Volume 32 (1993), pp. 3894-3900
[58] Actively quenched single-photon avalanche diode for high-repetition rate time-gated photon counting, Rev. Sci. Instrum., Volume 67 (1996), pp. 55-61
[59] Principles of Fluorescence Spectroscopy, Plenum, New York, 1999
[60] Sub-electron read noise at MHz pixel rates, SPIE Proc., 4306, 2001, pp. 289-298
[61] Novel detectors for fluorescence lifetime imaging on the picosecond time scale, J. Fluor., Volume 7 (1997), pp. 93-98
[62] Single-molecule fluorescence – each photon counts (R. Rigler; M. Orrit; T. Basche, eds.), Book, Springer-Verlag, Stockholm, 2001
[63] On/off blinking and switching behaviour of single molecules of green fluorescent protein, Nature, Volume 388 (1997), pp. 355-358
[64] Single-molecule enzymatic dynamics, Science, Volume 282 (1998), pp. 1877-1882
[65] Autofluorescent proteins in single-molecule research: Applications to live cell imaging microscopy, Biophys. J., Volume 80 (2001), pp. 2396-2408
[66] Discrete intensity jumps and intramolcular electronic energy transfer in the spectroscopy of single conjugated polymer molecules, Science, Volume 277 (1997), pp. 1074-1077
[67] Photochemistry and fluorescence emission dynamics of single molecules in solution – B-phycoerythrin, J. Phys. Chem., Volume 100 (1996), pp. 17406-17409
[68] Fluorescence and photobleaching dynamics of single light-harvesting complexes, Proc. Nat. Acad. Sci. USA, Volume 94 (1997), pp. 10630-10635
[69] Triplet states as non-radiative traps in mulichromophoric entities: single molecule spectroscopy of an artificial and natural antenna system, Spectr. Acta A, Volume 57 (2001), pp. 2093-2107
[70] Photoluminescence spectroscopy of single CdSe nanocrystallite quantum dots, Phys. Rev. Lett., Volume 77 (1996), pp. 3873-3876
[71] Quantum jumps of single molecules at room temperature, Chem. Phys. Lett., Volume 271 (1997), pp. 1-5
[72] Time-varying triplet state lifetimes of single molecules, Phys. Rev. Lett., Volume 83 (1999), pp. 2155-2158
[73] Random telegraph signal in the photoluminescence intensity of a single quantum dot, Phys. Rev. Lett., Volume 78 (1997), pp. 1110-1113
[74] Evidence for a thermal contribution to emission intermittency in single CdSe/CdS core/shell nanocrystals, J. Chem. Phys., Volume 110 (1999), pp. 1195-1201
[75] Dual-molecule spectroscopy: Molecular rulers for the study of biological macromolecules, IEEE J. Sel. Topics Quantum Electron., Volume 2 (1996), pp. 1115-1128
[76] Probing single molecule dynamics, Science, Volume 265 (1994), pp. 361-364
[77] Single molecule dynamics studied by polarization modulation, Phys. Rev. Lett., Volume 77 (1996), pp. 3979-3982
[78] Alterations of single molecule fluorescence lifetimes in near-field optical microscopy, Science, Volume 265 (1994), pp. 364-367
[79] Influencing the angular emission of a single molecule, Phys. Rev. Lett., Volume 85 (2000), pp. 5312-5315
[80] Data registration and selective single-molecule analysis using multi-parameter fluorescence detection, J. Biotechnol., Volume 86 (2001), pp. 163-180
[81] Polarization spectroscopy of single fluorescent molecules, J. Phys. Chem. B, Volume 103 (1999), pp. 6839-6850
[82] Protein structural dynamics by single-molecule fluorescence polarization, Prog. Biophys. Mol. Biol., Volume 74 (2000), pp. 1-35
[83] Single molecule polarization spectroscopy: pentacene in p-terphenyl, Chem. Phys., Volume 211 (1996), pp. 421-430
[84] Hindered rotational diffusion and rotational jumps of single molecules, Phys. Rev. Lett., Volume 80 (1998), pp. 2093-2096
[85] Simultaneous imaging of individual molecules aligned both parallel and perpendicular to the optic axis, Phys. Rev. Lett., Volume 81 (1998), pp. 5322-5325
[86] Imaging three-dimensional single molecule orientations, J. Phys. Chem. B, Volume 103 (1999), pp. 11237-11241
[87] Orientational imaging of single molecules by annular illumination, Phys. Rev. Lett., Volume 85 (2000), pp. 4482-4485
[88] Original tools for single-molecule spectroscopy, Single Molecules, Volume 2 (2001), pp. 241-249
[89] The renaissance of fluorescence resonance energy transfer, Nature Struct. Biol., Volume 7 (1999), pp. 730-734
[90] Single-molecule fluorescence resonance energy transfer, Methods, Volume 25 (2001), pp. 78-86
[91] Energy transfer: A spectroscopic ruler, Proc. Nat. Acad. Sci. USA, Volume 58 (1967), pp. 719-726
[92] Evaluation of the distribution of distances between energy donors and acceptors by fluorescence decay, Proc. Nat. Acad. Sci. USA, Volume 69 (1972), pp. 2273-2277
[93] Room-temperature fluorescence imaging and spectroscopy of single molecules by two-photon excitation, J. Phys. Chem. A, Volume 101 (1997), pp. 7019-7023
[94] Two-photon fluorescence excitation cross sections of biomolecular probes from 690 to 960 nm, Appl. Opt., Volume 37 (1998), pp. 7352-7356
[95] Photobleaching and stabilization of fluorophores used for single-molecule analysis with one- and two-photon excitation, Appl. Phys. B, Volume 73 (2001), pp. 829-837
[96] Probing single molecules and single nanoparticles with surface-enhanced raman scattering, Science, Volume 275 (1997), pp. 1102-1106
[97] Surface-enhanced non-linear Raman scattering at the single-molecule level, Chem. Phys., Volume 247 (1999), pp. 155-162
[98] Single-molecule kinetics of interfacial electron transfer, J. Phys. Chem. B, Volume 101 (1997), pp. 2753-2757
[99] Electron transfers in chemistry and biology, Biochem. Biophys. Acta, Volume 811 (1985), pp. 265-322
[100] Nanovid tracking: a new automatic method for the study of mobility in living cells based on colloidal gold and video microscopy, Biophys. J., Volume 52 (1987), pp. 775-782
[101] Tracking kinesin-driven movements with nanometer scale precision, Nature, Volume 331 (1988), pp. 450-453
[102] Position measurement with a noise-limited instrument, Rev. Sci. Instrum., Volume 57 (1986), pp. 1152-1157
[103] Determination of three-dimensional imaging properties of a light microscope system, Biophys. J., Volume 57 (1990), pp. 325-333
[104] Tracking of single fluorescent particles in three dimensions: use of cylindrical optics to encode particle position, Biophys. J., Volume 67 (1994), pp. 1291-1300
[105] Principles of Optics, Cambridge University Press, 1999
[106] Proposed method for molecular optical imaging, Opt. Lett., Volume 20 (1995), pp. 237-239
[107] Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission, Proc. Nat. Acad. Sci. USA, Volume 97 (2000), pp. 8206-8210
[108] Ultrahigh-resolution colocalization of spectrally resolvable point-like fluorescent probes, Methods, Volume 25 (2001), pp. 87-102
[109] Magnetic resonance of a single molecular spin, Nature, Volume 363 (1993), pp. 242-244
[110] Optical detection of magnetic resonance in a single molecule, Nature, Volume 363 (1993), pp. 244-245
[111] Tark effect on single molecules of dibenzanthanthrene in a naphthalene crystal and in a n-hexadecane Shpol'skii matrix, J. Phys. Chem. A, Volume 103 (1999), pp. 2429-2434
[112] Lassifying the photophysical dynamics of single- and multiple-chromophoric molecules by single molecule spectroscopy, J. Phys. Chem. A, Volume 102 (1998), pp. 7564-7575
[113] Single molecule spectroscopy on the light-harvesting complex II of higher plants, Biophys. J., Volume 81 (2001), pp. 556-562
[114] Fluorescence behavior of single-molecule pH-sensors, Single Molecules, Volume 1 (2000), pp. 17-23
[115] Folding dynamics of single GCN4 peptides by fluorescence resonant energy transfer confocal microscopy, Chem. Phys., Volume 247 (1999), pp. 69-83
[116] A single-molecule study of RNA catalysis and folding, Science, Volume 288 (2000), pp. 2048-2051
[117] Substeps within the 8-nm step of the ATPase cycle of single kinesin molecules, Nature Cell Biol., Volume 3 (2001), pp. 425-428
[118] ADP-induced rocking of the kinesin motor domain revealed by single-molecule fluorescence polarization microscopy, Nature Struct. Biol., Volume 8 (2001), pp. 540-544
[119] Stepping rotation of F1-ATPase visualized through angle-resolved single-fluorophore imaging, Proc. Nat. Acad. Sci. USA, Volume 97 (2000)
[120] Single-molecule observation of protein–protein interactions in the chaperonin system, Nature Biotech., Volume 19 (2001), pp. 861-865
[121] Resolution of distinct rotational substeps by submillisecond kinetic analysis of F1-ATPase, Nature, Volume 410 (2001), pp. 898-904
[122] Single-molecule protein folding: Diffusion fluorescence resonance energy transfer studies of the denaturation of chymotrypsin inhibitor 2, Proc. Nat. Acad. Sci. USA, Volume 97 (2000), pp. 5179-5184
[123] Single-molecule imaging of EGFR signalling on the surface of living cells, Nature Cell Biol., Volume 2 (2000), pp. 168-172
[124] Single molecule imaging of green fluorescent proteins in living cells: E-cadherin forms oligomers on the free cell surface, Biophys. J., Volume 80 (2001), pp. 2667-2677
[125] Single-molecule imaging of L-type Ca2+ channels in live cells, Biophys. J., Volume 81 (2001), pp. 2639-2646
[126] Single-molecule analysis of signaling in dictyostelium cells, Science, Volume 294 (2001), pp. 864-867
[127] Real-time single-molecule imaging of the infection pathway of an adeno-associated virus, Science, Volume 294 (2001), pp. 1929-1932
[128] Total internal reflection fluorescence microscopy in cell biology, Traffic, Volume 2 (2001), pp. 764-774
Cité par Sources :
Commentaires - Politique