Chemical Society Reviews, Год журнала: 2022, Номер 51(24), С. 9882 - 9916
Опубликована: Янв. 1, 2022
Following an overview of the approaches and techniques used to acheive super-resolution microscopy, this review presents advantages supplied by nanoparticle based probes for these applications.
Язык: Английский
Journal of Biological Chemistry, Год журнала: 2021, Номер 297(1), С. 100791 - 100791
Опубликована: Май 18, 2021
Super-resolution microscopy has become an increasingly popular and robust tool across the life sciences to study minute cellular structures processes. However, with increasing number of available super-resolution techniques come increased complexity burden choice in planning imaging experiments. Choosing right technique answer a given biological question is vital for understanding interpreting relevance. This often-neglected complex task that should take into account well-defined criteria (e.g., sample type, structure size, requirements). Trade-offs different capabilities are inevitable; thus, many researchers still find it challenging select most suitable will best their question. review aims provide overview clarify concepts underlying commonly as well guide through all aspects be considered before opting technique. Optical long been indispensable research. Beginning development cell theory Hooke's iconic Micrographia 1665 (1Hooke R. Micrographia, or Some Physiological Descriptions Minute Bodies Made by Magnifying Glasses, Observations Inquiries Thereupon. Martyn, JAllestry, J, London1665Crossref Google Scholar), have underpinned field's exciting discoveries, from evolution (2Jardine B. Between Beagle barnacle: Darwin's microscopy, 1837-1854.Stud. Hist. Philos. Sci. 2009; 40: 382-395Crossref PubMed Scopus (6) Scholar) observing molecular machines (3Yasuda Noji H. Kinosita K. Yoshida M. F1-ATPase highly efficient motor rotates discrete 120 degree steps.Cell. 1998; 93: 1117-1124Abstract Full Text PDF (690) Scholar). Despite successes, plagued limitations imposed laws physics, preventing these optical systems resolving features below certain known diffraction limit. The limit stems combination wave nature light combined inability our focus waves single point diameter. limit, which determines minimum distance needs separate two points order them resolved system, was first defined 1873 German physicist, Ernst Abbe (4Abbe E. Beiträge zur Theorie des Mikroskops und der mikroskopischen Wahrnehmung.Archiv f. Mikrosk. Anatomie. 1873; 9: 413-418Crossref Scholar, 5Abbe A contribution microscope microscopic vision.in: Proceedings Bristol Naturalists' Society. vol. 1. Williams & Northgate, London, UK1876: 200-261Google dependent on wavelength light, refractive index medium travels, angles diffracted can collected objective. latter factors used calculate numerical aperture (NA) specific objectives, represents unitless measure range objective accept incoming thus affects resolution achievable Indeed, showed roughly equal (λ) divided twice NA lateral resolution, 2λ/NA2 axial resolution. means standard fluorescence such confocal operating visible spectrum, limited around 170 250 nm laterally 470 670 axially, when detecting wavelengths between 700 nm. Prior 2000, only able image subdiffraction were near-field-based techniques, use nanometric detectors placed very close detect evanescent (6Hecht Sick Wild U.P. Deckert V. Zenobi Martin O.J.F. Pohl D.W. Scanning near-field probes: Fundamentals applications.J. Chem. Phys. 2000; 112: 7761Crossref Since turn millennium, however, far-field emerged overcome this review. broadly split categories: super-resolved ensemble improve overall structures, fluorophore localizations individual fluorescent molecules build up structure. REversible Saturable OpticaL Fluorescence Transition (RESOLFT) include stimulated emission depletion (STED) similar ground state depletion/RESOLFT structured illumination (SIM), pixel reassignment arguably preparation–based expansion (ExM). Single-fluorophore meanwhile, collective single-molecule localization (SMLM) notably direct stochastic reconstruction (dSTORM), photoactivated (PALM), DNA-based accumulation nanoscale topography (DNA-PAINT). inception, steadily gained popularity, leading ever discoveries previously eluded detection. impact research 2014 Nobel Prize Chemistry awarded Stefan W. Hell, Eric Betzig, William Moerner; Hell STED Betzig Moerner work PALM subsequent (7Ehrenberg Scientific Background 2014. Royal Swedish Academy Sciences, Stockholm, Sweden2014Google Given wealth nontrivial information address therefore insight important considerations selecting problem at hand. does not in-depth explanation each but rather seeks help biologists assess answering compatible needs. laser scanning–based technique, achieved addition high-powered torus-shaped lasers. lasers aligned excitation beam deplete overlapping region (Fig. 1Ai) (8Hell S.W. Wichmann J. Breaking emission: Stimulated-emission-depletion microscopy.Opt. Lett. 1994; 19: 780-782Crossref 9Klar T.A. Engel Abbe's beams various shapes.Phys. Rev. E Stat. Nonlin. Soft Matter 2001; 64: 066613Crossref interruption internal conversion process undergone excited electrons, high-energy forcing immediately return state. results release photon used, easily removed filters 1Aii) initially implemented using two-photon now commercially continuous (CW) pulsed longer pulse lengths CW simplify implementation removing need synchronization pulses (10Willig K.I. Harke Medda beams.Nat. Methods. 2007; 4: 915-918Crossref (393) Scholar); being exposed pulses, contribute formation, photobleaching. Unlike STED, where depleting photons arrive shortly after sample, implementations suffer added issue lower instantaneous intensities meaning greater enough depleted, degrading attainable (11Vicidomini G. Schönle A. Ta Han K.Y. Moneron Eggeling C. nanoscopy time-gated detection: Theoretical experimental aspects.PLoS One. 2013; 8e54421Crossref (101) rise short-lifetime emitted time gating they unlikely had sufficient opportunity undergo 12Moffitt J.R. Osseforth Michaelis Time-gating improves spatial Express. 2011; 4242-4254Crossref (0) Pulsed also benefit gating. Commercial ~1 ns length, compared picosecond femtosecond 2-photon used. length allows power spread over duration, reducing photobleaching (13Dyba Photostability marker under excited-state emission.Appl. Opt. 2003; 42: 5123-5129Crossref issues observed shorter including polarization effects (14Westphal Nanoscale focal plane microscope.Phys. 2005; 94: 143903Crossref (378) jitter (15Westphal Blanca C.M. Dyba Kastrup L. Laser-diode-stimulated microscopy.Appl. 82: 3125Crossref efficiency slightly lengths, reduction eliminated (16Göttfert F. Wurm C.A. Mueller Berning S. Cordes V.C. Honigmann Coaligned dual-channel diffusion analysis 20 resolution.Biophys. 105: L01-L03Abstract (190) Details within samples detected microscopes combinations frequencies. optics inherently frequencies highest-frequency (i.e., highest-resolution details sample) lost. SIM advantage phenomenon Moiré effect, whereby interference differing high-frequency patterns overlaid produce sum (very high) difference (low). Therefore, pattern just microscope, possible move sampling microscope. By controlling orientation phase pattern, original back-calculated 1B) (17Gustafsson M.G. Surpassing factor microscopy.J. Microsc. 198: 82-87Crossref (2347) To achieve full reconstruction, several images positions orientations reconstruct final Laser scanning confocal-based generally acquire data onto detector comprising pixel. Pixel combining point-scanning multipixel array create higher frequency component image. central adapted plays role pinhole, additional pixels allow more structural acquisition. Perhaps Zeiss's Airyscan, uses composed 32 elements, equivalent 0.2 airy unit pinhole 1C) (18Huff Airyscan ZEISS: Confocal improved signal-to-noise ratio super-resolution.Nat. 2015; 12: i-iiCrossref Each element acts captured times than once. correcting displacement axis, reconstructed ExM relies adapting surpass physical itself, increases effective diffraction-limited embedding hyperswellable hydrogel chains cross-linking agents, creating extremely dense linked mesh (19Chen Tillberg P.W. Boyden E.S. imaging. Expansion microscopy.Science. 347: 543-548Crossref (568) During gelation, proteins fluorophores anchored gel. permeated then mechanically homogenized (high temperature, detergents, proteases, etc) avoid resistance expansion, water triggers swelling polymer, 100× its size 1D) Expanded imaged widefield SMLM numerous varied, rely same principle differentiate space; distances, cannot differentiated fluorescing 2Ai) pinpointed if fluoresce one 2Aii). Individually emit hundreds thousands photons, producing intensity multiple another, overlap (20Thompson R.E. Larson D.R. Webb W.W. Precise nanometer probes.Biophys. 2002; 2775-2783Abstract 21Endesfelder U. Malkusch Fricke Heilemann simple method estimate average precision experiment.Histochem. Cell Biol. 2014; 141: 629-638Crossref In absence neighboring fluorophores, limited, distribution emits calculation subpixel All having dyes switch off states, achieves unique way. dSTORM PALM. specialized buffers drive organic long-lived dark without returning 2B) (22Heilemann Margeat Kasper Sauer Tinnefeld P. Carbocyanine reversible switch.J. Am. Soc. 127: 3801-3806Crossref (315) 23Heilemann van de Linde Schüttpelz Seefeldt Mukherjee Subdiffraction-resolution conventional probes.Angew. Int. Ed. Engl. 2008; 47: 6172-6176Crossref (1236) optimal buffer composition selected maximize rate entry minimize rates escape inclusion dissolved oxygen. If sufficiently large proportion reside any time, momentarily emissions isolated. publications focused further subject (24Vogelsang Steinhauer Person oxidizing system minimizes blinking dyes.Angew. 5465-5469Crossref 25Nahidiazar Agronskaia A.V. Broertjes den Broek Jalink Optimizing conditions demanding multi-color super microscopy.PLoS 2016; 11e0158884Crossref (82) 26Goossen-Schmidt N.C. Schnieder Hüve Klingauf Switching behaviour glycerol-containing buffer.Sci. Rep. 2020; 10: 13746Crossref (1) PALM, other hand, photoactivatable photoswitchable genetically encoded. common natively found state, (27Betzig Patterson G.H. Sougrat Lindwasser O.W. Olenych Bonifacino J.S. Davidson M.W. Lippincott-Schwartz Hess H.F. Imaging intracellular resolution.Science. 2006; 313: 1642-1645Crossref (5805) Illumination activation wavelength, often UV, causes conformational change 2C) Photoswitchable comparison, normally photoconvert species form, way (28Li Vaughan J.C. Switchable microscopy.Chem. 2018; 118: 9412-9454Crossref (99) Detection cycling low levels subset allowing bleach Although typically brighter photostable proteins, there extreme variability dyes, properties depends upon (29Dempsey G.T. Chen K.H. Bates Zhuang X. Evaluation performance localization-based imaging.Nat. 8: 1027-1036Crossref (828) Choice dye directly affect accuracy Fluctuation-based Fluctuation (SOFI) (30Dertinger T. Colyer Iyer Weiss Enderlein Fast, background-free, 3D fluctuation (SOFI).Proc. Natl. Acad. 106: 22287-22292Crossref (698) Super-Resolution Radial Fluctuations (31Gustafsson N. Culley Ashdown Owen D.M. Pereira P.M. Henriques Fast live-cell ImageJ radial fluctuations.Nat. Commun. 7: 12471Crossref (243) natural variations transition nonfluorescent states 2D). fluctuation-based localize instead calculates statistical entire frame 31Gustafsson probes fluctuate, better be, samples; prepared Minimal flux (MINFLUX) combines For MINFLUX imaging, required blink center (32Balzarotti Eilers Y. Gwosch K.C. Gynnå A.H. Westphal Stefani F.D. Elf Nanometer tracking minimal fluxes.Science. 2017; 355: 606-612Crossref (404) exactly centre excited, pinpoint precise coordinates Excitation decrease position approaches 2E) contrast approach efficient, requiring 100-fold fewer resolutions state-of-the-art (33Masullo L.A. Steiner Zähringer Lopez L.F. Bohlen Richter Cole interleaved MINFLUX.Nano 2021; 21: 840-846Crossref (10) Moreover, far surpasses both (34Gwosch Pape J.K. Balzarotti Hoess Ellenberg Ries delivers multicolor cells.Nat. 17: 217-224Crossref (156) relatively computationally intensive and, produced December 2019, widely available. encompass distinct strengths limitations. When designing experiment, appropriately core Biological questions investigated categorized those concerned defining high-resolution interest, dynamics, interactions. following sections discuss categories each. obvious frequent application addressing subcellular colocalization, require beyond While labeled target localized improvements always met trade-off abilities temporal avoidance phototoxicity/photobleaching. It essential consider requirements experiment determine data. detailed discussed later, simplified flow diagram appropriate presented Figure 3. aspect taken consideration smallest distinguished entities. varies 4)—from 1.4× improvement resolu
Язык: Английский
Процитировано
125
Nature Methods, Год журнала: 2022, Номер 19(9), С. 1072 - 1075
Опубликована: Сен. 1, 2022
Abstract MINimal fluorescence photon FLUXes (MINFLUX) nanoscopy, providing photon-efficient fluorophore localizations, has brought about three-dimensional resolution at nanometer scales. However, by using an intrinsic on–off switching process for single separation, initial MINFLUX implementations have been limited to two color channels. Here we show that can be effectively combined with sequentially multiplexed DNA-based labeling (DNA-PAINT), expanding nanoscopy multiple molecular targets. Our method is exemplified three-color recordings of mitochondria in human cells.
Язык: Английский
Процитировано
79
Nature Methods, Год журнала: 2022, Номер 19(8), С. 986 - 994
Опубликована: Авг. 1, 2022
Advances in super-resolution microscopy have demonstrated single-molecule localization precisions of a few nanometers. However, translation such high into sub-10-nm spatial resolution biological samples remains challenging. Here we show that resonance energy transfer between fluorophores separated by less than 10 nm results accelerated fluorescence blinking and consequently lower probabilities impeding imaging. We demonstrate time-resolved detection combination with photoswitching fingerprint analysis can be used to determine the number distance even spatially unresolvable range. In genetic code expansion unnatural amino acids bioorthogonal click labeling small fluorophores, advantageously reveal information about present their distances range cells.
Язык: Английский
Процитировано
56
eLight, Год журнала: 2022, Номер 2(1)
Опубликована: Март 25, 2022
Abstract Achieving 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 (2p-MINFLUX) could double PSF engineering nonlinear effect. Cramér-Rao Bound (CRB) studied as maximum precision, and CRB halved compared single-photon (1p-MINFLUX) in all three dimensions. Meanwhile, order achieve same 1p-MINFLUX, 2p-MINFLUX requires only 1/4 photons. Exploiting simultaneous excitation multiple fluorophore species, may have potential registration-free nanoscopy multicolor tracking.
Язык: Английский
Процитировано
43
Light Science & Applications, Год журнала: 2022, Номер 11(1)
Опубликована: Июнь 30, 2022
Localization of single fluorescent emitters is key for physicochemical and biophysical measurements at the nanoscale beyond ensemble averaging. Examples include single-molecule tracking super-resolution imaging by localization microscopy. Among numerous methods available, MINFLUX outstands achieving a ~10-fold improvement in resolution over wide-field camera-based approaches, reaching molecular scale moderate photon counts. Widespread application related has been hindered technical complexity setups. Here, we present RASTMIN, method based on raster scanning light pattern comprising minimum intensity. RASTMIN delivers ~1-2 nm precision with usual fluorophores easily implementable standard confocal microscope few modifications. We demonstrate performance molecules DNA origami structures.
Язык: Английский
Процитировано
42
Nature Photonics, Год журнала: 2024, Номер 18(5), С. 478 - 484
Опубликована: Фев. 9, 2024
Язык: Английский
Процитировано
12
Annual Review of Physical Chemistry, Год журнала: 2024, Номер 75(1), С. 163 - 183
Опубликована: Фев. 16, 2024
By superlocalizing the positions of millions single molecules over many camera frames, a class super-resolution fluorescence microscopy methods known as single-molecule localization (SMLM) has revolutionized how we understand subcellular structures past decade. In this review, highlight emerging studies that transcend outstanding structural (shape) information offered by SMLM to extract and map physicochemical parameters in living mammalian cells at levels. encoding/decoding high-dimensional information-such emission excitation spectra, motion, polarization, lifetime, beyond-for every molecule, mass accumulating these measurements for molecules, such multidimensional multifunctional approaches open new windows into intracellular architectures dynamics, well their underlying biophysical rules, far beyond diffraction limit.
Язык: Английский
Процитировано
12
Advanced Materials, Год журнала: 2021, Номер 33(24)
Опубликована: Май 3, 2021
Abstract Graphene is considered a game‐changing material, especially for its mechanical and electrical properties. This work exploits that graphene almost transparent but quenches fluorescence in range up to ≈40 nm. as broadband unbleachable energy‐transfer acceptor without labeling, used precisely determine the height of molecules with respect graphene, visualize dynamics DNA nanostructures, orientation Förster‐type resonance energy transfer (FRET) pairs. Using origami nanopositioners, biosensing, single‐molecule tracking, PAINT super‐resolution <3 nm z ‐resolution are demonstrated. The examples shows potential graphene‐on‐glass coverslips versatile platform biophysics, microscopy.
Язык: Английский
Процитировано
53
Journal of the American Chemical Society, Год журнала: 2021, Номер 143(44), С. 18388 - 18393
Опубликована: Окт. 29, 2021
We propose a series of fluorescent dyes with hydrophilic carbamate caging groups that undergo rapid photoactivation under UV (≤400 nm) irradiation but do not spurious two-photon activation high-intensity (visible or infrared) light about twice the wavelength. The caged and labels derived therefrom display high water solubility convert upon into validated super-resolution live-cell-compatible fluorophores. In combination popular markers, multiple (up to six)-color images can be obtained stimulated emission depletion nanoscopy. Moreover, individual fluorophores localized precision <3 nm (standard deviation) using MINSTED MINFLUX techniques.
Язык: Английский
Процитировано
51
Journal of Cell Science, Год журнала: 2021, Номер 134(9), С. 1 - 17
Опубликована: Май 1, 2021
ABSTRACT A major focus of current biological studies is to fill the knowledge gaps between cell, tissue and organism scales. To this end, a wide array contemporary optical analytical tools enable multiparameter quantitative imaging live fixed cells, three-dimensional (3D) systems, tissues, organs organisms in context their complex spatiotemporal molecular features. In particular, modalities luminescence lifetime imaging, comprising fluorescence (FLI) phosphorescence microscopy (PLIM), synergy with Förster resonance energy transfer (FRET) assays, provide wealth information. On application side, endogenous molecules inside cells overexpressed fluorescent protein fusion biosensor constructs or probes delivered externally insights at multiple scales into protein–protein interaction networks, cellular metabolism, dynamics oxygen hypoxia, physiologically important ions, other physical physiological parameters. Luminescence offers unique window structural environment enabling new level functional analysis addition providing 3D spatially resolved longitudinal measurements that can range from microscopic macroscopic scale. We an overview summarize key applications tissues organisms.
Язык: Английский
Процитировано
44