Subdiffraction-resolution fluorescence imaging of proteins in the mitochondrial inner membrane with photoswitchable fluorophores

van de Linde S, Sauer M, Heilemann M (2008)

Zeitschriftenaufsatz | Veröffentlicht | Englisch
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van de Linde, Sebastian; Sauer, MarkusUniBi; Heilemann, MikeUniBi
Abstract / Bemerkung
Understanding the structural organization of biomolecules in cells, sub-cellular compartments or membranes requires non-invasive methods of observation that provide high spatial resolution. Recent advancements in fluorescence microscopy paved the way for novel super-resolution observations with an optical resolution well below the diffraction barrier of light. Here, we demonstrate that commercially available standard fluorescent probes, i.e. Alexa 647 labeled antibodies, can be used as efficient photoswitches. In combination with localization microscopy approaches the method is ideally suited to study the spatial organization of proteins in sub-cellular structures and membranes. The simplicity of the method lies in the fact that standard immunocytochemistry assays together with photoswitchable carbocyanine fluorophores and conventional total internal reflection fluorescence (TIRF) microscopy can be used to achieve a lateral resolution of similar to 20 nm. We demonstrate subdiffraction-resolution fluorescence imaging of intracellular F0F1-ATP synthase and cytochrome c oxidase in the inner membrane of mitochondria. Besides the high localization precision of individual proteins we demonstrate how quantitative data, i.e. the protein distribution in the membrane, can be derived and compared. (C) 2008 Elsevier Inc. All rights reserved.
Subdiffraction-resolution; dSTORM; Super-resolution; Single-molecule spectroscopy; Mitochondria; Molecular quantification
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van de Linde S, Sauer M, Heilemann M. Subdiffraction-resolution fluorescence imaging of proteins in the mitochondrial inner membrane with photoswitchable fluorophores. JOURNAL OF STRUCTURAL BIOLOGY. 2008;164(3):250-254.
van de Linde, S., Sauer, M., & Heilemann, M. (2008). Subdiffraction-resolution fluorescence imaging of proteins in the mitochondrial inner membrane with photoswitchable fluorophores. JOURNAL OF STRUCTURAL BIOLOGY, 164(3), 250-254.
van de Linde, S., Sauer, M., and Heilemann, M. (2008). Subdiffraction-resolution fluorescence imaging of proteins in the mitochondrial inner membrane with photoswitchable fluorophores. JOURNAL OF STRUCTURAL BIOLOGY 164, 250-254.
van de Linde, S., Sauer, M., & Heilemann, M., 2008. Subdiffraction-resolution fluorescence imaging of proteins in the mitochondrial inner membrane with photoswitchable fluorophores. JOURNAL OF STRUCTURAL BIOLOGY, 164(3), p 250-254.
S. van de Linde, M. Sauer, and M. Heilemann, “Subdiffraction-resolution fluorescence imaging of proteins in the mitochondrial inner membrane with photoswitchable fluorophores”, JOURNAL OF STRUCTURAL BIOLOGY, vol. 164, 2008, pp. 250-254.
van de Linde, S., Sauer, M., Heilemann, M.: Subdiffraction-resolution fluorescence imaging of proteins in the mitochondrial inner membrane with photoswitchable fluorophores. JOURNAL OF STRUCTURAL BIOLOGY. 164, 250-254 (2008).
van de Linde, Sebastian, Sauer, Markus, and Heilemann, Mike. “Subdiffraction-resolution fluorescence imaging of proteins in the mitochondrial inner membrane with photoswitchable fluorophores”. JOURNAL OF STRUCTURAL BIOLOGY 164.3 (2008): 250-254.

45 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Localization-based super-resolution imaging meets high-content screening.
Beghin A, Kechkar A, Butler C, Levet F, Cabillic M, Rossier O, Giannone G, Galland R, Choquet D, Sibarita JB., Nat Methods 14(12), 2017
PMID: 29083400
Imaging cellular structures in super-resolution with SIM, STED and Localisation Microscopy: A practical comparison.
Wegel E, Göhler A, Lagerholm BC, Wainman A, Uphoff S, Kaufmann R, Dobbie IM., Sci Rep 6(), 2016
PMID: 27264341
Methodology for Quantitative Characterization of Fluorophore Photoswitching to Predict Superresolution Microscopy Image Quality.
Bittel AM, Nickerson A, Saldivar IS, Dolman NJ, Nan X, Gibbs SL., Sci Rep 6(), 2016
PMID: 27412307
Endosome-mitochondria interactions are modulated by iron release from transferrin.
Das A, Nag S, Mason AB, Barroso MM., J Cell Biol 214(7), 2016
PMID: 27646275
Superresolution microscopy reveals spatial separation of UCP4 and F0F1-ATP synthase in neuronal mitochondria.
Klotzsch E, Smorodchenko A, Löfler L, Moldzio R, Parkinson E, Schütz GJ, Pohl EE., Proc Natl Acad Sci U S A 112(1), 2015
PMID: 25535394
Super-resolution microscopy of mitochondria.
Jakobs S, Wurm CA., Curr Opin Chem Biol 20(), 2014
PMID: 24769752
Imaging fenestrations in liver sinusoidal endothelial cells by optical localization microscopy.
Mönkemöller V, Schüttpelz M, McCourt P, Sørensen K, Smedsrød B, Huser T., Phys Chem Chem Phys 16(24), 2014
PMID: 24830784
Single-molecule coordinate-based analysis of the morphology of HIV-1 assembly sites with near-molecular spatial resolution.
Malkusch S, Muranyi W, Müller B, Kräusslich HG, Heilemann M., Histochem Cell Biol 139(1), 2013
PMID: 22910843
Superresolution microscopy in heart - cardiac nanoscopy.
Kohl T, Westphal V, Hell SW, Lehnart SE., J Mol Cell Cardiol 58(), 2013
PMID: 23219451
Fluorescence nanoscopy. Methods and applications.
Requejo-Isidro J., J Chem Biol 6(3), 2013
PMID: 24432127
Photoactivatable synthetic fluorophores.
Raymo FM., Phys Chem Chem Phys 15(36), 2013
PMID: 23780303
Test samples for optimizing STORM super-resolution microscopy.
Metcalf DJ, Edwards R, Kumarswami N, Knight AE., J Vis Exp (79), 2013
PMID: 24056752
Quantitative morphological analysis of arrestin2 clustering upon G protein-coupled receptor stimulation by super-resolution microscopy.
Truan Z, Tarancón Díez L, Bönsch C, Malkusch S, Endesfelder U, Munteanu M, Hartley O, Heilemann M, Fürstenberg A., J Struct Biol 184(2), 2013
PMID: 24091038
Live-cell dSTORM of cellular DNA based on direct DNA labeling.
Benke A, Manley S., Chembiochem 13(2), 2012
PMID: 22213360
Multi-colour direct STORM with red emitting carbocyanines.
Lampe A, Haucke V, Sigrist SJ, Heilemann M, Schmoranzer J., Biol Cell 104(4), 2012
PMID: 22187967
Fluorophore targeting to cellular proteins via enzyme-mediated azide ligation and strain-promoted cycloaddition.
Yao JZ, Uttamapinant C, Poloukhtine A, Baskin JM, Codelli JA, Sletten EM, Bertozzi CR, Popik VV, Ting AY., J Am Chem Soc 134(8), 2012
PMID: 22239252
Super-resolution microscopy: a comparative treatment.
Kasuboski JM, Sigal YJ, Joens MS, Lillemeier BF, Fitzpatrick JA., Curr Protoc Cytom Chapter 2(), 2012
PMID: 23042525
Molecular strategies to read and write at the nanoscale with far-field optics.
Cusido J, Impellizzeri S, Raymo FM., Nanoscale 3(1), 2011
PMID: 20936237
Direct stochastic optical reconstruction microscopy with standard fluorescent probes.
van de Linde S, Löschberger A, Klein T, Heidbreder M, Wolter S, Heilemann M, Sauer M., Nat Protoc 6(7), 2011
PMID: 21720313
Real-time computation of subdiffraction-resolution fluorescence images.
Wolter S, Schüttpelz M, Tscherepanow M, VAN DE Linde S, Heilemann M, Sauer M., J Microsc 237(1), 2010
PMID: 20055915
A new wave of cellular imaging.
Toomre D, Bewersdorf J., Annu Rev Cell Dev Biol 26(), 2010
PMID: 20929313
Spiropyrans as molecular optical switches.
Seefeldt B, Kasper R, Beining M, Mattay J, Arden-Jacob J, Kemnitzer N, Drexhage KH, Heilemann M, Sauer M., Photochem Photobiol Sci 9(2), 2010
PMID: 20126797
Higher resolution in localization microscopy by slower switching of a photochromic protein.
Mizuno H, Dedecker P, Ando R, Fukano T, Hofkens J, Miyawaki A., Photochem Photobiol Sci 9(2), 2010
PMID: 20126801
Subdiffraction-resolution fluorescence microscopy of myosin-actin motility.
Endesfelder U, van de Linde S, Wolter S, Sauer M, Heilemann M., Chemphyschem 11(4), 2010
PMID: 20186905
Subdiffraction fluorescence imaging of biomolecular structure and distributions with quantum dots.
Heidbreder M, Endesfelder U, van de Linde S, Hennig S, Widera D, Kaltschmidt B, Kaltschmidt C, Heilemann M., Biochim Biophys Acta 1803(10), 2010
PMID: 20600360
Make them blink: probes for super-resolution microscopy.
Vogelsang J, Steinhauer C, Forthmann C, Stein IH, Person-Skegro B, Cordes T, Tinnefeld P., Chemphyschem 11(12), 2010
PMID: 20632356
Dynamic superresolution imaging of endogenous proteins on living cells at ultra-high density.
Giannone G, Hosy E, Levet F, Constals A, Schulze K, Sobolevsky AI, Rosconi MP, Gouaux E, Tampé R, Choquet D, Cognet L., Biophys J 99(4), 2010
PMID: 20713016
Surfing on a new wave of single-molecule fluorescence methods.
Hohlbein J, Gryte K, Heilemann M, Kapanidis AN., Phys Biol 7(3), 2010
PMID: 20686191
The rylene colorant family--tailored nanoemitters for photonics research and applications.
Weil T, Vosch T, Hofkens J, Peneva K, Müllen K., Angew Chem Int Ed Engl 49(48), 2010
PMID: 20973116
Advances in light microscopy for neuroscience.
Wilt BA, Burns LD, Wei Ho ET, Ghosh KK, Mukamel EA, Schnitzer MJ., Annu Rev Neurosci 32(), 2009
PMID: 19555292
Super-resolution imaging with small organic fluorophores.
Heilemann M, van de Linde S, Mukherjee A, Sauer M., Angew Chem Int Ed Engl 48(37), 2009
PMID: 19670280
Multicolor photoswitching microscopy for subdiffraction-resolution fluorescence imaging.
van de Linde S, Endesfelder U, Mukherjee A, Schüttpelz M, Wiebusch G, Wolter S, Heilemann M, Sauer M., Photochem Photobiol Sci 8(4), 2009
PMID: 19337659
Dual color localization microscopy of cellular nanostructures.
Gunkel M, Erdel F, Rippe K, Lemmer P, Kaufmann R, Hörmann C, Amberger R, Cremer C., Biotechnol J 4(6), 2009
PMID: 19548231
Biomimetic approaches for studying membrane processes.
Jelinek R, Silbert L., Mol Biosyst 5(8), 2009
PMID: 19603114
Super-resolution imaging of DNA labelled with intercalating dyes.
Flors C, Ravarani CN, Dryden DT., Chemphyschem 10(13), 2009
PMID: 19554598

22 References

Daten bereitgestellt von Europe PubMed Central.

Image processing with ImageJ
Abramoff, Biophotonics Int. 11(), 2004
Short-range spectroscopic ruler based on a single-molecule optical switch.
Bates M, Blosser TR, Zhuang X., Phys. Rev. Lett. 94(10), 2005
PMID: 15783528
Multicolor super-resolution imaging with photo-switchable fluorescent probes.
Bates M, Huang B, Dempsey GT, Zhuang X., Science 317(5845), 2007
PMID: 17702910
Imaging intracellular fluorescent proteins at nanometer resolution.
Betzig E, Patterson GH, Sougrat R, Lindwasser OW, Olenych S, Bonifacino JS, Davidson MW, Lippincott-Schwartz J, Hess HF., Science 313(5793), 2006
PMID: 16902090
Supramolecular structure of the mitochondrial oxidative phosphorylation system.
Boekema EJ, Braun HP., J. Biol. Chem. 282(1), 2006
PMID: 17102127
The ATP synthase--a splendid molecular machine.
Boyer PD., Annu. Rev. Biochem. 66(), 1997
PMID: 9242922
Rows of ATP synthase dimers in native mitochondrial inner membranes.
Buzhynskyy N, Sens P, Prima V, Sturgis JN, Scheuring S., Biophys. J. 93(8), 2007
PMID: 17557793
Trans lipids: the free radical path.
Chatgilialoglu C, Ferreri C., Acc. Chem. Res. 38(6), 2005
PMID: 15966710
Two-color far-field fluorescence nanoscopy.
Donnert G, Keller J, Wurm CA, Rizzoli SO, Westphal V, Schonle A, Jahn R, Jakobs S, Eggeling C, Hell SW., Biophys. J. 92(8), 2007
PMID: 17307826
A stroboscopic approach for fast photoactivation-localization microscopy with Dronpa mutants.
Flors C, Hotta J, Uji-i H, Dedecker P, Ando R, Mizuno H, Miyawaki A, Hofkens J., J. Am. Chem. Soc. 129(45), 2007
PMID: 17956094
The cristal membrane of mitochondria is the principal site of oxidative phosphorylation.
Gilkerson RW, Selker JM, Capaldi RA., FEBS Lett. 546(2-3), 2003
PMID: 12832068
Carbocyanine dyes as efficient reversible single-molecule optical switch.
Heilemann M, Margeat E, Kasper R, Sauer M, Tinnefeld P., J. Am. Chem. Soc. 127(11), 2005
PMID: 15771514
Subdiffraction-resolution fluorescence imaging with conventional fluorescent probes.
Heilemann M, van de Linde S, Schuttpelz M, Kasper R, Seefeldt B, Mukherjee A, Tinnefeld P, Sauer M., Angew. Chem. Int. Ed. Engl. 47(33), 2008
PMID: 18646237
Far-field optical nanoscopy.
Hell SW., Science 316(5828), 2007
PMID: 17525330
Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy.
Huang B, Wang W, Bates M, Zhuang X., Science 319(5864), 2008
PMID: 18174397
4Pi microscopy of quantum dot-labeled cellular structures.
Medda R, Jakobs S, Hell SW, Bewersdorf J., J. Struct. Biol. 156(3), 2006
PMID: 17045487
Three-dimensional structure of the respiratory chain supercomplex I1III2IV1 from bovine heart mitochondria.
Schafer E, Dencher NA, Vonck J, Parcej DN., Biochemistry 46(44), 2007
PMID: 17927210
Architecture of active mammalian respiratory chain supercomplexes.
Schafer E, Seelert H, Reifschneider NH, Krause F, Dencher NA, Vonck J., J. Biol. Chem. 281(22), 2006
PMID: 16551638
Respiratory chain supercomplexes of mitochondria and bacteria.
Schagger H., Biochim. Biophys. Acta 1555(1-3), 2002
PMID: 12206908
Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy.
Schermelleh L, Carlton PM, Haase S, Shao L, Winoto L, Kner P, Burke B, Cardoso MC, Agard DA, Gustafsson MG, Leonhardt H, Sedat JW., Science 320(5881), 2008
PMID: 18535242
Dimer ribbons of ATP synthase shape the inner mitochondrial membrane.
Strauss M, Hofhaus G, Schroder RR, Kuhlbrandt W., EMBO J. 27(7), 2008
PMID: 18323778
Precise nanometer localization analysis for individual fluorescent probes.
Thompson RE, Larson DR, Webb WW., Biophys. J. 82(5), 2002
PMID: 11964263


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