Superresolution Imaging Using Single-molecule Localization

Overview

Journal Annu Rev Phys Chem

Publisher Annual Reviews

Specialty Chemistry

Date 2010 Jan 9

PMID 20055680

Citations 180

Authors

George Patterson

Michael Davidson

Suliana Manley

Jennifer Lippincott-Schwartz

Affiliations

Soon will be listed here.

Abstract

Superresolution imaging is a rapidly emerging new field of microscopy that dramatically improves the spatial resolution of light microscopy by over an order of magnitude (approximately 10-20-nm resolution), allowing biological processes to be described at the molecular scale. Here, we discuss a form of superresolution microscopy based on the controlled activation and sampling of sparse subsets of photoconvertible fluorescent molecules. In this single-molecule-based imaging approach, a wide variety of probes have proved valuable, ranging from genetically encodable photoactivatable fluorescent proteins to photoswitchable cyanine dyes. These have been used in diverse applications of superresolution imaging: from three-dimensional, multicolor molecule localization to tracking of nanometric structures and molecules in living cells. Single-molecule-based superresolution imaging thus offers exciting possibilities for obtaining molecular-scale information on biological events occurring at variable timescales.

Citing Articles

Electrochemically modulated single-molecule localization microscopy for imaging cytoskeletal protein structures.

Lei C, Hu D Nanophotonics. 2025; 14(4):459-470.

PMID: 39975638 PMC: 11834056. DOI: 10.1515/nanoph-2024-0559.

Super-resolution microscopy to study membrane nanodomains and transport mechanisms in the plasma membrane.

Zulueta Diaz Y, Arnspang E Front Mol Biosci. 2024; 11:1455153.

PMID: 39290992 PMC: 11405310. DOI: 10.3389/fmolb.2024.1455153.

Versatile nanobody-based approach to image, track and reconstitute functional Neurexin-1 in vivo.

Vicidomini R, Choudhury S, Han T, Nguyen T, Nguyen P, Opazo F Nat Commun. 2024; 15(1):6068.

PMID: 39025931 PMC: 11258300. DOI: 10.1038/s41467-024-50462-2.

Biospecific Chemistry for Covalent Linking of Biomacromolecules.

Cao L, Wang L Chem Rev. 2024; 124(13):8516-8549.

PMID: 38913432 PMC: 11240265. DOI: 10.1021/acs.chemrev.4c00066.

Advancements and Practical Considerations for Biophysical Research: Navigating the Challenges and Future of Super-resolution Microscopy.

Chen H, Yan G, Wen M, Brooks K, Zhang Y, Huang P Chem Biomed Imaging. 2024; 2(5):331-344.

PMID: 38817319 PMC: 11134610. DOI: 10.1021/cbmi.4c00019.

References

Gurskaya N, Verkhusha V, Shcheglov A, Staroverov D, Chepurnykh T, Fradkov A . Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light. Nat Biotechnol. 2006; 24(4):461-5. DOI: 10.1038/nbt1191. View

Flors C, Hotta J, Uji-I H, Dedecker P, Ando R, Mizuno H . A stroboscopic approach for fast photoactivation-localization microscopy with Dronpa mutants. J Am Chem Soc. 2007; 129(45):13970-7. DOI: 10.1021/ja074704l. View

Manley S, Gillette J, Patterson G, Shroff H, Hess H, Betzig E . High-density mapping of single-molecule trajectories with photoactivated localization microscopy. Nat Methods. 2008; 5(2):155-7. DOI: 10.1038/nmeth.1176. View

Yildiz A, Tomishige M, Vale R, Selvin P . Kinesin walks hand-over-hand. Science. 2003; 303(5658):676-8. DOI: 10.1126/science.1093753. View

Hell S . Microscopy and its focal switch. Nat Methods. 2009; 6(1):24-32. DOI: 10.1038/nmeth.1291. View

Nienhaus G, Nienhaus K, Holzle A, Ivanchenko S, Renzi F, Oswald F . Photoconvertible fluorescent protein EosFP: biophysical properties and cell biology applications. Photochem Photobiol. 2006; 82(2):351-8. DOI: 10.1562/2005-05-19-RA-533. View

Michalet X, Pinaud F, Bentolila L, Tsay J, Doose S, Li J . Quantum dots for live cells, in vivo imaging, and diagnostics. Science. 2005; 307(5709):538-44. PMC: 1201471. DOI: 10.1126/science.1104274. View

Fernandez-Suarez M, Ting A . Fluorescent probes for super-resolution imaging in living cells. Nat Rev Mol Cell Biol. 2008; 9(12):929-43. DOI: 10.1038/nrm2531. View

Shaner N, Patterson G, Davidson M . Advances in fluorescent protein technology. J Cell Sci. 2007; 120(Pt 24):4247-60. DOI: 10.1242/jcs.005801. View

10.

Folling J, Bossi M, Bock H, Medda R, Wurm C, Hein B . Fluorescence nanoscopy by ground-state depletion and single-molecule return. Nat Methods. 2008; 5(11):943-5. DOI: 10.1038/nmeth.1257. View

11.

Donnert G, Eggeling C, Hell S . Major signal increase in fluorescence microscopy through dark-state relaxation. Nat Methods. 2006; 4(1):81-6. DOI: 10.1038/nmeth986. View

12.

Heilemann M, Herten D, Heintzmann R, Cremer C, Muller C, Tinnefeld P . High-resolution colocalization of single dye molecules by fluorescence lifetime imaging microscopy. Anal Chem. 2002; 74(14):3511-7. DOI: 10.1021/ac025576g. View

13.

Lavis L, Raines R . Bright ideas for chemical biology. ACS Chem Biol. 2008; 3(3):142-55. PMC: 2802578. DOI: 10.1021/cb700248m. View

14.

Chen I, Ting A . Site-specific labeling of proteins with small molecules in live cells. Curr Opin Biotechnol. 2005; 16(1):35-40. DOI: 10.1016/j.copbio.2004.12.003. View

15.

Moerner , Kador . Optical detection and spectroscopy of single molecules in a solid. Phys Rev Lett. 1989; 62(21):2535-2538. DOI: 10.1103/PhysRevLett.62.2535. View

16.

Juette M, Gould T, Lessard M, Mlodzianoski M, Nagpure B, Bennett B . Three-dimensional sub-100 nm resolution fluorescence microscopy of thick samples. Nat Methods. 2008; 5(6):527-9. DOI: 10.1038/nmeth.1211. View

17.

Bates M, Blosser T, Zhuang X . Short-range spectroscopic ruler based on a single-molecule optical switch. Phys Rev Lett. 2005; 94(10):108101. PMC: 2652517. DOI: 10.1103/PhysRevLett.94.108101. View

18.

Thompson R, Larson D, Webb W . Precise nanometer localization analysis for individual fluorescent probes. Biophys J. 2002; 82(5):2775-83. PMC: 1302065. DOI: 10.1016/S0006-3495(02)75618-X. View

19.

Betzig E, Chichester R . Single molecules observed by near-field scanning optical microscopy. Science. 1993; 262(5138):1422-5. DOI: 10.1126/science.262.5138.1422. View

20.

Ando R, Mizuno H, Miyawaki A . Regulated fast nucleocytoplasmic shuttling observed by reversible protein highlighting. Science. 2004; 306(5700):1370-3. DOI: 10.1126/science.1102506. View