Warren W S Invited Review Article Pump-probe Microscopy Rev Sci Instrum

Basics

Unmarried-Molecule Probes of Nanoenvironments in Condensed Affair, Explorations in Solids or Cells Using a Nanometer-Sized Probe, or Probing Local Environments Using Single-Molecule Spectroscopy and Microscopy.

Effigy 1: Imaging Single Molecules of Pentacene in a p-Terphenyl Crystal

Near chemic experiments in condensed matter mensurate the boilerplate beliefs of a huge number, N, of molecules, where N may range from millions to billions to Avogadro's Number. At the same time, nigh theoretical models are intended to depict the behavior of a single molecule interacting with its surroundings, and averaging over the number of molecules N is normally required to compute an observable. Using precision laser spectroscopic techniques, we have been detecting and probing the detailed backdrop of individual impurity molecules hidden deep inside a solid, a protein, or even in a liquid, i.due east., the ultimate limit of N=1. This was first done in the Moerner Lab in 1989, and has since expanded dramatically to include many groups around the world. A key reason for doing this is to explore heterogeneity that is commonly obscured past ensemble averaging.

Studying i individual molecule in a solid means we are working with an extremely small number of moles of material. You might be aware that the international standards organization, IUPAC, has divers several new prefixes: zepto- for 1E-21, and yocto- for 1E-24. Thus 1 molecule is equivalent to 1.66 yoctomoles. But we call up this is unwieldy. Thus we define a new prefix guaca- so that (with apologies to Prof. Avogadro)

1 guacamole = 1 / ( Avocado'south Number) of moles.

More seriously, information technology is worth recalling that each molecule nosotros are probing is only 1 or 2 nanometers in size. This means that when we use a laser to select 1 probe molecule, we can sense details of the firsthand local environment of a truly nanoscopic probe.

To achieve this farthermost reduction of the concentration and reach the single-molecule level, nosotros apply either (a) low-temperature loftier-resolution light amplification by stimulated emission of radiation spectroscopy to select private molecules past their resonance frequencies, or (b) nearly field optical excitation to pump sample volumes much smaller than the diffraction limit, or (c) extremely depression concentrations and diffraction-limited confocal or far-field microscopy. By studying a big number of individual molecules ane at a time, we are able not only to observe how the usual ensemble average beliefs is formed, but besides to see unexpected, surprising behavior normally hidden by the usual ensemble averaging.

The phenomena under study in the early on work at low temperatures included spontaneous changes in resonant frequency caused past host matrix dynamics (spectral diffusion) as well every bit light-induced resonant frequency changes, akin in principle to optical storage on a single molecule level. Since a very narrow optical transition is extremely sensitive to perturbations, extremely small changes in the local surroundings were probed by the application of electrical, strain, or magnetic fields. By dispersing the emitted low-cal, even the vibrational mode spectrum of a single molecule was measured! By measuring correlations in the emitted photon stream, fast dynamics including ecology fluctuations, or the purely breakthrough-mechanical behavior termed photon antibunching may be probed.

At room temperature, a huge variety of additional physical and chemical effects can be studied spanning multiple fields from biology to materials scientific discipline. In biomolecules, we observe fascinating differences in behavior of single molecules due to conformational states, local environments, or enzymatic cycle, all of which are obscured in big Northward experiments. Unmarried labeled biomolecules tin can be observed even in living cells, where now the behaviors of private nanomachines can exist addressed. Since the unmarried fluorescent dye characterization acts as a nanometer-sized light source, today single-molecule imaging has provided a pathway to surpass the optical diffraction limit.

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Nobel Lecture, Autobiography, and Written Nobel Lecture

Nobel Lecture Page with full details: Video, written article, and slides: "William East. Moerner - Nobel Lecture: Unmarried-Molecule Spectroscopy, Imaging, and Photocontrol: Foundations for Super-Resolution Microscopy".Nobelprize.org. Nobel Media AB 2014. Spider web. 19 Nov 2015. <http://www.nobelprize.org/nobel_prizes/chemistry/laureates/2014/moerner-lecture.html>

Nobel Lecture Article with a few typo corrections: Angewandte Chemie and Reviews of Modern Physics

Nobel Autobiography (as of January 2015): "William Eastward. Moerner - Biographical".Nobelprize.org. Nobel Media AB 2014. Spider web. 19 November 2015. <http://www.nobelprize.org/nobel_prizes/chemistry/laureates/2014/moerner-bio.html>

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Selected Bibliography and Reviews (chronological)

• "Optical Detection and Spectroscopy of Single Molecules in a Solid," by W. Due east. Moerner and L. Kador, Phys. Rev. Lett. 62, 2535 (1989). This is the first report of single-molecule detection and spectroscopy in condensed phases.
• "Fluorescence Spectroscropy and Spectral Diffusion of Single Impurity Molecules in a Crystal," past W.P. Ambrose and W. E. Moerner, Nature 349, 225 (1991).
• "Spectroscopy of Single Impurity Molecules in Solids," past W. E. Moerner and Th. Basche', Angew. Chem. 105, 537 (1993); Angew. Chem. Int. Ed. Engl. 32, 457 (1993).
• "Examining Nanoenvironments in Solids on the Scale of a Single, Isolated Impurity Molecule," by West. Due east. Moerner, Science 265, 46 (1994).
• "High-Resolution Optical Spectroscopy of Unmarried Molecules in Solids, past Westward. East. Moerner, in "Single Molecules and Atoms," Special Outcome of Accounts of Chemical Research, December 1996.
• "Fundamentals of Single-Molecule Spectroscopy in Solids," Chapter 1 of Single Molecule Optical Detection, Imaging, and Spectroscopy, T. Basche, W. E. Moerner, Thousand. Orrit, and U. P. Wild, eds. (Verlag Chemie, Munich, 1997).
• West. Eastward. Moerner, "Those Blinking Single Molecules," Science 277, 1059 (1997).
• W. Eastward. Moerner and M. Orrit, "Illuminating Single Molecules in Condensed Matter," Science 283, 1670-1676 (1999).
• B. Lounis and W. E. Moerner, "Single Photons on Demand from a Single Molecule at Room Temperature," Nature 407, 491-493 (2000).
• H. Sosa, Eastward. J. Chiliad. Peterman, W. E. Moerner, and L. S. B. Goldstein, "ADP-Induced Rocking of the Kinesin Motor Domain Revealed by Single-Molecule Fluorescence Polarization Microscopy," Nature Structural Biology 8, 540-544 (2001).
• W. Due east. Moerner, "13 Years of Single-Molecule Spectroscopy in Physical Chemical science and Biophysics," in Single-Molecule Spectroscopy: Nobel Conference Lectures, R. Rigler, M. Orrit, Thursday. Basche, Editors, Springer Serial in Chemical Physics, Volume 67 (Springer-Verlag, Heidelberg, 2001), pp. 32-61.
• W. E. Moerner, "A Dozen Years of Single-Molecule Spectroscopy in Physics, Chemistry, and Biophysics," J. Phys. Chem. B 106, 910-927 (2002).
• West. E. Moerner, "Single-Molecule Optical Spectroscopy of Autofluorescent Proteins," J. Chem. Phys. 117, 10925 (2002).
• Westward. E. Moerner and D. P. Fromm, "Methods of Single-Molecule Fluorescence Spectroscopy and Microscopy," Rev. Sci. Instrum. 74, 3597-3619 (2003).
• East. J. G. Peterman, H. Sosa, and West. East. Moerner, "Unmarried-Molecule Fluorescence Spectroscopy and Microscopy of Biomolecular Motors," invited review, Ann. Rev. Phys. Chem. 55, 79-96 (2004).
• P. J. Schuck, D. P. Fromm, A. Sundaramurthy, M. Due south. Kino, and W. East. Moerner, "Improving the Mismatch Betwixt Light and Nanoscale Objects with Gold Bowtie Nanoantennas," Phys. Rev. Lett. 94, 017402 (2005).
• Chiliad. A. Willets, S. Y. Nishimura, P. J. Schuck, R. J. Twieg, and West. East. Moerner, "Nonlinear Optical Chromophores every bit Nanoscale Emitters for Single-Molecule Spectroscopy," invited review, Accounts Chem. Res. 38, 549-556 (2005)
• S. Y. Kim, Z. Gitai, A. Kinkhabwala, Fifty. Shapiro, and W. Due east. Moerner, "Unmarried Molecules of the Bacterial Actin MreB Undergo Directed Treadmilling Motion in Caulobacter crescentus," Proc. Nat. Acad. Sci. (U.s.) 103, 10929-10934 (2006).
• A. E. Cohen and W. East. Moerner, "Suppressing Brownian Motion of Individual Biomolecules in Solution," Proc. Nat. Acad. Sci. (USA) 103, 4362-4365 (2006).
• W. Due east. Moerner, "New Directions in Unmarried-Molecule Imaging and Analysis," Invited Perspective, Proc. Nat. Acad. Sci. (USA) 104, 12596-12602 (2007).
• J. South. Biteen, Yard. A. Thompson, N. K. Tselentis, G. R.Bowman, L. Shapiro, W. Eastward. Moerner, "Superresolution Imaging in Live Caulobacter Crescentus Cells Using Photoswitchable EYFP," Nature Meth. 5, 947-949 (2008).
• W. East. Moerner, "Single-Molecule Optical Spectroscopy and Imaging: From Early Steps to Recent Advances," in Single Molecule Spectroscopy in Chemistry, Physics and Biology: Nobel Symposium 138, Springer Series in Chemical Physics Vol. 96, A. Gräslund, R. Rigler, J. Widengren, Eds. ( Springer-Verlag, Berlin, 2009), pp. 25-60.
• A. Kinkhabwala, Z. Yu, Southward. Fan, Y. Avlasevich, Chiliad. Müllen, and W. Eastward. Moerner, "Large Single-Molecule Fluorescence Enhancements Produced past a Bowtie Nanoantenna," Nature Photonics 3, 654-657 (2009).
• South. J. Lord, H.-Fifty. D. Lee, and W. E. Moerner, "Single-Molecule Spectroscopy and Imaging of Biomolecules in Living Cells," Anal. Chem. 82, 2192-2203 (2010) .
• K. A. Thompson, J. S. Biteen, Due south. J. Lord, Northward. R. Conley, and W. E. Moerner, "Molecules and Methods for Super-Resolution Imaging," in Methods in Enzymology, Volume 475, Nils G. Walter, Editor (Elsevier, New York, 2010), Chapter ii, pp. 27-59. DOI
• Michael A. Thompson, Matthew D. Lew, and W. East. Moerner, "Extending Microscopic Resolution with Single-Molecule Imaging and Active Control," Annual Reviews of Biophysics 41, 321-342 (2012). DOI
• Matthew D. Lew, Steven F. Lee, Michael A. Thompson, Hsiao-lu D. Lee, and W. Due east. Moerner, "Unmarried-Molecule Photocontrol and Nanoscopy,"  in Far-Field Optical Nanoscopy, P. Tinnefeld, C. Eggeling, and Due south. Due west. Hell, Eds., Springer Series on Fluorescence (Springer, Berlin, Heidelberg, 2012). DOI
• Quan Wang, Randall H. Goldsmith, Yan Jiang, Samuel D. Bockenhauer, and W.Due east. Moerner, "Probing single biomolecules in solution using the Anti-Brownian ELectrokinetic (ABEL) trap," Acc. Chem. Res. 45, 1955-1964 (Paul Barbara Special Issue) (2012). DOI
• W. E. Moerner, "Microscopy beyond the diffraction limit using actively controlled single molecules," J. Microsc. 246, 213-220 (2012). DOI
• Steffen J. Sahl and W. Due east. Moerner, "Super-resolution Fluorescence Imaging with Single Molecules," Curr. Opin. Struct. Biol. 23, 778-787 (2013). DOI
• Andreas Gahlmann and W. Due east. Moerner, "Exploring bacterial cell biology with single-molecule tracking and super-resolution imaging," Nature Reviews Microbiology 12, 9-22 (2014), published online Dec xvi, 2013. DOI
• Mikael P. Backlund, Matthew D. Lew, Adam Southward. Backer, Steffen J. Sahl, and W. E. Moerner, "The role of molecular dipole orientation in single-molecule fluorescence microscopy and implications for super-resolution imaging," Minireview, ChemPhysChem xv, 587-599 (2014), published online Dec 30, 2013. DOI
• West. E. Moerner, Yoav Shechtman, and Quan Wang, "Unmarried-molecule spectroscopy and imaging over the decades," Faraday Hash out. 20, one-28 (2015) (DOI:10.1039/c5fd00149h, published online November 30, 2015). DOI
• Alex von Diezmann, Yoav Shechtman, and W. E. Moerner, "Three-Dimensional Localization of Unmarried Molecules for Super-Resolution Imaging and Single-Particle Tracking," Chem. Revs. Special Issue on Super-Resolution and Single-Molecule Imaging (in press, DOI: 10.1021/acs.chemrev.6b00629, published online 2 February 2017). DOI
• Leonhard Möckl and W. E. Moerner, "Super-resolution microscopy with unmarried molecules in biology and beyond – essentials, current trends, and hereafter challenges," Perspective Article, J. Am. Chem. Soc. 142, 17828-17844 (2020) (DOI: 10.1021/JACS.0c08178, published online 9 October 2020).

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Source: https://web.stanford.edu/group/moerner/sms_gen.html

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