title: Nanoscale devices for linkerless long-term single-molecule observation.
abstract: Total internal reflection fluorescence microscopy (TIRFM) can offer favorably high signal-to-noise observation of biological mechanisms. TIRFM can be used routinely to observe even single fluorescent molecules for a long duration (several seconds) at millisecond time resolution. However, to keep the investigated sample in the evanescent field, chemical surface immobilization techniques typically need to be implemented. In this review, we describe some of the recently developed novel nanodevices that overcome this limitation enabling long-term observation of free single molecules and outline their biological applications. The working concept of many devices is compatible with high-throughput strategies, which will further help to establish unbiased single molecule observation as a routine tool in biology to study the molecular underpinnings of even the most complex biological mechanisms.
authors.affiliation: Structural and Computational Biology Unit and Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, 69117 Heidelberg, Germany.
authors.forename: Niccolò
authors.initials: N
authors.lastname: Banterle
authors.affiliation: Structural and Computational Biology Unit and Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, 69117 Heidelberg, Germany. Electronic address: lemke@embl.de.
authors.forename: Edward A
authors.initials: EA
authors.lastname: Lemke
mesh_terms: D000818:Animals; D002468:Cell Physiological Phenomena; D003584:Cytological Techniques / Q000379:methods*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D036103:Nanotechnology / Q000295:instrumentation*; D059010:Single-Cell Analysis / Q000295:instrumentation* / Q000379:methods*