Nanobodies: Natural Single-Domain Antibodies

Annual Review of Biochemistry, Journal Year: 2013, Volume and Issue: 82(1), P. 775 - 797

Published: March 15, 2013

Sera of camelids contain both conventional heterotetrameric antibodies and unique functional heavy (H)-chain (HCAbs). The H chain these homodimeric consists one antigen-binding domain, the VHH, two constant domains. HCAbs fail to incorporate light (L) chains owing deletion first domain a reshaped surface at VHH side, which normally associates with L in antibodies. genetic elements composing have been identified, but vivo generation from their dedicated genes into antigen-specific affinity-matured bona fide remains largely underinvestigated. However, facile identification VHHs beneficial biochemical economic properties (size, affinity, specificity, stability, production cost) supported by multiple crystal structures encouraged antibody engineering single-domain for use as research tool biotechnology medicine.

Language: Английский

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Multiplexed 3D cellular super-resolution imaging with DNA-PAINT and Exchange-PAINT

Nature Methods, Journal Year: 2014, Volume and Issue: 11(3), P. 313 - 318

Published: Feb. 2, 2014

Language: Английский

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Super-resolution microscopy demystified

Nature Cell Biology, Journal Year: 2018, Volume and Issue: 21(1), P. 72 - 84

Published: Dec. 17, 2018

Language: Английский

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Super-resolution microscopy with DNA-PAINT

Nature Protocols, Journal Year: 2017, Volume and Issue: 12(6), P. 1198 - 1228

Published: May 18, 2017

Language: Английский

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Fluorescence nanoscopy in cell biology

Nature Reviews Molecular Cell Biology, Journal Year: 2017, Volume and Issue: 18(11), P. 685 - 701

Published: Sept. 6, 2017

Language: Английский

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Single-molecule localization microscopy

Nature Reviews Methods Primers, Journal Year: 2021, Volume and Issue: 1(1)

Published: June 3, 2021

Single-molecule localization microscopy (SMLM) describes a family of powerful imaging techniques that dramatically improve spatial resolution over standard, diffraction-limited and can image biological structures at the molecular scale. In SMLM, individual fluorescent molecules are computationally localized from sequences localizations used to generate super-resolution or time course images, define trajectories. this Primer, we introduce basic principles SMLM before describing main experimental considerations when performing including labelling, sample preparation, hardware requirements acquisition in fixed live cells. We then explain how low-resolution processed reconstruct images and/or extract quantitative information, highlight selection discoveries enabled by closely related methods. discuss some limitations potential artefacts as well ways alleviate them. Finally, present an outlook on advanced promising new developments fast-evolving field SMLM. hope Primer will be useful reference for both newcomers practitioners

Language: Английский

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Chromatin Fibers Are Formed by Heterogeneous Groups of Nucleosomes In Vivo

Cell, Journal Year: 2015, Volume and Issue: 160(6), P. 1145 - 1158

Published: March 1, 2015

Language: Английский

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Precisely and accurately localizing single emitters in fluorescence microscopy

Nature Methods, Journal Year: 2014, Volume and Issue: 11(3), P. 253 - 266

Published: Feb. 27, 2014

Language: Английский

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A robust pipeline for rapid production of versatile nanobody repertoires

Nature Methods, Journal Year: 2014, Volume and Issue: 11(12), P. 1253 - 1260

Published: Nov. 2, 2014

Language: Английский

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Three-Dimensional Localization of Single Molecules for Super-Resolution Imaging and Single-Particle Tracking

Chemical Reviews, Journal Year: 2017, Volume and Issue: 117(11), P. 7244 - 7275

Published: Feb. 2, 2017

Single-molecule super-resolution fluorescence microscopy and single-particle tracking are two imaging modalities that illuminate the properties of cells materials on spatial scales down to tens nanometers or with dynamical information about nanoscale particle motion in millisecond range, respectively. These methods generally use wide-field microscopes two-dimensional camera detectors localize molecules much higher precision than diffraction limit. Given limited total photons available from each single-molecule label, both require careful mathematical analysis image processing. Much more can be obtained system under study by extending three-dimensional (3D) localization: without this capability, visualization structures motions axial direction easily missed confused, compromising scientific understanding. A variety for obtaining 3D images have been devised, their own strengths weaknesses. include multiple focal planes, point-spread-function engineering, interferometric detection. may compared based ability provide accurate precise position emitters photons. To successfully apply further develop these methods, it is essential consider many practical concerns, including effects optical aberrations, field dependence system, fluorophore labeling density, registration between different color channels. Selected examples described illustration a biological contexts demonstrating power localization understanding complex systems.

Language: Английский

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